Janet H Leung*1, Jessica Lasky-Su1, Jonathan S Williams2 and Gordon H Williams1
1Brigham and Women's Hospital, Harvard Medical School, Boston, MA, 2Brigham & Women's Hospital, Harvard Medical School, Boston, MA


Previous studies have documented that genetic variants in the β2 adrenergic receptor (β2AR) are associated with salt-sensitive hypertension in Caucasians. [1] Similarly, variance in lysine specific demethylase-1 (LSD-1) is associated with salt-sensitivity in hypertensive African-Americans. [2] From animal studies, these two genes appear to modify aldosterone secretion in response to changes in sodium intake. However, in vivo in humans, genes do not function in isolation, and interactions may explain why gene variants that are associated with large changes in systolic blood pressure (ΔSBP) in animals are associated with much smaller ΔSBP in humans.  Thus, we tested the hypothesis that there is a gene-gene interaction between β2AR and LSD-1 with salt-sensitivity of blood pressure in humans. We additionally evaluated whether the interaction is ethnicity-specific.

We utilized the Hypertension Pathotypes cohort, a multinational, multi-decade study of the genetic underpinnings of hypertension in both hypertensives and normotensives. LSD-1 and β2AR are represented by tagging SNPs rs587168 and rs1042713, respectively. All participants consumed a low sodium (<20 mmol/day) then a liberal sodium diet (> 200 mmol/day) for 7 days. The outcome of interest was the change in ΔSBP between the two diets.  Statistical comparison of ΔSBP were performed with an interaction term for each pair of SNPs coded as “risk” or “non-risk” based on prior published analyses, and then with models adjusting for age, gender, BMI, and known diagnosis of hypertension individually and in a full model. The analysis was in Caucasian (n=468) and African–descent (n=73) subjects who had phenotype and genotype data.

In those of African-descent with and without hypertension, β2AR gene variance has no independent effect on ΔSBP (p = 0.8). LSD-1 has a minimally significant effect on ΔSBP (beta 3.7, p = 0.123 in an unadjusted model). However, the interaction between the two SNPs was highly significant (beta_interaction -25.1, p <0.001), and remained so even after adjusting for age, gender, BMI, and diagnosis of hypertension (beta -24.1; p <0.001). Of note, of these classic predictors, only female gender was a significant confounder. Unadjusted ΔSBP with no risk alleles was 6.0 mm Hg; 19.3 and 20.8 for having a single risk genotype (β2AR risk and LSD-1, respectively); and 8.8 mm Hg for having both. Similar analysis for interaction in the Caucasian cohort was not significant, with a p-value of 0.3.

Thus, LSD-1 and β2 adrenergic receptor risk genotypes interact in a not only a non-additive, but potentially protective, manner in populations of African-descent but not Caucasians. That the presence of two risk alleles was associated with less salt-sensitivity than having either risk allele alone may explain in part blood pressure heterogeneity within a population. Future work will explore the interaction of genes via a network model.


Disclosure: GHW: Ad Hoc Consultant, Daiichi Sankyo, Ad Hoc Consultant, Relypsa, Ad Hoc Consultant, Pfizer Global R&D, Ad Hoc Consultant, Tanabe. Nothing to Disclose: JHL, JL, JSW

PP02-2 27183 2.0000 FRI 575 A Having Two Risk Genotypes for Salt-Sensitive Hypertension Associates with Lower Change in Systolic Blood Pressure Than Either Alone 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 11:45:00 AM PP02 7663 11:30:00 AM Renin-Angiotensin-Aldosterone System - Bench to Bedside Poster Preview

Takumi Kitamoto*1, Sachiko Suematsu2, Yoko Matsuzawa2, Jun Saito2, Masao Omura2 and Tetsuo Nishikawa2
1Graduate School of Medicine, Chiba University, Chiba, Japan, 2Yokohama Rosai Hosp, Yokohama, Japan


Prevalence of KCNJ5 gene mutations is almost 70% among Japanese patients with APAs. APA is not always rare among hypertensive patients, while incomplete remission of hypertension (HTN) is sometimes observed after surgery. Thus, we attempted to investigate whether or not KCNJ5 gene mutations affect an improvement in HTN and its complications in those patients after unilateral adrenalectomy.

We enrolled 104 APA patients and sequenced KCNJ5, ATPase and CACNA1D cDNA in APA-tissues to compare the clinical characteristics between KCNJ5 mutated and wild type group. We also retrospectively analyzed the patients before and 1 year after surgery

Somatic KCNJ5 gene mutation was detected in 75 APAs and 24 didn’t show any mutations. The other APAs were detected somatic mutations as follows; ATP1A1 mutation in 1 APA, ATP2B3 in 2 APAs, CACNA1D in 2 APAs. KCNJ5 mutated APA patients showed significantly younger age, severer hypokalemia, higher plasma aldosterone concentration and lower renin activity than those with wild type APAs, although BMI, duration of HTN, blood pressure, number of anti-hypertensive drugs and the prevalence of non-dipper HTN were similar between each group. HTN was post-operatively improved in all patients, although complete remission (CR) was much more achieved in the mutated group (65%) than in the wild-type group (35%). In KCNJ5 mutated group, the patients with CR of HTN showed significantly younger age, shorter duration of HTN, lower BMI, smaller number of anti-hypertensive drugs, higher eGFR and milder vascular complications than those with partial remission (PR) before surgery. Aldosterone production level and frequency of non-dipper HTN were not different between the patients with CR and PR. 1 year after surgery, vascular complications, such as left ventricular hypertrophy, arterial stiffness and inner carotid artery thickness, were much more improved in KCNJ5 mutated group than in the wild-type group.

The present study clearly demonstrated that early surgical treatment is quite effective for achieving CR, especially in KCNJ5 mutated APA-patients, since CR was more frequently observed in the KCNJ5 mutated group than in the wild type group regardless of similar incidences of non-dipper HTN in both groups. Moreover, unilateral adrenalectomy in KCNJ5 mutated patients could make much more beneficial effect on improving vascular complications than in the wild-type APA-patients. Thus, we should precisely diagnose younger APA-patients with KCNJ5 gene mutations for improving blood pressure and vascular complications.


Nothing to Disclose: TK, SS, YM, JS, MO, TN

PP02-3 25224 3.0000 FRI 573 A Detection of KCNJ5 Mutated Aldosterone-Producing Adenoma (APA) Can Predict Remission of Hypertension and Its Complications 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 11:45:00 AM PP02 7663 11:30:00 AM Renin-Angiotensin-Aldosterone System - Bench to Bedside Poster Preview

Hiroki Shimada*1, Ryo Ito1, Ikuko Sato1, Emiko Sato2, Atsushi Yokoyama1 and Akira Sugawara1
1Tohoku Univ Grad School of Med, Sendai, Japan, 2Tohoku Univ Grad Sch of Pharmaceutical Sciences, Sendai



It is well known that obese patients are highly complicated with hypertension. Recently, “undetermined” adipocyte-derived factor(s) have been recognized as one of the etiologies of obesity-related hypertension independent of angiotensin (A) II. In the present study, we aim to identify adipocyte-derived factor(s) that stimulate aldosterone synthase gene (CYP11B2) expression/aldosterone secretion that may induce obesity-related hypertension.


We first differentiated mouse fibroblast 3T3-L1 cells into adipocytes, and collected their supernatants. The supernatants were then incubated with human adrenocortical carcinoma-derived H295R cells, and their ability to stimulate CYP11B2 mRNA expression was determined by qRT-PCR using RNA extracted from H295R cells. Thereafter, the supernatants were fractionated by ultrafiltration to obtain active fractions that stimulate CYP11B2 expression, and proteins included in the fractions were analyzed by LC-MS/MS.


The supernatants obtained from 3T3-L1 adipocytes were shown to stimulate CYP11B2 expression, whose activity was abolished by heating at 96°C for 5 min. AII could not be observed in the supernatants when determined by LC-MS/MS. Fractionation by molecular weight using ultrafiltration demonstrated that the ability to stimulate CYP11B2 expression was detected in fractions between 50 and 100 kDa. The proteins included in the fractions were analyzed by LC-MS/MS, and several candidates were obtained. We are now trying to identify adipocyte-derived factor(s) that can stimulate CYP11B2 expression from them.


We here have demonstrated the existence of “undetermined” adipocyte-derived factor(s) that have the ability of stimulate CYP11B2 expression. The factor(s) were heat sensitive, different from AII, and their molecular weight was between 50 and 100 kDa. The factor(s) may therefore be one of the causes of hypertension and arteriosclerosis through the increase of aldosterone secretion in obese patients. The structural and functional elucidation of the factor(s) will give us clues for the future innovation of drugs against obesity-related hypertension.


Nothing to Disclose: HS, RI, IS, ES, AY, AS

PP02-4 25660 4.0000 FRI 574 A A Novel Approach for the Identification of Undetermined Adipocyte-Derived Factor(s) That Stimulate Aldosterone Synthase Gene (CYP11B2) Expression 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 11:45:00 AM PP02 7663 11:30:00 AM Renin-Angiotensin-Aldosterone System - Bench to Bedside Poster Preview

Govindan Dayanithi*1, Stepan Kortus2, Oksana Forostyak3, Eva Sykova2 and Alexei Verkhratsky4
1Institute of Experimental Medicine-AS CR & INSERM 1198-France, Prague, Czech Republic, 2Institute of Experimental Medicine-AS CR, Prague, Czech Republic, 32nd Faculty of Medicine, Charles University in Prague, Prague, Czech Republic, 4University of Manchester, Manchester, United Kingdom


The magnocellular vasopressin (AVP) and oxytocin (OT) neurons exhibit specific electrical behavior, synthesize AVP and OT peptides, and secrete them into the neurohypophysial system in response to various physiological stimulants. The electrical activities of these neurons are regulated by AVP and OT, either released from soma and dendrites or applied to isolated supraoptic nucleus (SON) neurons or to slice preparations. In these neurons, both AVP and OT bind to specific autoreceptors which induce distinct Ca2+ signals and regulate cellular events (1). Using Fura-2 fast fluorescence microspectrofluorimetry or video imaging, we measured [Ca2+]i oscillations in single freshly dissociated neurons from the adult (12-16 weeks-old) non-transgenic and transgenic Wistar rats (AVP-eGFP) (2). In transgenic rats, the neurons were identified under a fluorescence microscope with GFP filters; in non-transgenic rats, the neurons were identified by their specific [Ca2+]i responses to AVP (3). More than 80% of AVP-eGFP neurons exhibited these spontaneous [Ca2+]i oscillations. Only a few (less than 20%) of non-AVP-eGFP neurons were oscillatory. In AVP-eGFP or AVP-sensitive neurons, AVP modulated the oscillations in a manner that depends on the initial state of the neuron. In silent neurons, AVP triggered [Ca2+]i oscillations whereas it inhibited oscillations when the neurons were already oscillating. This behavior could be correlated with in vivo observations: excitatory, inhibitory, or no effect, depending on the initial firing pattern. In silent neurons, V1a- and V2-type vasopressin receptor agonists (F180: 100nM; and dDAVP: 100 nM, respectively) triggered the oscillations. The oscillations were not affected by the AVP-V1-type receptor antagonist, ([deamino-Pen1, O-Me-Tyr2, Arg8]-vasopressin) suggesting that the oscillations are not mediated by AVP receptors. However, the oscillations were disrupted following exposure to hypertonic or hypotonic solutions (325, 275 mOsm/kg, respectively) and were altered in neurons obtained from rats that had been dehydrated for 3 to 5 days. Furthermore, our results revealed that [Ca2+]i oscillations are driven by several components localized at the plasma membrane and that the mitochondrial oxidative activity is required to sustain the oscillations. These results unveil for the first time the idiosyncrasies of Ca2+ signaling in AVP neurons that contribute to maintain their remarkable intrinsic in vivo physiological properties in isolated conditions.


Nothing to Disclose: GD, SK, OF, ES, AV

PP05-1 24021 1.0000 FRI 476 A Calcium Oscillations in the Isolated Vasopressin Neurons of the Rat Supraoptic Nucleus 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 11:45:00 AM PP05 7674 11:30:00 AM Neuroendocrinology Poster Preview

Gislaine Almeida-Pereira1, Ricardo Coletti2, Tatiane Vilhena-Franco3, Susana Quirós Cognuck3, Hellen Veiga Silva3, Lucila Elias4 and Jose Antunes-Rodrigues*4
1Ribeirao Preto Medical School, University of Sao Paulo, Ribeirao Preto, Brazil, 2Universidade de São Paulo, Ribeirão Preto, Brazil, 3School of Medicine of Ribeirao Preto - University of Sao Paulo, Ribeirao Preto, Brazil, 4School of Medicine of Ribeirao Preto, University of Sao Paulo, Ribeirao Preto, Brazil


Introduction: The angiotensin II (ANGII) plays an important role in the control of body fluids and estradiol (E2) modulates several actions of ANGII in the brain (Fitzsimons, 1998). However, the cellular mechanisms of the interaction between E2 and ANGII and its physiological role in the control of body fluids remain unclear. It is known that activation of the PKC signaling pathway is associated with the activation of p38MAPK or ERK1/2 in several systems, but some studies have demonstrated that stimulation of AT1 receptor induces activation of ERK1/2 and PKC in an independent manner (Daniels et al. 2007). In addition, estrogen receptor (ER) is known for its classic genomic actions, but recent studies have shown that estrogens also activate nongenomic cellular signaling events, such as PKC and MAPK mediated pathways (McEwen, 2001). Therefore, this study aimed to investigate the role of estradiol, PKC, p38MAPK and ERK1/2 signaling pathways on the ANGII-induced oxytocin (OT) and vasopressin (AVP) secretion in OVX female rats.

Methods: Wistar rats (~250g) were submitted to ovariectomy (OVX) and on the following day they were treated with estradiol cypionate (10µg/rat, sc, OVX+E2) or vehicle (corn oil, 0.1mL/rat, sc) for eight days. On the eighth day, the rats received an icv (lateral ventricle) injection of ERK1/2 inhibitor (U0126, 1mM/2µL/rat) or p38MAPK inhibitor (SB203580, 50µM/2µL/rat) or PKC inhibitor (Chelerythrine, 1mM/ 2µL/rat) or vehicle (DMSO 5%, 2µL/rat) and 20 min after they were injected with angiotensin II (ANGII, 25ng/2μL/rat) or vehicle (0.9% saline, 2μL/rat). After five min of ANGII injection the animals were decapitated for blood collection for OT and AVP analysis by specific radioimmunoassays. Data were analyzed using ANOVA three-way, followed by Newman-Keuls post-test and the level of significance was set at 5%.

Results: The pretreatment with estradiol attenuated both OT (ANGII x E2 interaction: F=11.0, df=1, p<0.05, n=9) and AVP secretion (ANGII x E2 interaction: F=8.2, df=1, p<0.05, n=9) induced by ANGII in OVX rats. PKC and p38MAPK inhibition blocked OT secretion (ANGII x inhibitor interaction: F=5.2, df=1, p<0.05, n=17; F=7.4, df=1, p<0.05, n=13, respectively). On the other hand, ERK1/2 inhibition attenuated AVP (ANGII x inhibitor interaction: F=6.6, df=1, p<0.05, n=13) induced by ANGII in OVX rats. PKC, p38MAPK and ERK1/2 inhibitors did not affect the ANGII-induced OT and AVP secretion in OVX+E2 rats.

Conclusions: The present results show for the first time the physiological relevance of PKC, p38MAPK and ERK1/2 signaling pathways in the neurohypophysial hormone release induced by ANGII in female rats. These data suggest that ANGII evokes dual signaling pathways, ERK1/2 for AVP secretion and PKC/p38MAPK for OT secretion.



Nothing to Disclose: GA, RC, TV, SQC, HVS, LE, JA

PP05-2 26840 2.0000 FRI 477 A PKC/p38MAPK and ERK1/2 Dual Signaling Pathways Mediate the Angiotensin Effects on Neurohypophysial Hormone Secretion in Female Rats 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 11:45:00 AM PP05 7674 11:30:00 AM Neuroendocrinology Poster Preview

Michelle N. Bedenbaugh*, Justin A. Lopez, Richard B. McCosh, Robert L. Goodman and Stanley M. Hileman
West Virginia University School of Medicine, Morgantown, WV


Increased secretion of GnRH into the portal vasculature is critical for the initiation of puberty.  Estradiol exerts negative feedback on GnRH neurons and suppresses secretion during the prepubertal period.  However, GnRH neurons do not express estrogen receptor alpha (ERα).  Therefore, intermediates that express ERα must exist to communicate estradiol negative feedback to GnRH. Nitric oxide (NO) influences LH secretion, and inhibition of nitric oxide synthase delays puberty in rats.  However, little is known about the relationship between NO and ERα expression in sheep.  Therefore, this study aimed to characterize ERα expression in hypothalamic populations of neuronal nitric oxide synthase (nNOS) neurons in the presence or absence of estradiol negative feedback. Hypothalamic tissue from prepubertal ewes that were either ovariectomized (OVX; n=6) or ovariectomized and implanted with a single 1-cm estradiol implant (OVX+E; n=6) was used.  Detection of nNOS and ERα was performed by dual-label immunocytochemistry.  Total numbers of nNOS neurons and the percentage of nNOS neurons co-expressing ERα were determined in the arcuate nucleus (ARC), preoptic area (POA), ventromedial hypothalamus (VMH), and ventrolateral hypothalamus (VLH).  The mean number of nNOS neurons in the VLH (1524±46) was significantly greater than in the POA (464±15), VMH (417±10), and ARC (210±4).  However, the mean number of nNOS neurons did not differ between the OVX and OVX+E groups in any of the areas studied.  The percentage of nNOS neurons co-expressing ERα was significantly greater in the VLH (41±2) than in the ARC (29±1), POA (22±1), and VMH (12±1).  The percentage of nNOS neurons co-expressing ERα did not differ between the OVX and OVX+E groups in any of the areas studied.  In addition to NO, kisspeptin is also involved in the control of reproduction.  Kisspeptin is critical for puberty onset, stimulates GnRH/LH secretion, and has previously been shown in rodents to contact nNOS neurons.  Thus, the second objective of this study was to examine kisspeptin close-contacts onto nNOS neurons and nNOS close-contacts onto kisspeptin neurons.  Dual-label immunofluorescence for kisspeptin and nNOS was conducted and images were captured at 1 µm intervals along the z-plane in the ARC or POA.  Preliminary observations in a limited number of animals indicate that there are very few close-contacts between kisspeptin and nNOS in either area. In summary, this is the first description of the distribution of nNOS in prepubertal sheep.  nNOS is abundantly expressed in several areas of the ovine hypothalamus, but expression or colocalization with ERα does not seem to change with the presence or absence of estradiol in prepubertal ewes.  Also, we have thus far found little evidence of contacts between nNOS and kisspeptin neurons, suggesting there is little direct interaction between these two neuronal populations.


Nothing to Disclose: MNB, JAL, RBM, RLG, SMH

PP05-4 25933 3.0000 FRI 483 A Characterization of Neuronal Nitric Oxide Synthase Neurons in the Hypothalamus of Prepubertal Sheep 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 11:45:00 AM PP05 7674 11:30:00 AM Neuroendocrinology Poster Preview

Angela Y Liu*, Mohammed Almohaya and David L Kendler
University of British Columbia, Vancouver, BC, Canada


Denosumab has been shown to increase bone mineral density (BMD) in patients with osteoporosis.  Randomized trials report that transitioning from alendronate or zoledronic acid to denosumab (DEN) leads to greater increases in BMD compared to continuing bisphosphonate therapy (1,2).  The effects of switching osteoporosis therapy likely differ between real-world clinic patients and trial patients due to patient characteristics and other factors.

To study the effect of DEN in treatment-naive patients and patients switched from other osteoporosis therapies in a real-world clinic setting, we conducted a retrospective chart review of all patients treated with DEN at an osteoporosis referral centre in Vancouver, Canada.  The study group consisted of all males and females treated with DEN 60 mg SC every six months for at least one year, and in whom baseline and follow-up BMD data were available.  Annual BMD measurements at either hip or spine were performed at the site with the lowest T-score.  Patients were either treatment-naive when starting DEN, or switched from one of four medications: alendronate (ALE), risedronate (RIS), zoledronic acid (ZOL), or teriparatide (TER).  Data analysis was conducted using ANOVA with Bonferroni correction for multiple comparisons.

758 consecutive patients were included: 310 followed at the hip and 448 followed at the spine.  Baseline characteristics were similar between hip and spine follow-up groups.  Treatment-naive and all prior-treatment groups increased BMD on DEN.  Responder analysis showed that 52% and 87% of patients followed at the hip and spine respectively had increased BMD by more than 3% by the end of follow-up at 4 years.  Treatment-naive patients showed a greater increase in hip BMD on DEN compared to patients switching from ZOL after 1 year (p=0.006).  This difference persisted at 3 years (p=0.005).  Treatment-naive patients followed with spine BMD also showed a greater response to DEN compared to those switching from ZOL after 1 year (p=0.007).  Patients switched from ALE, RIS, and TER showed similar increases in hip or spine BMD compared to treatment-naive patients initiating DEN therapy.

Consistent with reported data from clinical trials, DEN increased BMD at hip and spine both in treatment-naive and in patients switching from prior osteoporosis therapy.  The observed increase in BMD was greater than reported in controlled trials (1,3).  Lower adherence to preceding therapy and the use of generic bisphosphonates in clinic patients may account for some of this difference.  There is also greater variability in clinic patient characteristics compared to clinical trial participants.  Our results will inform clinicians regarding anticipated improvements in BMD when transitioning to denosumab.


Disclosure: DLK: Consultant, Amgen, Consultant, Eli Lilly & Company, Consultant, Merck & Co., Consultant, GlaxoSmithKline, Consultant, Astra Zeneca. Nothing to Disclose: AYL, MA

PP01-1 24567 1.0000 FRI 349 A Effects of Denosumab on Bone Mineral Density in Treatment-Naive Patients and Patients Previously Treated with Other Osteoporosis Therapies 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 11:45:00 AM PP01 7687 11:30:00 AM Osteoporosis: What You Had, What You Lost, and What You Gain Poster Preview

Stuart M Sprague*1, Stephen Strugnell2, Joel Z Melnick2, Jay White2, Martin Petkovich3 and Charles W Bishop2
1NorthShore University Health System-University of Chicago, Pritzker School of Medicine, Evanston, IL, 2OPKO Renal, Miami, FL, 3Queens University, Kingston, ON


Vitamin D insufficiency (VDI) is defined as serum total 25-hydroxyvitamin D (25D) below 30 ng/mL in clinical practice guidelines applicable to secondary hyperparathyroidism (SHPT) in chronic kidney disease (CKD; Holick et al 2011; K/DOQI 2003; KDIGO 2009).  Current regimens for correcting VDI in CKD successfully raise serum 25D above 30 ng/mL in some patients but are ineffective in controlling SHPT.  A recent study by Ennis et al (1) suggested that 25D levels significantly greater than 30 ng/mL are required to maximally lower elevated intact parathyroid hormone (iPTH) in CKD. 

   The effects of modified-release calcifediol (MRC) on elevated plasma iPTH and serum markers of bone turnover were examined in two identical, randomized, double-blind, placebo-controlled trials conducted in patients with SHPT (>85 pg/mL), stage 3 or 4 CKD and low serum 25D (10-30 ng/mL).  The trials randomized a total of 429 subjects from 77 US sites, stratified by stage, 2:1 to receive oral MRC or placebo for 26 weeks.  MRC dosing started at 30 μg/d at bedtime and increased to 60 μg/d after 12 weeks if plasma iPTH remained above 70 pg/mL.  Of the 429 subjects, 356 (83%) completed treatment and were grouped according to their 25D levels (0-20, >20-40, >40-60, >60-80 or >80 ng/mL) at the end of treatment (EOT).  Mean EOT plasma iPTH, serum total 1,25-dihydroxyvitamin D (1,25D), and serum calcium (Ca), phosphorus (P) and bone formation/resorption markers, as well as urine Ca and P, were compared among the five groups.    

   Baseline demographics were similar for the five groups with mean age of 63.2-68.7 years, mean serum 25D of 16.5-21.5 ng/mL, mean plasma iPTH of 134.8-156.5 pg/mL and mean eGFR of 30.1-32.3 mL/min/1.73m2.  More than 95% of subjects treated with MRC achieved serum 25D levels of  greater than 30 ng/mL at EOT.  Mean serum 1,25D progressively rose with increasing serum 25D regardless of CKD stage.  Similarly, and independent of CKD stage, mean plasma iPTH, serum collagen type 1 C-telopeptide and serum procollagen type 1 N-terminal propeptide progressively decreased with increasing 25D.   Serum bone-specific alkaline phosphatase decreased with serum 25D increasing up to >40-60 ng/mL.  No increases in mean serum Ca or P, or urine Ca or P were apparent with increasing serum 25D levels. Treatment-emergent adverse events did not associate with serum 25D levels and appeared to be higher at lower 25D levels.

   In conclusion, MRC increased serum 25D and 1,25D, and reduced plasma iPTH and other serum bone markers in patients with stage 3 or 4 CKD, SHPT and VDI with no effect on serum Ca or P, or urine Ca or P.  Decreases in both plasma iPTH and bone markers were maximal at serum 25D levels above 60 ng/mL.  These findings indicate that serum 25D levels recommended for non-CKD patients are insufficient to control SHPT in stage 3 or 4 CKD and that higher, more effective levels (>60 ng/mL) can be safely achieved with daily MRC.


Disclosure: SMS: Advisory Group Member, Opko Pharm, Clinical Researcher, Opko Pharm. SS: Management Position, OPKO Health. JZM: Employee, OPKO. JW: Employee, OPKO Health. MP: Consultant, OPKO Health. CWB: , OPKO Health, Inc..

PP01-3 26734 3.0000 FRI 363 A Treatment Effects of Modified-Release Calcifediol on Bone Markers Suggest Higher 25-Hydroxyvitamin D Levels Are Needed for Adequacy in Patients with Stage 3 or 4 CKD 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 11:45:00 AM PP01 7687 11:30:00 AM Osteoporosis: What You Had, What You Lost, and What You Gain Poster Preview

P D Miller*1, N Pannacciulli2, J P Brown3, E Czerwinski4, B S Nedergaard5, M A Bolognese6, J Malouf7, H G Bone8, JY Reginster9, A Singer10, C Wang2 and R B Wagman2
1Colorado Center for Bone Research, Lakewood, CO, 2Amgen Inc., Thousand Oaks, CA, 3Laval University and CHU de Québec (CHUL) Research Centre, Quebec City, QC, Canada, 4Krakow Medical Center, Krakow, Poland, 5Center for Clinical and Basic Research, Aalborg, Denmark, 6The Bethesda Health Research Center, Bethesda, MD, 7Hospital de la Santa Creu i Sant Pau, Barcelona, Spain, 8Michigan Bone and Mineral Clinic, Detroit, MI, 9University of Liège, Liège, Belgium, 10Georgetown University Medical Center, Washington, DC


Introduction: Denosumab (DMAb) is a RANKL inhibitor that reduces osteoclast number and activity, thus decreasing bone resorption and increasing bone mass. The antiresorptive action of DMAb is associated with transient increases in serum intact parathyroid hormone (iPTH). This might contribute to the effects of DMAb on bone by increasing modeling-based bone formation in a setting of fully inhibited osteoclastic bone resorption [1]. The increase in serum iPTH was greater following DMAb than oral alendronate [2]. Whether this difference also occurs with intravenous (IV) bisphosphonate (BP) has not been reported previously. This analysis compared the effect of DMAb on serum iPTH, as well as serum albumin-adjusted calcium (Ca), with an IV BP, zoledronic acid (ZOL), in subjects enrolled in a substudy of a randomized double-blind trial in postmenopausal women with osteoporosis previously treated with oral BP.

Methods: This was a 12-month, multicenter, randomized, double-blind, double-dummy study in 643 postmenopausal women aged ≥ 55 years who had received oral BP for ≥ 2 years and had a bone mineral density (BMD) T-score ≤ –2.5 at the lumbar spine, total hip, or femoral neck. Subjects were randomized 1:1 to DMAb 60 mg subcutaneously (SC) every 6 months (Q6M) + placebo (IV once) or ZOL 5 mg IV once + placebo (SC Q6M) for 12 months, and received daily calcium (≥ 1000 mg) and vitamin D (≥ 800 IU). A subset of 117 subjects (61 DMAb, 56 ZOL) was enrolled to have serum iPTH and serum albumin-adjusted Ca evaluated at baseline, day 10 (Ca only), and months 1, 3, 6, 6+10 days (Ca only), 7, 9, and 12.

Results: Significant median percentage increases from baseline in serum iPTH were observed at months 1 (33.9%), 3 (19.7%), and 9 (10.3%) with DMAb but only at month 1 with ZOL (15.1%; all p<0.05). Median percentage changes from baseline in serum iPTH were significantly greater at months 3 and 9 with DMAb (19.7% and 10.3%, respectively) vs ZOL (–5.6% and –8.9%, respectively) (p<0.05). There were no significant percentage changes from baseline in serum albumin-adjusted Ca in either group (except at month 12 in the ZOL group [+1.0%, p<0.01]), and no difference was observed between treatment groups at all visits.

Conclusion: In postmenopausal women with osteoporosis previously treated with oral BP, DMAb treatment was associated with greater transient increases in serum iPTH vs ZOL and greater gains in BMD at all measured skeletal sites. These data help support the hypothesis that the unique mechanism of action of DMAb is due to its direct effect to fully and rapidly inhibit osteoclastic activity at cortical and trabecular bone and its indirect effect to increase serum iPTH.


Disclosure: PDM: Advisory Group Member, Lilly USA, LLC, Advisory Group Member, AgNovos, Advisory Group Member, Amgen, Advisory Group Member, Alexion, Speaker Bureau Member, Radius Health, Speaker Bureau Member, Amgen, Speaker Bureau Member, Alexion, Research Funding, Takeda, Research Funding, Roche Diagnostics, Research Funding, Radius Pharma, Research Funding, Novo Nordisk, Research Funding, Novartis Pharmaceuticals, Research Funding, NBHA, Research Funding, Merck Serrano, Research Funding, Merck & Co., Research Funding, Lilly USA, LLC, Research Funding, Immunodiagnostics, Research Funding, Boehringer Ingelheim, Research Funding, Amgen, Research Funding, Alexion, Advisory Group Member, Merck & Co., Advisory Group Member, Radius Pharma, Advisory Group Member, Roche. NP: Employee, Amgen, Employee, Amgen, Employee, Amgen. JPB: Advisory Group Member, Amgen, Study Investigator, Amgen, Speaker, Amgen, Advisory Group Member, Eli Lilly & Company, Study Investigator, Eli Lilly & Company, Speaker, Eli Lilly & Company, Advisory Group Member, Merck & Co.. EC: Principal Investigator, Amgen, Lecturer, Amgen. BSN: Principal Investigator, Amgen. MAB: Study Investigator, Amgen, Speaker Bureau Member, Amgen, Study Investigator, Pfizer, Inc., Study Investigator, Sanofi, Study Investigator, Lilly USA, LLC. JM: Principal Investigator, Lilly España, Principal Investigator, Amgen, Speaker, Gruenenthal. HGB: Speaker, Amgen, Consultant, Amgen, Investigator, Amgen, Consultant, Merck & Co., Investigator, Merck & Co., Investigator, NPS/Shire. JR: Research Funding, Teva, Research Funding, Rottapharm, Research Funding, Merck Sharp & Dohme, Research Funding, Bristol-Myers Squibb, Speaker, Nolver, Speaker, Novo Nordisk, Speaker, Nycomed, Speaker, Theramex, Speaker, Analis, Speaker, Zodiac, Speaker, Ebewee Pharma, Speaker, Teva, Speaker, Teijin, Speaker, GlaxoSmithKline, Speaker, Roche, Speaker, Servier, Speaker, Novartis Pharmaceuticals, Speaker, Genevrier, Speaker, IBSA, Speaker, Rottapharm, Speaker, Lilly USA, LLC, Speaker, Merck Sharp and Dohme, Consultant, UCB, Consultant, Theramex, Consultant, NPS, Consultant, Nycomed, Consultant, Merckle, Consultant, Roche, Consultant, GlaxoSmithKline, Consultant, Amgen, Consultant, Lilly USA, LLC, Consultant, Negma, Consultant, Novartis Pharmaceuticals, Consultant, Servier, Research Funding, Lilly USA, LLC, Research Funding, Novartis Pharmaceuticals, Research Funding, Roche, Research Funding, GlaxoSmithKline, Research Funding, Amgen, Research Funding, Servier, Consultant, Wyeth. AS: Principal Investigator, Amgen, Advisory Group Member, Amgen, Speaker Bureau Member, Amgen, Advisory Group Member, Eli Lilly & Company, Educational video, Actavis, Consultant, Mission Pharmacal. CW: Employee, Amgen, Employee, Amgen, Employee, Amgen. RBW: Employee, Amgen, Employee, Amgen, Employee, Amgen.

PP01-4 23915 4.0000 FRI 351 A Denosumab Treatment Is Associated with Greater Transient Increases in Serum Intact Parathyroid Hormone Concentrations Compared with Zoledronic Acid: Results from a Study in Postmenopausal Women with Osteoporosis Previously Treated with Oral Bisphosphonates 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 11:45:00 AM PP01 7687 11:30:00 AM Osteoporosis: What You Had, What You Lost, and What You Gain Poster Preview

Wasana Pratchayasakul*, Sivaporn Sivasinprasasn, Piangkwan Sa-nguanmoo, Cicely Proctor, Sasiwan Kerdphoo, Nipon Chattipakorn and Siriporn C Chattipakorn
Chiang Mai University, Chiang Mai, Thailand


Obesity and estrogen deprivation are associated with increased oxidative stress level (1,2).  In the brain, we previously showed that both obesity and estrogen deprivation caused a reduction of dendritic spine density and was associated with cognitive decline (3).   Moreover, either estrogen or a dipeptidyl peptidase 4 (DPP-4) inhibitor therapy significantly decreased brain oxidative stress and increased dendritic spine density in obese and estrogen-deprived rats (4,5).  Growing evidence shows that both obesity and estrogen deprivation increase risks of cardiac ischemia (6).  However, the effects of cardiac ischemia-reperfusion injury (cardiac IR) on brain oxidative stress and dendritic spine density in rats under obese and estrogen-deprived conditions have never been investigated.   We hypothesized that cardiac IR aggravates brain oxidative stress and decreases dendritic spine density in ovariectomized obese rats, and that estrogen and DPP-4 inhibitor treatments attenuates these adverse effects.  Female rats were divided into sham (S) and bilateral ovariectomized (O) groups.  Sham rats were fed with normal diet for 12 weeks (NDS) and ovariectomized rats were divided into two subgroups to be fed with either a normal diet (NDO) or high fat diet (HFO) for 12 weeks.  At week 13, ovariectomized rats in each dietary group (NDO and HFO) were treated with either a vehicle (V), 50 µg/kg estradiol (E) or 3 mg/kg DPP-4 inhibitor (Vildagliptin; Vil) for 4 weeks (n=5/subgroup).  Finally, all rats were subject to cardiac IR by left anterior descending artery occlusion for 30 min, followed by 120-min reperfusion.  Brains were rapidly removed for determining the levels of brain oxidative stress and dendritic spine density.  We found that ovariectomy increased brain oxidative stress and decreased dendritic spine density, and obesity aggravated that impairment in ovariectomized rats.  Interestingly, we found that cardiac IR aggravated brain oxidative stress and decreased dendritic spine density in NDS and NDO rats (p<0.05), but not in HFO rats (7.71+0.33, 6.53+0.21, 3.58+0.38, 4.42+0.37, 5.11+0.39 and 4.27+0.19 spines/20µm for NDS, NDO, HFO, NDSIR, NDOIR and HFOIR groups, respectively).  Estrogen and DPP-4 inhibitor treatments significantly reduced brain oxidative stress and increased dendritic spine density in both NDO rats and HFO rats with cardiac IR (5.11+0.39,7.13+0.11,7.07+0.21, 4.27+0.19, 6.85+0.14 and 6.82+0.21 spines/20µm for NDO, NDOVil, NDOE, HFO, HFOVil and HFOE groups, respectively, p<0.05).  Our findings suggest that cardiac IR aggravates brain oxidative stress, leading to decreased dendritic spine density in ND rats.  However, cardiac IR injury did not aggravate the severity in HF rats, which could be due to the severity already at its maximum in these rats before cardiac IR.  Nevertheless, estrogen and DPP-4 inhibitor treatments effectively attenuate these impairments in all groups.


Nothing to Disclose: WP, SS, PS, CP, SK, NC, SCC

PP03-1 24591 1.0000 FRI 656 A Estrogen and DPP-4 Inhibitor Reduced Brain Oxidative Stress and Increased Dendritic Spine Density in Ovariectomized Obese and Non-Obese Rats with Cardiac Ischemia-Reperfusion Injury 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 11:45:00 AM PP03 7694 11:30:00 AM Cardiometabolic Risk - Salt, Fat and Sex Poster Preview

Pongpan Tanajak, Hiranya Pintana, Natthaphat Siri-Angkul, Siriporn C Chattipakorn, Nipon Chattipakorn and Nattayaporn Apaijai*
Chiang Mai University, Chiang Mai, Thailand


Long-term high-fat diet (HFD) consumption leads to obese-insulin resistance and fibroblast growth factor 21 (FGF21) resistance, resulting in left ventricular (LV) dysfunction.  Evidence demonstrates that anti-diabetic drug dipeptidyl peptidase-4 (DPP-4) inhibitor such as Vildagliptin as well as lifestyle modification such as caloric restriction (CR) could improve metabolic regulation and LV function in obese-insulin resistant subjects.  However, the comparative effects of CR vs. Vildagliptin, and the role of combined therapy on metabolic disturbance, cardiac autonomic balance, and LV function in obese-insulin resistant condition have not been investigated.  We tested the hypothesis that combined CR and Vildagliptin therapy exerts better efficacy in attenuating metabolic disturbance, cardiac autonomic imbalance, and LV function in obese-insulin resistant condition than a monotherapy.  Thirty male Wistar rats were divided into 2 groups to received normal diet (ND) or HFD for 12 weeks.  Then, rats in the HFD group were divided into 4 subgroups.  Each subgroup received one of the following treatment: vehicle (HFV), CR diet (60% of energy of food intake during the first 12 weeks; HFR), Vildagliptin (3 mg/kg/day; HFVil), or CR and Vildagliptin (HFRVil) for 4 weeks.  The ND rats were further continued with ND consumption and received a vehicle (NDV) for 4 weeks.  Metabolic profiles, blood pressure (BP), oxidative stress, cardiac mitochondrial function, heart rate variability (HRV), LV function, and FGF21 sensitivity were determined.  The results showed that rats fed with HFD developed obesity, insulin resistance, dyslipidemia, increased plasma FGF21 levels (2.91 ± 0.54 vs. 0.80 ± 0.08 pg/ml; P=0.003; n=6 per group), decreased HRV, impaired LV function, cardiac mitochondrial dysfunction, and impaired cardiac tissue FGF21 sensitivity by decreased cardiac mitochondrial fatty acid β-oxidation and anti-apoptosis signaling pathways.  Rats in the HFR, HFVil and HFRVil groups had restored metabolic function by decreased dyslipidemia, reduced serum and cardiac MDA (P<0.001; n=6 per group), plasma FGF21 levels (P<0.001; n=6 per group), cardiac mitochondrial ROS production (P<0.001; n=6 per group), and also increased cardiac mitochondrial FAO signaling pathways when compared with the HFV group.  Although only rats received CR diet had reduced body weight, rats received Vildagliptin showed better improvement than the CR rats in decreasing BP and heart rate, improving HRV and LV function, reducing cardiac mitochondrial dysfunction (P<0.001; n=6 per group), and increasing anti-apoptosis signaling pathways.  These findings indicate that DPP-4 inhibitor exerts better cardioprotection than CR.  This is due to the effective preservation of cardiac FGF21 sensitivity and cardiac mitochondrial function by DPP-4 inhibitor in obese-insulin resistant rats.


Nothing to Disclose: PT, HP, NS, SCC, NC, NA

PP03-2 24392 2.0000 FRI 654 A DPP-4 Inhibitor Exerts Better Cardioprotection Than Caloric Restriction By Attenuating Cardiac Mitochondrial Dysfunction and Improving FGF21 Sensitivity in Obese-Insulin Resistant Rats 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 11:45:00 AM PP03 7694 11:30:00 AM Cardiometabolic Risk - Salt, Fat and Sex Poster Preview

Cleyton C. Domingues1, Nabanita Kundu2, Neeki Ahmadi2 and Sabyasachi Sen*3
1The George Washington University, Washington, DC, 2George Washington University, 3The George Washington University, DC


MSCs are undifferentiated, multipotent cells. We have previously showed that High Glucose (HG, 25mM) exposure promotes adipogenic differentiation, increased formation of ROS and decreased cellular oxygen consumption rate (OCR,using Seahorse). HG exposure also upregulated mRNA expression of adipogenic (PPARG, FABP-4, CEBP alpha and beta) and inflammatory (IL-6 and TNF alpha) genes. In this study, in order to reduce intracellular superoxide presence, we used GFP-containing Adenovirus constructs to upregulate both mitochondrial and cytosolic antioxidants (SOD2 and SOD1, respectively) and used GFP gene as a control. We showed both SOD1 and SOD2 upregulation reduced intracellular superoxide presence and improved OCR in presence of HG environment. In addition, the upregulation of IL-6 and TNFa was prevented. Next, we delivered the eGFP, SOD1 and SOD2 upregulated MSCs intra-peritoneally to DIO (60% high-fat diet) C57BL/6J mice. Previously to MSC delivery, all mice presented fasting blood glucose levels of 200mg/dl or above (measured from the tail vein using a glucometer). We confirmed homing-in of eGFP labeled MSC to different inflamed fat pockets, particularly pericardial and omental fat by direct imaging. Our results indicate that mice receiving SOD2 MSCs improve glucose tolerance (GTT) at 4 weeks (increased area under the curve) compared to SOD1 and GFP with progressive reduction in fat mass. We are currently processing fat depots and serum samples from MSC delivered mice, for qPCR, Western Blot and ELISA to demonstrate reduction of local and systemic inflammation. In summary, HG evokes superoxide generation, OCR reduction and adipogenic differentiation. Upregulation of superoxide dismutase, particularly mitochondrial, quenches excess of intracellular superoxide and improves MSCs respiration. Delivery of superoxide dismutase using MSCs as a gene delivery vehicle in DIO mice improved glucose tolerance. We conclude that delivery of dismutases using MSCs to the inflamed adipocyte depots may be the key to suppression of adipocyte mediated inflammation and may be a novel yet safe therapeutic tool to combat obesity associated diabetes and impaired glucose tolerance.


Nothing to Disclose: CCD, NK, NA, SS

PP03-3 24982 3.0000 FRI 658 A Modified Human Mesenchymal Stromal Cells (MSCs) Improve Glucose Tolerance Test in Diet Induced Obese (DIO) Diabetic Mouse Model 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 11:45:00 AM PP03 7694 11:30:00 AM Cardiometabolic Risk - Salt, Fat and Sex Poster Preview

Indrani Sinha-Hikim*1, Theodore C Friedman2, Mark Falz1, Victor Chalfan3, Isai Rea3, Desean L. Lee1, Carl Sims3 and Amiya P Sinha-Hikim1
1Charles R. Drew University, Los Angeles, CA, 2Charles R. Drew Univeristy of Medicine and Science, Los Angeles, CA, 3Charles R. Drew University, Endocrinology


Background and Objective: Cigarette smoking is an important risk factor for diabetes, cardiovascular disease, and nonalcoholic fatty liver disease. The health risk associated with smoking can be exaggerated by obesity. Smoking may also trigger cardiomyocyte (CM) apoptosis. Given that CM apoptosis has been implicated as a potential mechanism in the development of cardiomyopathy and heart failure, we characterized the key molecular component of the effector pathways of nicotine plus high-fat diet (HFD)-induced CM apoptosis.

Experimental Design: Adult C57BL6 male mice were fed a normal diet (ND) or HFD and received twice-daily IP injection of nicotine (0.75 mg/kg BW) or saline for 16 weeks. An additional group nicotine-treated mice on a HFD received twice daily IP injections of 1 mg/kg BW of mecamylamine, a nonselective nicotinic acetylcholine receptor antagonist, for 16 weeks.

Results:HFD alone led to ventricular lipid accumulation compared with mice fed with ND with or without nicotine. Nicotine treatment effectively prevented such HFD-induced ventricular lipid accumulation. A very low incidence of CM apoptosis, expressed as the percentage of TUNEL-positive nuclei per total (apoptotic plus non-apoptotic) nuclei was noted in mice fed ND with or without nicotine or HFD alone. However, combined with a HFD, nicotine led to a significant (P<0.05) increase in CM apoptosis. Mecamylamine treatment fully prevented nicotine and HFD-induced CM apoptosis. Induction of CM apoptosis was associated with increased oxidative stress and activation of caspase 2-mediated intrinsic pathway signaling coupled with inactivation of AMP-activated protein kinase (AMPK). Furthermore, nicotine treatment significantly (P<0.05) blunted the inhibitory effects of HFD on fibroblast growth factor 21 and silent information regulator 1.

Conclusion: We conclude that nicotine when combined with a HFD triggers CM apoptosis though generation of oxidative stress and inactivation of AMPK together with activation of the caspase 2-mediated intrinsic apoptotic signaling. The clinical implication of this study is that smiking plus a HFD may lead to heart failure and other forms of cardiomyopathy.


Nothing to Disclose: IS, TCF, MF, VC, IR, DLL, CS, APS

PP03-4 24959 4.0000 FRI 655 A Nicotine Plus a High-Fat Diet Triggers Cardiomyocyte Apoptosis in Male Mice By Inhibiting AMP-Activated Protein Kinase Independent of Fibroblast Growth Factor 21 and Silent Information Regulator 1 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 11:45:00 AM PP03 7694 11:30:00 AM Cardiometabolic Risk - Salt, Fat and Sex Poster Preview

Mohammed Al-Sofiani*1, Shabnam Rehman2 and Howard A Lippes3
1University at Buffalo- CHS, Williamsville, NY, 2University at Buffalo-CHS, Buffalo, 3University at Buffalo-CHS, Williamsville, NY


Introduction: Diabetics with severe insulin resistance present a challenge in achieving glycemic control. The large amount of insulin and number of injections required in these cases are burdensome and can lead to poor compliance. Here, we report our experience with the use of five-fold concentrated regular insulin (U-500R) via continuous subcutaneous insulin infusion (CSII).

 Methods: We retrospectively reviewed medical records of eight patients who have used U-500R via CSII. Two patients were excluded from the statistical analysis because of the unavailability of a follow-up hemoglobin A1c (HbA1c) in one patient and discontinuation of CSII in the second patient due to contact dermatitis.

Results: All patients had type II diabetes with an average weight of 113.25 Kg and daily insulin dose of 2.75 U/Kg at baseline. After initiating the U-500R via CSII, the mean HbA1c decreased from 9.07% to 8.4%. The reduction in HbA1c was associated with a significant reduction in the average total daily dose of insulin from 303.33 U/day to 234 U/day (P= 0.02).

Conclusion: The use of U-500R via CSII appears to be an effective treatment option in achieving better glycemic control with less insulin in type II diabetics with severe insulin resistance. The ability to infuse precise amounts of U-500R at a continuous basal rate with boluses makes the U-500R via CSII a suitable option for type II diabetics who have not achieved their glycemic goals. Prospective studies to assess long-term effectiveness, side effects and potential cost savings with the use of U-500R via CSII are necessary.


Nothing to Disclose: MA, SR, HAL

PP12-1 27618 1.0000 FRI 684 A Effectiveness of U-500 Regular Insulin Via Continuous Subcutaneous Insulin Infusion Using V-Go 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 11:45:00 AM PP12 7710 11:30:00 AM Fresh Thoughts on Diabetes Treatment Poster Preview

Farid Saad1, Ahmad Haider*2, Karim Sultan Haider2, Gheorghe Doros3 and Abdulmaged M Traish4
1Bayer Pharma AG, Berlin, Germany, 2Private Urology Practice, Bremerhaven, Germany, 3Boston University School of Public Health, Boston, MA, 4Boston University School of Medicine, Boston, MA



A registry was established to assess long-term effectiveness and safety of TU in a urological setting in comparison to an untreated hypogonadal control group.

Material and Methods:

Observational, prospective, cumulative registry study in 656 men (age: 60.72 ± 7.15 years) with total testosterone (T) levels below 12.1 nmol/L and symptoms of hypogonadism. 360 men received parenteral TU 1000 mg/12 weeks following an initial 6-week interval for up to 10 years. 296 men had opted against TTh and served as controls. Median follow-up in both groups was 7 years. Measurements were taken at least twice a year, and 8-year data were analysed. Mean changes over time between the two groups were compared by means of a mixed effects model for repeated measures with a random effect for intercept and fixed effects for time, group and their interaction. Changes were adjusted for age, weight, waist circumference, blood pressure, and lipids to account for baseline differences between the two groups.


In the T group, 113 men (31.4%) had type 2 diabetes mellitus (T2DM), in the control group, 114 (38.5%).

Across all patients, fasting glucose (mmol/L) decreased from 5.65 ± 0.7 to 5.23 ± 0.05 (p<0.0001) in the T group and remained stable from 5.57 ± 0.36 to 5.56 ± 0.34 (NS) in the controls. The model-adjusted estimated difference between groups at 8 years was -0.61 mmol/L (p<0.0001).

HbA1c (%) decreased from 6.87 ± 1.42 to 5.59 ± 0.44 in the T group and increased from 6.09 ± 1.22 to 6.38 ± 1.44 in the controls. The difference between groups at 8 years was -1.83% (p<0.0001 for all).

The triglyceride:HDL ratio, a surrogate marker for insulin resistance, decreased from 5.5 ± 1.99 to 2.65 ± 0.67 (p<0.0001) in the T group and from 6.48 ± 3.61 to 5.76 ± 3.56 (p=0.0102) in the controls. The difference between groups at 8 years was -1.24 mmol/L (p<0.0001 for all).

The TyG index, another surrogate marker for insulin resistance, decreased from 4.13 ± 0.1 to 3.95 ± 0.02 in the T group and increased from 4.1 ± 0.09 to 4.13 ± 0.09 in the controls. The difference between groups at 8 years was -0.24 (p<0.0001 for all).

Medication adherence in the testosterone group was 100 per cent as injections were administered in the office.

There were two deaths in the T group and 21 deaths in the control group. No patient dropped out.


Long-term TTh with TU in an unselected cohort of hypogonadal men resulted in improvements in glycaemic control, whereas there was a worsening except for fasting glucose in untreated controls. Long-term TU was well tolerated and excellent adherence suggested a high level of patient satisfaction.


Disclosure: FS: Employee, Bayer Schering Pharma. AH: Investigator, Bayer Schering Pharma. GD: Coinvestigator, Bayer Schering Pharma. Nothing to Disclose: KSH, AMT

PP12-2 26390 2.0000 FRI 686 A Effects of Long-Term Therapy with Testosterone Undecanoate Injections (TU) on Glycaemic Control in Hypogonadal Men: Real-Life Data from a Registry Study 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 11:45:00 AM PP12 7710 11:30:00 AM Fresh Thoughts on Diabetes Treatment Poster Preview

Andrew Orville Paulus*1, Jeffrey Adam Colburn2, Jack Edward Lewi2, Irene Folaron2, Sky Denniston Graybill2, Richard Price Davis2, Darrick James Beckman2 and Mark Windell True2
1Wright State University, Dayton, OH, 2San Antonio Military Medical Center, San Antonio, TX



The number of patients using U-500 regular insulin has significantly increased in recent years.  These patients are severely insulin resistant requiring high doses of insulin to achieve glycemic control.  However, it has been observed that a patient’s insulin requirements may dramatically decrease upon admission to the hospital. The medical literature is sparse on this issue, with no formal published guidelines. Therefore, we sought to systematically investigate this phenomenon in our institution.


We performed a retrospective chart review of patients with U-500 insulin outpatient regimens who were admitted to the San Antonio Military Medical Center over a five-year period. Each patient’s outpatient total daily dose (TDD) of insulin was compared to the average inpatient TDD. The outpatient estimated average glucose (eAG) was calculated from the HgbA1c and compared to the average inpatient glucose level.


We collected data on 27 patients with a total of 62 separate admissions. The average age was 64.4 years with a mean body mass index of 38.9 kg/m2 and eAG of 203mg/dl [74 – 109] (HgbA1c of 8.7% [0 – 5.6]). All patients were converted from U-500 to various U-100 insulin regimens upon admission. The average TDD of insulin received in the hospital was lower than their outpatient TDD, 91 units vs. 337 units (p < 0.001) or 27% of their outpatient insulin dose. Overall, 89% of patients received ≤ 50% of their outpatient TDD while in the hospital. The average inpatient glucose was slightly higher than the outpatient eAG, 234 mg/dl vs. 203 mg/dl (p < 0.003).


Patients using U-500 insulin routinely have their TDD of insulin reduced upon admission in our institution. Possible factors for decreased insulin requirements include controlled hospital diet, NPO status, and strict adherence to insulin injections by nursing staff. U-500 insulin dosing is prone to errors in the hospital setting, so conversion to U-100 insulin with a dose reduction is a preferred option. Despite a significant reduction in insulin TDD, these patients had clinically similar glucose levels. Therefore, we propose that it is reasonable to convert U-500 patients to U-100 insulin on admission with starting doses between 25 to 50% of their home TDD. Patients’ glucose should be closely monitored and insulin titrated as needed to maintain adequate glucose control.


Patients taking U-500 insulin as an outpatient should be converted to a U-100 basal-bolus regimen at 25 to 50% of their home TDD upon hospital admission. Further prospective trial data is needed to best evaluate the ideal approach to this situation.


Nothing to Disclose: AOP, JAC, JEL, IF, SDG, RPD, DJB, MWT

PP12-3 25780 3.0000 FRI 683 A Evaluation of Total Daily Dose and Glycemic Control for Patients on U-500 Insulin Admitted to the Hospital 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 11:45:00 AM PP12 7710 11:30:00 AM Fresh Thoughts on Diabetes Treatment Poster Preview

Maria Burnett*1, H. Omer Ikizler1, Annis Morison Marney2 and Matthew P. Gilbert3
1The University of Vermont College of Medicine, 2Frist Clinic, 3The University of Vermont College of Medicine, South Burlington, VT


Introduction:  The development of continuous subcutaneous insulin infusion (CSII) and continuous glucose monitors (CGM) promises to improve quality of life and outcomes in patients with Type 1 Diabetes Mellitus (T1DM). CSII and CGM can improve HbA1c and reduce hypoglycemia versus multiple daily injections (MDI) (1-4). However, there are few data regarding quality of life. There are also no strict criteria regarding when to use CSII ± CGM despite their cost. To elucidate who benefits most from CSII ± CGM, we investigated demographic and medical characteristics of patients with T1DM who use CSII ± CGM versus MDI. We hypothesized that CSII ± CGM users would be younger, geographically closer to the clinic, have lower HbA1c, and have fewer complications than MDI users. 

Methods:  In a retrospective cohort study, we reviewed the records of patients with T1DM treated at the University of Vermont Medical Center Endocrinology Clinic. We grouped patients as those using CSII, those using CSII + CGM, and those using MDI. We examined age, sex, duration of diabetes, distance from clinic, HbA1c within 12 months, and presence of diabetic complications.  Results:  We reviewed 314 patient records (39% of the clinic’s T1DM population ). 74% of patients reviewed (n=232) used CSII ± CGM.  Groups did not differ in age or sex. Patients using CSII had increased duration of DM compared to patients using MDI or CSII + CGM (F(2,309)=6.967, p=0.0011). Patients who used CSII ± CGM tended to live further away (ß=0.01 per mile from clinic, SE=0.01, p=0.04).  HbA1c was significantly higher in the MDI group (8.53% ± 2.13) versus CSII with and without CGM (adjusted mean=7.34%, t=-4.602, p=0.001; adjusted mean=7.79%, t=3.327, p=0.003, respectively). Duration of disease was significantly associated with cardiovascular disease, retinopathy, peripheral neuropathy, and renal disease.  CSII ± CGM was not associated with decreased incidence of diabetic complications.  

Conclusions:  Contrary to our expectations, patients with CSII ± CGM lived further from clinic and did not have lower occurrences of comorbidities and complications, despite having lower HbA1c levels.  Patients living far away may either be selected by their provider or request CGM ± CSII for better glycemic control to decrease frequency of visits to the endocrinologist. Patients may begin CGM ± CSII too late in the disease course to avert complications, thus having multiple comorbidities and complications despite improved glycemic control as measured by HbA1c.


Nothing to Disclose: MB, HOI, AMM, MPG

PP12-4 27502 4.0000 FRI 688 A Continuous Insulin Delivery Via Insulin Pump Versus Multiple Daily Injections in Type 1 Diabetes  Mellitus: Patient Demographics and Clinical Outcomes at the University of Vermont Medical Center Endocrinology Clinic 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 11:45:00 AM PP12 7710 11:30:00 AM Fresh Thoughts on Diabetes Treatment Poster Preview

Michael Scott Irwig*1, Adrienne Hancock2 and Kayla Childs2
1George Washington Univ, Washington, DC, 2George Washington University


Background:Testosterone therapy for transgender men (female-to-males) is associated with a deepening of the voice to a range perceived as male. There is sparse prospective data regarding the timing and degree of voice changes associated with testosterone therapy in this population.

Methods:Seven transgender men (18-39 years old) naïve to cross-sex hormone therapy and voice therapy were recruited from an outpatient academic endocrinology practice. Each subject underwent prospective voice testing at baseline and following 3, 6 and 9 months after therapy with intramuscular testosterone enanthate or cypionate injections (50-100 mg every 2 weeks).

Results:All subjects showed a dramatic increase in serum testosterone levels at 3, 6 and 9 months consistent with medication use. Baseline fundamental frequencies during a reading task ranged from 134-183 Hz; four voices were consistent with female norms and three were in the “gender neutral” or high end of typical male range. The decline in fundamental frequency over 9 months ranged from 15-74 Hz with an average of 44 Hz. The average decline was 19 Hz between 0-3 months, 19 Hz between 3-6 months and 8 Hz between 6-9 months. Six out of seven subjects had a significant decline within the first 3 months and four out of seven subjects had the largest decline during this period. At 9 months six out of seven subjects had a male range voice and one out of seven had a “gender neutral” voice.

Conclusions:Transgender men show a heterogeneity of baseline fundamental frequencies as well as a heterogeneity of responses to testosterone therapy. Most transgender men can expect significant and meaningful changes in voice within six months of starting testosterone therapy, but the rate and degree of change over the first nine months will vary by individual.


Nothing to Disclose: MSI, AH, KC

PP10-1 24016 1.0000 FRI 134 A Testosterone Therapy and the Female-to-Male Transgender Voice: A Prospective Study 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Reproductive Endocrinology Friday, April 1st 11:45:00 AM PP10 7725 11:30:00 AM Recent Developments in Disorders of Sex Development (DSD) and Transgender Medicine Poster Preview

Gloria Beatriz Aranda Velazquez*1, Mireia Mora Porta2, Felicia Alexandra Hanzu2, Josep Vera3, Emilio Ortega3 and Irene Halperin2
1Hospital Clinic, Barcelona, Barcelona, Spain, 2HCB/IDIBAPS/CIBERDEM, Barcelona, Spain, 3Hospital Clinic, Barcelona


Introduction: In transsexualism, the extreme situation of gender identity disorder (GID) defined as an intense and persistent discomfort with the assigned sex, cross-sex hormone treatment (CHT) both improves and impairs several surrogate cardiovascular risk markers in male to female (MtoF) and female to male (FtoM). Few randomized trials on CHT with long follow-up and control group are available, and present evidence is inconsistent. We here assess changes in metabolic and cardiovascular risk after 12 months CHT compared with biological sex.

Methods: Prospective observational study, including 29 GID individuals (20 FtoM/9 MtoF), attended in the Gender Identity Disorder Unit (UTIG) of Hospital Clinic from July 2012 to November 2013, who accepted to participate. CHT in FtoM consisted of intramuscular testosterone undecanoate (1000 mg every 2-3 months) except for one who received transdermal testosterone (50mg/day). All MtoF received estradiol valerate 2-4mg/day; cyproterone acetate (25-50mg/day) was associated in some of them. Contraindications for CHT were ruled out; subjects had not previously received hormonal treatment, and had no history of cardiovascular disease or HIV. Anthropometric, hormonal, metabolic and coagulation parameters were assessed at baseline, and at 6 and 12 months of CHT. A substudy in FtoM included body composition by DEXA, endothelial dysfunction by flow-mediated dilation (FMD) and Intima-media thickness (IMT) by carotid ultrasound at baseline, 6 and 12 months.

Results: At baseline, FtoM had higher percentage of total fat mass, LH, estradiol and SHBG than MtoF, while MtoF presented higher HOMA, hemoglobin, hematocrit and testosterone than FtoM. Changes after 6 and 12 moths of CHT: FtoM -Anthropometry: increased BMI (p: 0.001). Hormonal: decreased LH (p: 0.008), SHBG (p<0.001) and increased testosterone (p<0.001). Metabolism: increased total cholesterol (p: 0.043), LDL (p: 0.019) and triglycerides (p: 0.001), and decreased HDL (p: 0.035); increased homocysteine (p: 0.003) and leucocytes (p:<0.001). Coagulation: increased hemoglobin (p<0.001) and hematocrit (p<0.001) and decreased platelets (p: 0.011). MtoFAnthropometry: increased BMI (p: 0.015) and decreased waist to hip ratio (p: 0.050). Hormonal: decreased testosterone (p: 0.003) and increased FSH (p: 0.035), SHBG (p: 0.002) and prolactin (p: 0.016). Metabolism: decreased triglycerides (p: 0.019) and increased HDL (p: 0.019). Coagulation: decreased hemoglobin (p: 0.023), hematocrit (p: 0.019), platelets (p: 0.021) and ICAMs (p: 0,014). Substudy of body composition in FtoM: decreased total fat mass (p: 0.050), increased total lean mass (p: 0.007) and decreased ginecoid fat distribution (p: 0.008);  no changes in FMD or IMT.

Conclusion: CHT in FtoM has negative effects that could increase the cardiovascular risk, whereas beneficial changes were observed in MtoF.


Nothing to Disclose: GBA, MM, FAH, JV, EO, IH

PP10-2 25773 2.0000 FRI 135 A Effects of Sex Steroids on Cardiovascular Risk Profile in Individuals with Gender Identity Disorder with Cross-Sex Hormone Treatment 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Reproductive Endocrinology Friday, April 1st 11:45:00 AM PP10 7725 11:30:00 AM Recent Developments in Disorders of Sex Development (DSD) and Transgender Medicine Poster Preview

Maria Bernarda Estevez*1, Patricia Teofilo Monteagudo2, Kelly Christina Oliveira1 and Ieda T N Verreschi3
1UNIFESP, Sao Paulo, Brazil, 2Ministério de Educacao e Cultura, Brazil, Sao Paulo, SP, Brazil, 3UNIFESP, Sao Paulo SP, Brazil


INTRODUCTION: Turner Syndrome (TS) was clinically described in 1938 by Henry Turner.  In 1954 Décourt et cols related the clinical characteristics to the absence of Barr corpusculae. Its cause is the presence of a single functioning X chromosome. Its karyotypes include monosomy, mosaicisms, and structural disruptions of the second sex chromosome.  The possibilities of clinic characteristics are wide. The Quality of life (QoL) associates welfare and capability to live to the fullest. OBJECTIVES: To compare the clínical/laboratory control and QoL on groups with/without TS. HYPOTHESIS: TS patients in treatment may have different scores than normal controls in QoL. METHODS: From Dec-2013 to Dec-2014, 90 participants were recruited from an Endocrinology outpatient clinic of the Federal University in Sao Paulo, Brazil: 48 with TS (TW) and 42 without (CW).  They were of age (18 and older). Participants filled the SF36 questionnaire and blood was drawn in the follicular stage measure oestradiol (E2, normal range 12,5-166 pg/ml in follicular stage), progesterone (P4, normal range 0,2-1,5 ng/dl in follicular stage), LH (normal, 2,4-12,6 mUI/ml in follicular stage, 7,7-58,5 UI/ml in menopause), FSH (3,5-12,5 follicular stage 25,8-134,8 in menopause), SHBG (normal 32,4-128 nmol/l for ages 20-49), SDHEA (33,7-339 ug/dl for ages 10-44) by ECLIA.  Testosterone (10-38 ng/dl Tanner 5) was dosed by LC MS/MS RESULTS:   Age and schooling was similar between groups. The most common occupations were health worker, administration or education in TW, and health worker or cashier in CW.  Most participants were Catholic or Evangelic.  Of the cases 39/48   took Hormonal replacement therapy, mostly transdermal (23/39). TW and CW scored similarly on SF 36 questionnaire. CW had higher oestradiol (105,6 ± 91,5 pg/ml in CW and 50,1± 67,2 mg/ml in TW, p< 0,01) and lower FSH than TW (11 ± 21,7 mUI/ml in CW and 65,6 ± 46,5 mUI/ml in TW, p<0,01). Concentrations of P4, LH, SHBG or SDHEA were alike  Testosterone was higher in CW (Median 23 ng/dl, min-max 10-37) than TW (Median 12 ng/dl, min-max 10-35). Significant associations found in TW between QoL and the measured hormones were: E2/Vitality (Rs -0,32, p 0,03). and LH/Physical role (Rs 0,33, p 0,02). None was found in CW. CONCLUSION: TW and CW had similar scores in  QoL.  Hormone profile of TS patients, when treated, slightly likens to the woman in her follicular stage.  Oestradiol is directly related to QoL, and so is LH, highlighting the importance of the Hormonal Replacement therapy to keep a good QoL.


Nothing to Disclose: MBE, PTM, KCO, ITNV

PP10-3 25794 3.0000 FRI 144 A Do Concentrations of Sex Hormones Affect Quality of Life in Turner Syndrome? 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Reproductive Endocrinology Friday, April 1st 11:45:00 AM PP10 7725 11:30:00 AM Recent Developments in Disorders of Sex Development (DSD) and Transgender Medicine Poster Preview

Andy Videsh Babwah*1, Silvia Leon2, Michele Dawn Calder1, Moshmi Bhattacharya3, Kanako Hayashi4, Stephen Power3, George A Vilos3, Angelos G Vilos3 and Manuel Tena-Sempere5
1University of Western Ontario, London, ON, Canada, 2University of Cordoba, Cordoba, Spain, 3University of Western Ontario, 4Southern Illinois University, Carbondale, IL, 5University of Cordoba, Instituto Maimonides de Investigacion Biomedica de Cordoba (IMIBIC/HURS), CIBERobn Instituto Carlos III, Cordoba, Spain


Kisspeptins (KPs) are a group of related peptides that signal via KISS1R and in the brain potently trigger GnRH secretion and thereby act as a central regulator of reproduction. Recent studies from the Tena-Sempere and Babwah laboratories provide compelling evidence that peripherally-derived KP also regulates reproduction at the level of the ovary and uterus (1; 2). During our studies, we observed that the uteri from adult global Kiss1-/- and Kiss1r-/- (KO) mice contain significantly fewer endometrial glands compared to their WT littermates (2). Endometrial glands are found in all mammalian uteri and glands produce substances that are essential for the establishment of uterine receptivity and embryo implantation and survival. Thus, mutations that diminish or ablate endometrial gland formation result in subfertility or infertility. In many species including rodents, gland development (adenogenesis) occurs in a steroid-independent manner in the neonatal and juvenile periods but as ovarian hormones increase with the onset of puberty, adenogenesis shifts into an ovarian-dependent phase that lasts throughout adult life. An ovarian hormone strongly suggested to regulate adenogenesis is estradiol (E2).  However, except for the preovulatory E2 surge, E2 levels are similar in adult KO female mice compared to WT littermates, a finding supported by the observation that follicles develop to the pre-antral stage, thus, a source of E2 exists in these mice. Together, these observations strongly suggest that E2 is unlikely the limiting factor that underlies reduced adenogenesis in these mice. To investigate the cause of reduced adenogenesis in adult KO mice we are comparing gland number between juvenile (3-wk-old) Kiss1-/- mice and WT littermates. Preliminary findings reveal that juvenile Kiss1-/- mice contain 30% fewer glands than WT littermates suggesting that the reduced adenogenesis observed in adults is due to an absence of factors that are lacking in the neonatal and juvenile periods. Such factors might be independent of the ovary and instead reflect a lack of uterine kisspeptin signaling. As a complementary study, we will compare gland number between adult Kiss1r-/-Tg mice [these are Kiss1r-/- mice in which Kiss1r is specifically re-expressed in the GnRH neuron leading to an activation of the hypothalamic-pituitary-gonadal axis (3)] and Kiss1r-/- and WT littermates. If Kiss1r-/-Tg and Kiss1r-/- mice have similar gland numbers, it would suggest this is not due to ovarian-derived factors (but maybe lack of kisspeptin signaling), whereas if gland number is higher in Kiss1r-/-Tg than Kiss1r-/- mice, this would be compatible with a role of ovarian-derived factors. However, since E2 levels are similar between adult Kiss1r-/- and WT mice it is unlikely that this factor is E2.  


Nothing to Disclose: AVB, SL, MDC, MB, KH, SP, GAV, AGV, MT

PP10-4 25179 4.0000 FRI 154 A Investigating the Cause of Reduced Adenogenesis in Adult Kiss1-/- and Kiss1r-/- mice 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Reproductive Endocrinology Friday, April 1st 11:45:00 AM PP10 7725 11:30:00 AM Recent Developments in Disorders of Sex Development (DSD) and Transgender Medicine Poster Preview

Genevieve V Dall*1, Jessica L Vieusseux2, Kenneth S Korach3, Yukitomo Arao3, Sylvia C Hewitt3, Katherine J Hamilton3, Wah Chin Boon4, Evan R Simpson5, Robert G Ramsay2, Robin L Anderson2, Gail P Risbridger1 and Kara L Britt2
1Monash University, Melbourne, Australia, 2Peter MacCallum Cancer Centre, Melbourne, Australia, 3National Institute of Environmental Health Sciences/NIH, Research Triangle Park, NC, 4The Florey Institute of Neuroscience and Mental Health, Parkville VIC, Australia, 5MIMR-PHI Institute of Medical Research, Clayton VIC, Australia


Estrogen stimulates breast development during puberty and mammary tumours in adulthood through the estrogen receptor α (ERα). These effects are believed to occur via ERα+ luminal mature cells and not the mammary stem cells (MaSC) which are ERα-ve. The ERα+ve luminal mature cells express the stem cell antigen 1 (Sca-1), and this has recently been used to further define an ERα+ve subset of luminal progenitors cells. Considering the estrogen sensitivity of mammary stem cells, we sought to determine if Sca-1 might indeed define an ERα+ population within the CD24+ CD49fhi MaSC-enriched population. Lineage negative CD24+ CD49fhi MaSC cells exhibited a distinct Sca-1+ve population that was abundant in pre-pubertal mammary glands which decreased in adulthood. Similar to their Sca-1-ve counterparts, they were quiescent pre-pubertal, but importantly were the only cells to enter the cell cycle at puberty suggesting they possessed hormone receptors. We showed here that whilst they did not contain high levels of MaSC markers, nor high in vivo transplantation activity, they expressed stem cell markers, and had limited multi-potent stem cell activity. We assessed ERα expression and found transcript and protein as well as expression of the ERα target gene, progesterone receptor. As Sca-1+ MaSC decreased upon hormonal stimulation associated with puberty, we assessed whether they were also estrogen sensitive. The mammary glands of estrogen-deficient aromatase knockout (ArKO) and ERa deficient (αERKO) mice (with characteristic rudimentary mammary glands) possessed only Sca-1+ MaSC and completely lacked Sca-1-ve cells. This phenotype was rescued, if we treated ArKO mice with estrogen pellets from postnatal day 5 where ductal growth was induced and Sca-1-ve MaSC were restored. In conclusion, Sca-1 enriches for an ERα+ve, estrogen-sensitive subpopulation within the CD24+ CD49fhi MaSC population that are likely to be responsible for the hormonal sensitivity of the developing mammary gland.


Nothing to Disclose: GVD, JLV, KSK, YA, SCH, KJH, WCB, ERS, RGR, RLA, GPR, KLB

PP06-1 25382 1.0000 FRI 206 A Sca-1 Delineates an Estrogen Responsive Stem Cell within the Mammary Gland 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Nuclear Receptors and Steroid Hormone Action Friday, April 1st 11:45:00 AM PP06 7736 11:30:00 AM Nuclear Receptors and Co-Regulators in Health and Disease Poster Preview

Charles E Foulds*1, Leah A Gates1, Ross A Hamilton1, Philip F Lavere1, Ping Yi1, Yang Yu1, David M Lonard1, Jun Qin1, Anna Malovannaya1, Guowei Gu1, Suzanne A Fuqua1, Jieya Shao2, Matthew J Ellis1 and Bert W O'Malley1
1Baylor College of Medicine, Houston, TX, 2Washington University School of Medicine, St. Louis, MO


Approximately 70% of breast cancers are estrogen receptor-α (ERα)-positive and are driven by estrogen. Targeting ERα with endocrine therapies, such as tamoxifen and aromatase inhibitors, provides effective adjuvant treatment for this patient subset with reductions in recurrence approaching 50%. Still, many tumors acquire resistance, and in ~20% of these cases, gain-of-function mutations in the ERα gene (ESR1) are implicated. Amino acid substitutions in the receptor’s ligand binding domain (LBD) result in mutant receptors displaying ligand-independent activity, enhanced binding to some steroid receptor coactivators (SRCs), and resistance to subsequent endocrine therapy (e.g., anti-estrogen fulvestrant). In addition, rarer translocations of ESR1 to other genes create fusion proteins that lack the LBD and cannot be targeted with anti-estrogens. Therefore, new approaches are critically needed in treating ERα-positive metastatic breast cancer in patients expressing these different ERα mutants. Our studies focus on the most prominent of the ERα LBD mutants, Y537S and D538G, and the ESR1-YAP1 fusion protein. While efforts to develop more effective anti-estrogens (e.g. new oral Selective ER Downregulators, SERDs) may be useful, additional therapeutic strategies designed to concomitantly target coactivators (CoAs) offer a unique approach to enhance treatment (or prevent development) of recurrent disease. To accomplish targeting of the proper CoA, the set of CoAs that bind mutant ERα proteins must first be determined. Here, we have setup a mass spectrometric, unbiased proteomic approach, based on our prior published “estrogen response element (ERE) DNA pulldown” system with wild-type (WT) ERα (1), to profile the entire CoA “complexome” for each mutant ERα protein. Mass spectrometric data of CoA recruitment to each mutant was compared to the WT receptor to identify potential new therapeutic targets. Interestingly, we observed specific CoAs (or complexes) indeed display robust enhanced binding to a mutant ERα protein over the WT receptor (e.g., SRCs, p300, MLL4 complex with Y537S; SRC-1, SRC-3, p300, and SAGA complex with D538G; 26S proteasome with ESR1-YAP1 fusion protein). Based on these findings, we tested the effect of inhibiting select CoA candidates on their ability to activate transcription of an ERE-driven luciferase reporter mediated by the different ERα mutant receptors. We found that a new “pan-SRC” small molecule inhibitor (SMI) called SI-1 reduced Y537S and D538G activities, the proteasome inhibitor MG-132 reduced ESR1-YAP1 activity, and siRNA targeting MLL4 reduced Y537S activity. These data suggest that SMIs targeting these distinct CoAs may be promising new therapeutics, as single agents or in combination with new oral SERDs, to inhibit growth of breast cancer cells expressing these ERα mutant proteins.


Disclosure: MJE: Consultant, Astra Zeneca. Nothing to Disclose: CEF, LAG, RAH, PFL, PY, YY, DML, JQ, AM, GG, SAF, JS, BWO

PP06-2 27001 2.0000 FRI 212 A Different Mechanisms Utilized By Mutant Estrogen Receptor Alpha Proteins to Activate Transcription in a Ligand-Independent Manner 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Nuclear Receptors and Steroid Hormone Action Friday, April 1st 11:45:00 AM PP06 7736 11:30:00 AM Nuclear Receptors and Co-Regulators in Health and Disease Poster Preview

Shino Murakami*, Anusha Nagari and W Kraus
University of Texas Southwestern Medical Center, Dallas, TX


Estrogen receptor alpha (ERa) is a ligand-regulated transcription factor that nucleates the formation of enhancers across the genome in response to estrogen.  ERa enhancers have all of the features that are associated with active enhancers (e.g., H3K4me1, H3K27ac, enrichment of p300/CBP and Mediator, and the enhancer RNA production), although the roles these features play in ERa enhancer function are not well understood.  p300 and CBP are protein acetyltransferases that are recruited to liganded ERa at enhancers through the steroid receptor coactivators (SRCs; a.k.a. p160 coregulators), which interact directly with liganded ERa.  Mediator, a multisubunit coregulator complex that is thought to bridge enhancers and the core transcription machinery, also interacts directly with ERa through the Med1 subunit.  Interestingly, SRC proteins and Med1 interact in a mutually exclusive manner with the same ligand-induced hydrophobic cleft on ERa.  We are using a series of ERa mutants in combination with biochemical, molecular, genomic, genetic, and chemical approaches to dissect the mechanisms of assembly and functions of ERa enhancers.  A mutant ERa that selectively binds Mediator versus SRCs promotes the formation of enhancer-promoter chromatin loops, but fails to recruit RNA polymerase II at the promoters and enhancers, suggesting that looping is not sufficient to activate gene expression.  This mutant ERa activates transcription at the enhancers and promoters at early time points in the estrogen response, but does not sustain active transcription, suggesting that SRCs are required to maintain an active enhancer.  Additional genomic and molecular approaches are providing insights into ERa enhancer complex assembly, chromatin regulation, enhancer-promoter looping, and transcription activation.


Nothing to Disclose: SM, AN, WK

PP06-3 27033 3.0000 FRI 210 A Molecular Mechanisms for the Assembly and Function of Estrogen Receptor Enhancers 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Nuclear Receptors and Steroid Hormone Action Friday, April 1st 11:45:00 AM PP06 7736 11:30:00 AM Nuclear Receptors and Co-Regulators in Health and Disease Poster Preview

J Dinny Graham1, Heidi Nicole Hilton2, Audrey Silvestri1, Nicole Santucci3, Tram B Doan1, Lily Huschtscha4 and Christine L Clarke*1
1University of Sydney, Westmead Millennium Institute, Westmead NSW, Australia, 2University of Sydney, Westmead Millennium Institute, Sydney, Australia, 3University of Sydney, Westmead Millennium Institute, Westmead, Australia, 4Children's Medical Research Institute, Westmead, Australia


The ovarian hormone progesterone (P) is a key regulator of female reproductive function. It plays an essential role in the development of lobular alveolar structures in the breast, through stimulation of proliferation during the normal menstrual cycle and pregnancy. This contrasts with its role in the uterus, suppressing prostaglandin signalling to maintain pregnancy and opposing the proliferative effects of estrogen (E) in the endometrium. The established role of P analogues in hormone replacement therapy in increasing breast cancer risk has sharpened focus on the mechanisms of action of this hormone in the normal breast. The finding that progestins in hormone replacement therapy (HRT) increased breast cancer risk was completely unexpected, as was the lack of any increased risk associated with E in HRT. The dogma that P is an anti-estrogen in the breast is challenged by the findings of the HRT trials; and by emerging evidence from our group and others that P is proliferative in the normal human breast; that P expands progenitor cells in human breast and mouse mammary gland; and that P augments cancer stem cells. Also, in a primary cell culture model of normal human breast we revealed that PR and ER are functionally distinct in the normal breast: PR was present in cell populations enriched for uncommitted progenitors, whereas ER was detected in luminal progenitors. Moreover, their detection in breast tissue by dual immunofluorescence revealed striking heterogeneity and non-overlapping expression in mature cells. We have now extended our findings to single cell resolution. The normal mixed lineage structures that formed on 3D culture of primary normal breast organoids were harvested, dissociated to single cells and individual cells were captured for gene expression profiling. Principal component analysis of lineage and proliferation marker expression revealed three distinct cell populations: one with predominantly luminal characteristics, one predominantly basal and a bipotent population expressing both luminal and basal markers and high in stem/progenitor markers. While the single cell profiling data broadly supported current understanding of the epithelial hierarchy, there were some striking findings. Firstly, a significant number of PR+ cells were basal cells and/or progenitors. Moreover, in basal cells, PR expression tracked with high expression of stem/progenitor markers, and proliferation genes. Secondly, supporting our previous findings, PR and ER expression was largely non-overlapping. Taken together, our data suggest distinct roles for ER and PR in the regulation of proliferation and lineage progression in the normal breast and provide an explanation for the different carcinogenic potential of E and P in the development of breast cancer.


Nothing to Disclose: JDG, HNH, AS, NS, TBD, LH, CLC

PP06-4 27405 4.0000 FRI 205 A Single Cell Analysis Reveals Divergent Progesterone and Estrogen Signaling Potential in the Normal Human Breast 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Nuclear Receptors and Steroid Hormone Action Friday, April 1st 11:45:00 AM PP06 7736 11:30:00 AM Nuclear Receptors and Co-Regulators in Health and Disease Poster Preview

Luiz Guilhemme Grossi Porto*1, Maria N Korre2, Steven Moffatt3 and Stefanos N Kales2
1Harvard T. H. Chan School of Public Health and University of Brasilia Faculty of Physical Education, Brazil, Boston, MA, 2Harvard T. H. Chan School of Public Health and Cambridge Health Alliance, Harvard Medical School, Boston, MA, 3Public Safety Medical, Indianapolis, IN


Impaired heart rate recovery (HRR) after exercise testing and obesity have been described as risk factors for cardiovascular diseases and overall mortality. The obesity paradox phenomenon has shown that physical fitness could be an important modifier effect when obesity is associated with higher cardiovascular and mortality risks. We aimed to evaluate the associations between obesity, physical fitness and heart rate recovery after exercise testing among firefighters. We performed a cross sectional study within a large US fire department, applying a weighted sampling strategy, selecting randomly 100 participants from the eligible population; 75 at low cardiovascular risk and 225 at high risk. Inclusion criteria were: male > 18 yrs old with a recorded submaximal Bruce-treadmill test and no restrictions on duty. Impaired HRR was defined as HRR < 18 bpm at the 1st or < 42bpm at the 2nd min of the recovery period after exercise testing. Cardiorespiratory fitness (CRF) was estimated by the treadmill test and categorized as low and high. Muscle strength was evaluated by the push-ups test (PUT) and obesity was defined as BMI ≥ 30 kg/m2. 30 participants were excluded due to incomplete data. Crude associations were estimated by the odds ratio (OR-95%CI) and the multivariate analysis was assessed by binary logistic regression. Mann-Whitney test was used when applicable, at the 5% level of significance. Participants were 46.5 ± 8.3 years old, with BMI equal to 31.1 ± 4.7 km/m2. Prevalence estimates were 21.5% for HRR impairment and 55.4% for obesity. Median (interquartile range) of HRR at the 1st and 2nd min were 37 (27 - 46) and 54 (44 - 62) bpm, respectively. The odds of having impaired HRR were 2.74 (1.58 - 4.75) among obese as compared to non-obese; 3.11 (1.43 - 6.76) for those ≥40 years old vs the youngers and 2.71 (1.61 - 4.56) for those with lower CRF vs the fittest ones. Participants with normal HRR had better PUT than those with impaired HRR [25 (16 - 30) vs 16.5 (18 - 10), respectively)(p<0.001). After adjustments for age, push-ups, BMI and CRF as categories, only the number of push-ups remained significantly associated with impaired HRR, with OR=0.96 (0.94 - 0.99).  Physical fitness appears to mediate age and obesity related HRR impairment. Each extra push-up was associated with 4% reduction in the odds of having reduced HRR profile. Our data support the need for considering physical fitness level in cardiovascular risk analyses among firefighters and to recommend muscle training as a potential cardiovascular protection factor.


Nothing to Disclose: LGGP, MNK, SM, SNK

PP07-2 26947 2.0000 FRI 605 A Muscle Strength Is a Better Predictor of Impaired Heart Rate Recovery Than BMI Among Firefighters 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Obesity, Adipocyte Biology and Appetite Friday, April 1st 11:45:00 AM PP07 7744 11:30:00 AM Predictors of Weight Gain and Disease Poster Preview

Corey J Lager*1, Nazanene H Esfandiari1, Andrew T Kraftson2, Angela R Subauste3, Amy L Lockwood1, Darlene Bellers1, Morton B Brown1, Oliver A Varban1 and Elif A Oral1
1Univ of Michigan, Ann Arbor, MI, 2University of Michigan, Ann Arbor, MI, 3University of Mississippi Medical Center, Jackson, MS


Background: Bariatric surgery appears to be more effective than medical treatment for weight loss. It remains unclear if surgery is capable of achieving normal weight and sustaining it long term for all patients.  The annual probability of attaining normal weight was recently estimated to be 1 in 1,290 for males and 1 in 677 for females with morbid obesity using medical interventions after age 20.

Hypothesis: Gastric bypass surgery will result in sustained weight loss over time with an increased likelihood of weight normalization compared to published rates for medical therapy.

Methods: We carried out retrospective chart review of patients undergoing gastric bypass surgery (GB) at the University of Michigan between January 1, 2008 and November 1, 2010 who also followed up in the post bariatric surgery clinic (n=219). We evaluated the sustainability of weight loss and the likelihood of achieving a normal weight (BMI<25 kg/m2) 5 years after gastric bypass surgery.

Results: There were 183 females (age: 43±11 years, BMI 47.0±7.9 kg/m2) and 36 males (age: 48±9 years, BMI 46.9±7.6 kg/m2). Out of the female patients, 123 had data at 2 years and had lost a mean of 72.7±25.7% of their excess weight. Four and 5-year follow up data are available on 112 and 105 patients, and mean % excess weight lost from baseline was 61.6±24.5 and 59.2±25.8 % respectively. For males, 2, 4 and 5-year data was available on 22, 22 and 18 patients, and mean % excess weight lost from baseline was 55.2±41.3, 39.6±64.9, and 29.7±72.5% respectively. The difference between the two sexes did not attain statistical significance (p=0.06, 0.07, 0,12 respectively), but males displayed greater variability in the amount of weight loss compared to females. The number of female patients achieving a BMI of 25 kg/m2 or better during the 5 years of observation was 31 (16.9 %) while only 5 of these patients reached normal weight by year 2 and sustained it for more than 3 years.  There were only 2 males (5.5%) who reached a BMI of 25 during the 5-year follow-up. From the females, only 13 patients out of 105 who returned for follow up regained more than half of the weight they lost while none regained all the weight they lost. In males, 4 of the 18 patients returning regained more than half of the weight they lost and 2 experienced regain of all weight lost.

Conclusions: Our data adds to the growing body of evidence that GB provides sustainable weight loss in patients with morbid obesity who return for follow-up. Although the percentage of patients who reached and sustained normal weight is greater than what is reported with medical weight loss, this rate was still low, underscoring the challenge of weight normalization in patients with a very high initial body mass index. Further studies are required to determine preoperative markers that can predict long-term success with bariatric surgery and whether weight normalization can be an attainable goal for specific subgroups of patients.


Disclosure: ATK: Clinical Researcher, Optifast (Nestle, PA). EAO: Principal Investigator, Isis Pharmaceuticals, Principal Investigator, Aegerion Pharmaceuticals, Principal Investigator, GI Dynamics, Advisory Group Member, Astra Zeneca, Principal Investigator, Astra Zeneca. Nothing to Disclose: CJL, NHE, ARS, ALL, DB, MBB, OAV

PP07-3 26179 3.0000 FRI 637 A Likelihood of Achieving Normal Weight 5 Years after Gastric Bypass Surgery: Results from the University of Michigan Post Bariatric Surgery Clinic 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Obesity, Adipocyte Biology and Appetite Friday, April 1st 11:45:00 AM PP07 7744 11:30:00 AM Predictors of Weight Gain and Disease Poster Preview

Olivia M. Farr*1, Jagriti Upadhyay2, Anna Gavrieli3, Hannah Mathew3, Maria T Vamvini4, Michelle Camp3, Harper Kaye3, Nikolaos Spyrou3, Anastasia Koniaris3, Holly Kilim3, Alexandra Srnka3, Alexandra Migdal5 and Christos S. Mantzoros6
1Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, 2Harvard Medical School/ Beth Israel Deaconess Medical Center, Jamaica Plain, MA, 3Harvard Medical School/ Beth Israel Deaconess Medical Center, 4Mount Auburn Hospital, Cambridge, MA, 5Beth Israel Deaconess Medical Center, Boston, MA, 6BIDMC, Harvard, Boston, MA


Lorcaserin is a 5HT-2c receptor agonist effective in treating obesity. While studies in rodents have shown that lorcaserin acts in the brain to exert its weight reducing effects, this has not yet been confirmed in humans. Also, which brain centers may be activated in humans remains unknown. We performed a randomized, placebo-controlled, double-blind trial with 48 obese participants to study the effects of lorcaserin on the brain using functional magnetic resonance imaging (fMRI) after 1 week and 4 weeks of therapy. We found decreased brain activations in the parietal and visual cortices in response to highly palatable food cues at 1 week in the fasting state and in the parietal cortex in response to any food cues at 4 weeks after a standardized meal. Decreases in weight and caloric intake correlated with activations in parietal and visual cortices to highly desirable food cues at baseline. Additionally, in a whole brain regression analysis, we observed that activations of amygdala to highly desirable food cues at baseline correlated to weight lost at 4 weeks. Altogether, this data suggests that lorcaserin exerts its weight reducing effects by decreasing attention-related brain activations to food cues, including the parietal and visual cortices and/or amygdala. Results from the regression analysis suggest that lorcaserin is of particular benefit to individuals who are emotional eaters. These data need to be confirmed and extended by future studies.


Nothing to Disclose: OMF, JU, AG, HM, MTV, MC, HK, NS, AK, HK, AS, AM, CSM

PP07-4 24157 4.0000 FRI 600 A Lorcaserin Decreases Activation of Attention-Related Brain Centers in Response to Food Cues during a Four-Week-Long Randomized, Placebo-Controlled, Double-Blinded Clinical Trial 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Obesity, Adipocyte Biology and Appetite Friday, April 1st 11:45:00 AM PP07 7744 11:30:00 AM Predictors of Weight Gain and Disease Poster Preview

Allison R. Smego*, Philippe Backeljauw and Iris Gutmark-Little
Cincinnati Children's Hospital Medical Center, Cincinnati, OH


Background: The treatment of neurogenic diabetes insipidus (DI) in infancy is challenging and often associated with wide fluctuations between fluid overload and dehydration. Therapy with subcutaneous, intranasal (IN), or oral tablet desmopressin acetate (DDAVP) is difficult to titrate in infants. Use of thiazide diuretics and low renal solute load formulas, or administration of extra free water, all may impair caloric intake and are not conducive to a stable sodium and fluid balance.

 Objective: Assess the efficacy and safety of orally administered IN DDAVP for the management of infants with neurogenic DI.

 Design and methods: Retrospective review of clinical and laboratory data of fifteen infants (mean age: 4.5 months) with neurogenic DI treated at a tertiary care center. Treatment was with diluted IN DDAVP formulation (10 mcg/mL) administered orally via a tuberculin syringe to the buccal mucosa.

 Results: After initial DDAVP titration over 2-3 days, IN DDAVP doses ranged from 1 μg to 5 μg twice daily given orally. Mean sodium concentration at DI diagnosis was 159±6.6 mmol/L (range, 151-178) and improved to 142±3.5 mmol/L (range, 137-147) with the orally administered IN DDAVP. Normal serum sodium concentrations were achieved without major fluctuations. Serum sodium was then maintained in the outpatient setting at a mean of 146±5.2 mmol/L (mean duration of follow-up: 4 months).

 Conclusions: Orally administered IN formulation of DDAVP provides a practical and safe treatment alternative for neurogenic DI in infancy. Our experience was most notable for avoidance of severe hypo- and hypernatremia during DDAVP titration and ongoing outpatient management of DI. The possibility for smaller dosage increments and ease of administration make IN DDAVP administered orally preferable over other DDAVP treatment options in infants.


Disclosure: PB: Advisory Group Member, EMD Serono, Advisory Group Member, Sandoz, Advisory Group Member, Novo Nordisk, Advisory Group Member, Ipsen. Nothing to Disclose: ARS, IG

PP08-1 26131 1.0000 FRI 049 A Efficacy and Safety of Orally Administered Intranasal Formulation of Desmopressin Acetate for the Management of Neurogenic Diabetes Insipidus in Infancy 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Pediatric Endocrinology Friday, April 1st 11:45:00 AM PP08 7759 11:30:00 AM Pediatric Endocrinology Poster Preview

Mirela Costa de Miranda*1, Eliane Pereira dos Santos2, Daniel Fiordelisio de Carvalho1, Andresa De Santi Rodrigues3, Ivana Van Der Linden Nader2, João Amélio da Silva Junior2, Berenice B Mendonca3 and Tania A Bachega1
1Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brazil, 2ASSOCIAÇÃO DE PAIS E AMIGOS DOS EXCEPCIONAIS (APAE) do Estado de Goiás, 3Laboratório de Hormônios e Genética Molecular- LIM/42, Unidade de Adrenal, Disc. de Endocrinologia e Metabologia, Hospital das Clínicas, Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brazil


Introduction: Congenital adrenal hyperplasia (CAH) presents the criteria for inclusion in newborn screening (NBS) programs, which is highly effective in indentifying the severe cases. However, the high rate of false-positive results (FPR) remains an important issue. Therefore, positive neonatal tests must be confirmed by serum 17OHP levels, which present a great overlap among cases with SW, SV, NC forms and those with FPR, leading to therapeutic implications. Stressed newborns could present persistently increased hormonal confirmatory tests, requiring, sometimes, prolonged follow-up.

Objective: To evaluate the utility of molecular analysis to improve CAH diagnosis in our NBS program.

Material and Methods: N17OHP were measured by IFMA assays (Autodelfia-Perkin Elmer) and cutoffs adjusted for birth-weight. Confirmatory tests included serum 17OHP, androstenedione, testosterone and cortisol analyses. DNA were extracted from 70 newborns with increased neonatal (N) and serum 17OHP levels, between 1999-2014. CYP21A2 genotypes were determined by allele-specific PCR and MLPA techniques; entire gene sequencing was performed when necessary.

Results: Among all newborns with positive tests, 40 (57%) presented genotypes predicting classical forms (21 males); 33/40 (83%) presented the SW form. Regarding patients carrying the I2 splice as the less severe affected allele (22), 16 (73%) presented slightly or moderate hyponatremia in the first days of life; in the 6 remaining, the clinical form distinction was not possible, due to the precocious treatment introduction.  All patients carrying the p.I172N mutation, in the less affected allele, had the SV form. Genotyping identified mutations in 100% of classical alleles. Among the 30 asymptomatic newborns (15 males) with persistently increased serum 17OHP levels, genotyping predicted NC form in 15 (21.5%) and the other 15 with non-affected genotypes (including 6 heterozygotes) were discharged. Mean N17OHP in classical patients (confirmed by genotyping) was 327 (±163) ng/mL, whereas in the others, including FPR and nonclassical newborns, was 137 (±128) ng/mL. In affected newborns, mutations derived from gene conversion events were found in 89% of the alleles; the most frequent were I2 splice (35%), p.Q318X (23%) and p.R356W (19% of alleles). Mutations not derived from pseudogene were found in 11% of the alleles: gene founder effect was observed with the p.G424S, p.R408C and IVS2-2A>G mutations.

Conclusion: molecular testing was a useful supplemental tool especially in identifying false-positive results in CAH-NBS, preventing unnecessary follow-up of cases with inconclusive hormonal tests. We observed a good genotype/phenotype correlation being helpful to predict the clinical forms in asymptomatic affected newborns. Additionally, molecular diagnosis in our population should take into account mutations not derived from pseudogene.



PP08-3 26767 3.0000 FRI 011 A Molecular Confirmatory Test Improves the Accuracy of Congenital Adrenal Hyperplasia Diagnosis in Newborn Screening Program 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Pediatric Endocrinology Friday, April 1st 11:45:00 AM PP08 7759 11:30:00 AM Pediatric Endocrinology Poster Preview

Yuta Chiba*, Daisuke Sugawara, Yasuko Tanaka, Yumiko Terada, Yasuhiro Naiki and Reiko Horikawa
National Center for Child Health and Development, Tokyo, Japan


Background: Recent increase of childhood and adolescent obesity leads to the increase of fatty liver change and nonalcoholic fatty liver disease (NAFLD) in young age. Although liver biopsy is the gold standard for the diagnosis of NAFLD, non-invasive biomarkers and images may reflect this condition. P IIIP is known to be one of the biomarkers for liver fibrosis. In this study, we evaluate P IIIP levels as a marker of fatty and fibrotic changes of liver in obese children.

Subjects and Methods: Ninety-five children with increasing weight (male:67) aged median 10.0 (5.0 to 14.5) were involved in this study. Mean percent obesity was +43.6%(+12.8~+113.8%,<20%:5,20~30%:13,30~50%:49,50%<:28). Serum P IIIP levels and liver function were measured in all subjects. Abdominal CT or ultrasonography were performed in 78 cases.

Results: 24% of the patients had elevated AST/ALT/γGTP levels. 39 out of 78 are diagnosed to have fatty liver/mild fatty changes. Serum PIIIP levels were ranged from 0.65 to 3.0 U/ml (reference range: 0.3~0.8). There was no significant difference in P IIIP levels between patients with or without fatty liver diagnosed by imaging (1.17±0.30 (mean±SD) and 1.18±0.45 U/ml, respectively). However, there was significant difference of P IIIP levels in two groups with or without obesity (0.83±0.18 and 1.16±0.37 U/ml, respectively, ; P = 0.01). There was significant positive correlation between P IIIP levels with subcutaneous fat amount.

Discussion: P IIIP levels were significantly higher in patients with obesity. P IIIP levels did not significantly differ in patients with/without fatty liver change assessed by CT or ultrasonography. It may be due to the lower sensitivity of images, or P IIIP levels reflecting fibrotic changes of liver rather than fatty changes and no complete concordance of these two conditions. 

Conclusion: Serum P IIIP levels were elevated with progression of obesity, indicating that liver fibrosis with fatty changes can occur and progress in children and adolescents. Serum P IIIP measurement may be a useful tool to detect early pathologic changes of liver.


Nothing to Disclose: YC, DS, YT, YT, YN, RH

PP08-4 25550 4.0000 FRI 001 A The Evaluation of Procollagen Type III N-Terminal Peptide(P IIIP) As a Marker of Fatty Liver Change in Obese Children and Adolescents 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Pediatric Endocrinology Friday, April 1st 11:45:00 AM PP08 7759 11:30:00 AM Pediatric Endocrinology Poster Preview

Thozhukat Sathyapalan*1, Lesa Aylward2, Natalie J Thatcher3, Martin Rose4, Steve Petch5, Alwyn Fernandes5 and Stephen L Atkin6
1Michael White Diab Cntr, Scotland, United Kingdom, 2Summit Toxicology, LLP, Falls Church, VA, USA, 3European Food Safety Authority, Parma, Italy, London, Italy, 4FERA, Sand Hutton, York, United Kingdom, United Kingdom, 5FERA, Sand Hutton, York, United Kingdom, 6Weill Cornell Medical College Qatar, Doha, Qatar



There is increasing concern about the endocrine disrupting chemicals and their involvement in obesity, diabetes, fertility and cancer. However, much baseline data is missing and many of these chemicals are lipophilic and sequester in fat; therefore as the prevalence of obesity is increasing in most populations this study was performed.  The aim of this study was to provide baseline data on the concentrations of chlorinated and brominated dioxins and related compounds as well as polybrominated diphenyl ethers to assess whether concentrations of these compounds are higher in obese than control subjects.

Materials and Methods

Patients undergoing Roux-en-y gastric bypass surgery for weight loss and control patients who were undergoing abdominal surgery for non-bariatric reasons were recruited with informed consent for the study.  Anthropometric parameters were measured at the day of surgery.  During surgery, visceral and subcutaneous adipose tissue biopsies, liver biopsy and blood samples were taken.


Patients undergoing bariatric surgery were younger on average than control patients (47.9 (12.7) vs. 68.5 (14.2) years) and on average had higher BMI (47.1 (10.8) vs. 25.3 (4.9) kg/m2). 

Tissue concentrations were measured in samples of visceral and subcutaneous fat and in liver biopsies. Since the bariatric and control surgery groups were significantly different in age, had overlapping body mass index (BMI) ranges, and due to the known age-dependent patterns of tissue concentrations for chlorinated toxic equivalency (TEQ), the data from the two groups were combined for further analysis. Brominated TEQ concentrations were relatively low compared to chlorinated TEQ, constituting less than 5% of adipose tissue TEQ and less than 10% of liver TEQ.  The most frequently detected PBDD/F compounds were 2,3,7,8-tetrabromodibenzodioxin, 2,3,7,8-tetrabromodibenzofuran, and 2,3,4,7,8-pentabromodibenzofuran. The PBDE compounds presented here are those that were consistently detected in the samples.  Of these, BDE 153 was present at the highest concentrations, followed by BDE 47.

Multivariate linear regressions showed chlorinated TEQ in visceral fat was significantly positively associated with both age and BMI.  In contrast, brominated TEQ compounds showed no significant association with any of the factors considered.  BDE 47 was borderline significantly negatively associated with age, while BDE 153 showed a borderline significant negative relationship to BMI.  Gender was not a significant factor for any analyte. 


Subcutaneous fat concentrations were highly correlated with visceral fat concentrations for all analytes. This confirms that concentrations of these compounds in fat depots in the body appear to be generally in equilibrium, an observation previously made for chlorinated TEQ compounds but not previously demonstrated in humans for PBDD/Fs and PBDEs.


Nothing to Disclose: TS, LA, NJT, MR, SP, AF, SLA

PP04-1 25325 1.0000 FRI 106 A Concentrations of Chlorinated and Brominated Dioxins, Furans and PBDEs in Human Liver, Subcutaneous and Visceral Adipose Tissue in the United Kingdom 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Endocrine Disruptors Friday, April 1st 11:45:00 AM PP04 7763 11:30:00 AM Endocrine Disrupting Chemicals Poster Preview

Rebekah C Kennedy*1, Russell R Fling1, Michael Robeson2, Arnold M Saxton1, David Bemis1, Jiang Liu1, Ling Zhao1 and Jiangang Chen1
1University of Tennessee, Knoxville, TN, 2Colorado State University, Fort Collins, CO


The use of prescription antibiotics during pregnancy and birth can induce alterations in the maternal bacterial milieu and contribute to the disturbance of neonatal microbial colonization. The assumed risk of infectious disease during pregnancy and lactation additionally leads to widespread use of non-prescription antimicrobials in household products. Triclocarban (3,4,4′-trichlorocarbanilide; TCC) is an antimicrobial compound added to bar soaps for its bacteriostatic properties. We previously demonstrated that TCC concentrates in and is transferred through the milk to suckling neonatal rats, potentially leading to alterations in the diversity of the gut microbiota. To date, the consequence of TCC exposure during early life on gut microbial composition is unknown.      

Timed-pregnant SD rats were provided ad lib access to TCC supplemented diet (0.1% w/w) starting at gestational day (GD) 4 until postnatal day (PND) 16 after birth. Fecal samples were collected from dams during gestation and lactation. Cecum content was collected from neonates during lactation only. A group of age-matched unexposed dams and their neonates served as controls. The V4 region of the 16S rRNA region was sequenced via the MiSeq platform.  Sequences were analyzed with the Phyloseq and Vegan packages in R.

Exposure to TCC during gestation and lactation led to perturbations in microbial community structure across time in both dams and neonates. Phylogenetic diversity was significantly reduced among exposed dams from GD 11 until sacrifice and neonates at PNDs 12 and 16. Weighted Unifrac analysis of microbiota from TCC exposed dams revealed significant dysbiosis of the gut microbiota by GD 18, a trend that continued to 16 days after delivery when the samples were last collected. Using the same metric among neonates, in both control and TCC exposed animals, an initial stochastic pattern emerges at PND 3. At PND 6, an overall restructuring occurs where both control and TCC exposed communities converge. By PND 12, communities start separating based on exposure status and become significantly different at PND 16.

Our results both demonstrate the impact of TCC exposure on gut microbial structure during gestation and lactation, as well as provide insight into the neonatal bacterial colonization process. The ability of TCC to drive microbial dysbiosis warrants future investigation to determine the clinical health outcomes resulting from non-prescription antimicrobial use during sensitive exposure windows.


Nothing to Disclose: RCK, RRF, MR, AMS, DB, JL, LZ, JC

PP04-2 24976 2.0000 FRI 105 A 3,4,4'-Trichlorocarbanilide Exposure Induces Gut Microbial Dysbiosis in Neonatal Rats 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Endocrine Disruptors Friday, April 1st 11:45:00 AM PP04 7763 11:30:00 AM Endocrine Disrupting Chemicals Poster Preview

Yuet-Kin Leung*, Vinothini Janakiram, Jacek Biesiada, Dan Song, Ady Kendler, Mario Medvedovic and Shuk-Mei Ho
University of Cincinnati College of Medicine, Cincinnati, OH


Bisphenol A (BPA) is a known environmental estrogen and has been found biologically active in many systems even at a dose lower than the reference dose set by US Environmental Protection Agency (EPA). However, its potential impact on estrogen-sensitive endometrium remains largely unknown. In this study, we exposed Sprague Dawley rats with different doses of BPA (0 to 25000ug/kg/day, 5 groups) or two doses of ethinyl estradiol (0.05 and 0.5ug/kg/day EE2 as controls) starting from gestation day 6 until one year. We found rats exposed to 250ug/kg/day BPA (BPA250) and 0.5ug/kg/day EE2 (EE2-05) showed abnormal transition in estrous cycle at postnatal (PND) 90. Continuous exposure to EE2-05, 25ug/kg/day BPA (BPA25) or BPA250 in 1-year-old rats showed prolonged estrous phase, suggesting that BPA25 and BPA250 may act like high dose EE2 (EE2-05) in disrupting normal estrous cycling. To further dissect the underlying molecular mechanism, we selected 1-year-old uterus tissues collected at estrous phase for RNA-sequencing. Using content-specific Bayesian clustering and multi-dimension scaling methods, we observed that the gene signatures from BPA25 and BPA250 groups are clustered away from the control group as well as two EE2 groups. Pairwise comparison against the control group reveals significant differentially expressed genes in different BPA dose groups. Eighty significant genes are found in all BPA groups. Within this BPA subset, 42 genes are overlapped with EE2 signature, which is defined by two EE2 groups. In other words, only 38 genes are BPA-specific but not overlapped with EE2 signature. Only 4 genes show inverted U-shape relationship with BPA doses. Surprisingly, 24 out of 38 genes are sensitive to the lowest dose of BPA (2.5ug/kg/day) and higher BPA concentration exposure results in decreasing fold change. Pathway analysis (IPA) suggested that 38 genes are involved in 4 major gene networks and they are associated with five upstream regulators. Interestingly, majority of these genes are resided in plasma membrane or extracellular space. To determine whether the BPA-specific genes are associated with human endometrial cancer, we queried a publicly available survival dataset of uterine corpus endometrioid carcinoma from The Cancer Genome Atlas (TCGA) with our gene signature. Intriguingly, 74 out of 80 genes can significantly segregate poor vs good prognosis group in this cohort of 332 samples with exceptionally high Hazard Ratio (HR=75, P=2.285x10-5). When we stratified with EE2-overlapped vs non-overlapped genes, both subsets of BPA genes can also significantly predict poor vs good survival using the same cohort (overlapped genes HR=13.2 vs non-overlapped genes HR=13.85). Overall these findings strongly suggest that those genes may actively participate in endometrial cancer progression.


Nothing to Disclose: YKL, VJ, JB, DS, AK, MM, SMH

PP04-3 27700 3.0000 FRI 107 A Bisphenol a-Specific Genes As Predictive Markers for Endometrial Cancer Prognosis 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Endocrine Disruptors Friday, April 1st 11:45:00 AM PP04 7763 11:30:00 AM Endocrine Disrupting Chemicals Poster Preview

Susanna Wegner*1, John Wambaugh2, Caroline Ring1 and David J Dix3
1Oak Ridge Institute for Science and Education, Oak Ridge, TN, 2U.S. Environmental Protection Agency, Research Triangle Park, NC, 3U.S. Environmental Protection Agency, Washington, DC


High-throughput assays and predictive pathway models for endocrine bioactivity provide a method for evaluating thousands of chemicals. As predictive tools are optimized and validated for pathway-based chemical screening by regulatory agencies (e.g. http://www.gpo.gov/fdsys/pkg/FR-2015-06-19/pdf/2015-15182.pdf), they provide an opportunity to predict cumulative bioactivity from exposure to multiple chemicals. Here we explore the application of high-throughput bioactivity and exposure data to predict cumulative estrogen receptor (ER) agonist bioactivity at environmentally relevant levels of human exposure. As a proof of concept we assume a dose additive relationship for ER agonist bioactivity and compute a cumulative ratio of exposure/bioactivity. A model for ER agonist bioactivity from EPA’s ToxCast program integrates results from 16 independent high-throughput in vitro assays to quantify bioactivity in the pathway for over 1800 chemicals. In parallel, an exposure model from EPA’s ExpoCast program uses chemical application and production volume data to predict the geometric mean of human oral exposures across the population. Using toxicokinetic data for in vivo-in vitro extrapolation to relate in vitro bioactivity to in vivo exposures, we compared ER agonist bioactivity predicted in ToxCast to aggregate exposures predicted by ExpoCast for 14 chemicals. Median and lower 95 percent confidence interval ToxCast ER model estimates for different levels of biological effect (10%, 50%, and lowest significant change in bioactivity) were compared to median and upper 95 percent confidence level estimates of population geometric mean exposures. The resulting cumulative exposure/bioactivity ratios reflecting different levels of biological effect and model uncertainty spanned almost 8 orders of magnitude. We used results from the subset of 14 chemicals with toxicokinetic data to predict cumulative bioactivity for all 68 non-pharmaceutical chemicals with ER activity in the ToxCast model. Cumulative exposure/bioactivity ratios predicted by median and 95 percent confidence levels of bioactivity and exposure parameters ranged from several orders of magnitude below one to several orders of magnitude above one, reflecting the large degree of uncertainty currently embedded in both models. Future research priorities are to reduce uncertainty in high-throughput prediction of cumulative bioactivity, incorporate population variability, and account for correlated exposures. Given the relationship between ER bioactivity, toxicity pathways and potential adverse reproductive outcomes, the cumulative exposure/bioactivity ratios from this analysis provide a basis for consideration of cumulative hazard and risk of endocrine disruption.  This abstract does not necessarily represent U.S. EPA policy.


Nothing to Disclose: SW, JW, CR, DJD

PP04-4 27508 4.0000 FRI 111 A High-Throughput Screening Provides an Opportunity to Predict the Potential for Additive Endocrine Bioactivity from Cumulative Chemical Exposures 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Endocrine Disruptors Friday, April 1st 11:45:00 AM PP04 7763 11:30:00 AM Endocrine Disrupting Chemicals Poster Preview

Marina Augusto Silveira*1, Isadora C Furigo1, Jose Donato Jr.2 and Renata Frazao1
1University of Sao Paulo, Sao Paulo, Brazil, 2University of Sao Paulo, Sao Paulo - SP, Brazil


Several studies have proposed that Kiss1 neurons mediate the effects of prolactin on the hypothalamus-pituitary-gonadal axis. In rodents, hyperprolactinemia suppresses Kiss1 mRNA levels and inhibits the ovulatory cycle. However, there is no much information available regarding the acute effects of prolactin on the activity of Kiss1 neurons. The present study was performed to determine whether prolactin affects the membrane electrical properties of the Kiss1 neurons located at the anteroventral periventricular nucleus/anterior periventricular nucleus (AVPV) and arcuate nucleus (ARH). Whole-cell patch-clamp recordings were performed in female mice in diestrus (8-12 weeks). Most of the experiments were performed using brain slices obtained from the Kiss1/hrGFP mouse model. In a subset of experiments, Kiss1 neurons were recorded in brain slices obtained from  Kiss1-Cre/GFP/Stat5fl/fl. Solutions containing ovine prolactin (250 nM) were typically perfused for 5 min. For some experiments, the changes in resting membrane potential (RMP) were monitored in the presence of tetrodotoxin and synaptic blockers, or in the presence of wortmannin, TTX and synaptic blockers. The results demonstrated that prolactin administration to the bath induced a hyperpolarization in 4 out 13 AVPV Kiss1 neurons, reducing the RMP in -7.2 ± 1.1 mV. Two out of 13 AVPV Kiss1 neurons were depolarized by prolactin, with an increase in the RMP of
+5.0 ± 3.0 mV. The remaining recorded AVPV neurons were unaffected by prolactin. The AVPV Kiss1 neurons that were hyperpolarized exhibited a significant decrease in the whole-cell input resistance (P = 0.006), while depolarization induced an increase in the input resistance (P = 0.04). Interestingly, prolactin did not affect the membrane properties of the ARH Kiss neurons (P > 0,05). Pharmacological blockade of voltage-gated sodium channels and synaptic inputs demonstrated that prolactin continued to hyperpolarize AVPV Kiss1 neurons, causing a decrease of -4.6 ± 1.2 mV in the RMP (n= 3 out of 11 cells), while no depolarization was observed. Interestingly, selective Stat5a/b deletion did not prevent the effects of prolactin onto AVPV Kiss1 neurons. Prolactin hyperpolarized 4 out 14, or depolarized 2 out of 14 recorded neurons from Kiss1-Cre/GFP/Stat5fl/fl mice. To determine whether the phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K) signaling pathway mediates the prolactin-induced hyperpolarization of AVPV Kiss1 neurons, experiments were performed in the presence of wortmannin, TTX and synaptic blockers. In this condition,  prolactin effects on AVPV Kiss1 neurons were blocked (n= 12 cells, P = 0.6). Our findings provided evidence about a direct STAT5-independent modulation of AVPV Kiss1 neurons by prolactin. We postulate that prolactin recruits PI3K signaling pathway to induce a hyperpolarization in AVPV Kiss1 neurons and therefore modulate reproduction.


Nothing to Disclose: MAS, ICF, JD Jr., RF

PP09-3 26040 3.0000 FRI 188 A Membrane Electrical Properties of Avpv Kiss1 Neurons Are Modulate By Prolactin 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Reproductive Endocrinology Friday, April 1st 11:45:00 AM PP09 7772 11:30:00 AM Role of Growth Factor Signaling in Fertility and Pregnancy Poster Preview

Zaid Mohsen*, Jacob Johnson, David Garcia Galiano and Carol F Elias
University of Michigan, Ann Arbor, MI


Prokineticin receptor 2 (ProkR2) is one of two cognate GPCRs for two secreted proteins (Prok1 and Prok2) and is predominantly expressed in the mammalian central nervous system.  Loss-of-function mutations of PROKR2 in humans contribute to the development of Kallmann Syndrome due to disruption of GnRH neuronal migration (1). Additionally, ProkR2 has been implicated in the neural modulation of GnRH neurons post-migration. To assess the role of Prok2/ProkR2 signaling in the GnRH system, we have generated a new mouse model that expresses Cre recombinase driven by Prokr2 promoter using the CRISPR-Cas9 technology. Expression of Cre recombinase was visualized using the Cre-LoxP system to express tdTomato as reporter gene (ProkR2-tdTom). High number of tdTomato positive cells was seen in the olfactory bulb and subventricular zone of the lateral ventricle. In the cerebral cortex, tdTomato florescence was found in the piriform cortex, in the gustatory and somatosensory areas. Cells positive for Cre recombinase were also detected in moderate amounts throughout the hippocampus. Moderate to high expression was seen in the amygdala-piriform transition area, lateral septum and several hypothalamic sites including the preoptic area, paraventricular nucleus, suprachiasmatic nucleus, dorsomedial nucleus, and the mammillary nucleus. ProkR2-tdTom cells were also found in circumventricular organs such as the median eminence, subfornical organ, and area postrema. Lastly, ProkR2-tdTom was visualized in the theca and parenchyma cells of the ovaries and the smooth muscle walls of the uterus. Subsequently, the expression of tdTomato was compared to ProkR2 mRNA by in-situ hybridization. We also assessed the co-localization of ProkR2-Cre with estrogen receptor-alpha (ERa). Little-to-no overlap of the cells expressing both receptors was observed.

These data are consistent with known literature and support the validity of our new mouse models in studying the ProkR2 signaling system and physiological functions. A valid animal model that expresses Cre-recombinase in ProkR2 cells will allow for further elucidation of the GnRH system including the differential roles of ProkR2 on the neurodevelopment and migration of GnRH neurons as well as the neuroendocrine effects of ProkR2 signaling in the hypothalamo-pituitary-gonadal axis.


Nothing to Disclose: ZM, JJ, DG, CFE

PP09-4 27051 4.0000 FRI 189 A Characterization of a New Mouse Model Expressing Cre Under ProkR2 Promoter 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Reproductive Endocrinology Friday, April 1st 11:45:00 AM PP09 7772 11:30:00 AM Role of Growth Factor Signaling in Fertility and Pregnancy Poster Preview

Cecilia Martin1, Juhyun Oh2, Elizabeth Wu2, Cristina Luongo1, Monica Dentice3, Warner S Simonides4, Domenico Salvatore3, Amy J Wagers2, P Reed Larsen1 and Ann Marie Zavacki*1
1Brigham and Women's Hospital, Boston, MA, 2Harvard University, Cambridge, MA, 3University of Naples Federico II, Napoli, Italy, 4VU University Medical Center Amsterdam, Amsterdam, Netherlands


Thyroid hormone levels are regulated within specific cell types or tissues independent of serum thyroid hormone concentrations by local alterations in the iodothyronine deiodinase enzymes that activate and inactivate thyroid hormone. In injured skeletal muscle an increase in the thyroxine (T4)-activating type 2 deiodinase (D2) leads to higher T3-levels in satellite cells (muscle stem cells) that promotes differentiation and reduces proliferation (1). We have previously found mice with a targeted deletion of D2 (D2KO) exhibit a significant delay in muscle regeneration and a greater number of satellite cells at 15 days post-injury, and that a primary cell culture model enriched in D2KO-derived satellite cells has increased proliferation (1). Our recent studies using Fluorescence Activated Cell Sorting (FACS) to quantitate the number of satellite cells in D2KO versus WT muscle under basal conditions indicate that a difference in the absolute number of satellite cells was not the cause of the increased number of satellite cells in D2KO mouse muscle after injury. One of the current limitations in the use of transplanted muscle stem cells therapeutically is obtaining sufficient satellite cells. We hypothesized that the increased proliferation of D2KO satellite cells might be beneficial in a transplantation scenario and lead to increased myofiber formation. However, when our findings were translated to a murine satellite cell transplantation model our preliminary results show no difference in engraftment and myofiber formation between D2KO- and WT-derived satellite cells, and no evidence of increased satellite cell proliferation. Further investigation using cultured FACS-purified satellite cells indicated no change in myogenic colony formation or proliferation of singly-plated D2KO satellite cells, indicating that the capacity of D2KO-derived satellite cells to self-renew was not altered. When FACS-isolated D2KO satellite cells were differentiated in vitrothere was no difference in the extent of differentiation between WT and D2KO cells. However, D2KO-derived satellite cells formed myotubes with a significantly increased number of nuclei/myofiber, indicating abnormalities in the fusion process. Using FACS analysis we further identified D2-expression in another component of the muscle stem cell niche, the fibro/adipogenic precursor cells (FAPs). These cells provide pro-myogenic signals to satellite cells after injury, and also are the source of the fibroblasts and adipocytes found in dystrophic muscles when muscle regeneration is impaired. Our data indicates that local thyroid hormone metabolism may play multiple important roles in the muscle stem cell niche.


Nothing to Disclose: CM, JO, EW, CL, MD, WSS, DS, AJW, PRL, AMZ

PP11-1 26647 1.0000 FRI 241 A Novel Roles for Thyroid Hormone Activation in the Skeletal Muscle Stem Cell Niche 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Thyroid/HPT Axis Friday, April 1st 11:45:00 AM PP11 7776 11:30:00 AM Thyroid Hormone Action, Cancer and Clinical Thyroid Poster Preview

Petra Mohacsik*1, Ferenc Erdelyi2, Gabor Szabo2, Monika Toth2, Richard Sinko3, Antonio C Bianco4, Csaba Fekete2 and Balazs Gereben2
1Institute of Experimental Med, Budapest, Hungary, 2Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest, Hungary, 3Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest, 4Rush University Medical Center, Chicago, IL


Thyroid hormones (TH) regulate cellular functions by binding to specific nuclear receptors (TR). These are ligand-dependent transcription factors that interact with cis TH response elements (TRE) and sets of co-activators or co-repressors. TH signaling is directly related to intracellular TH availability, which is regulated by TH transporters and deiodinases. Here we report the creation of the new mouse model Thyroid Hormone Action Indicator Mouse (THAIM) that expresses a luciferase (luc) reporter gene under a 3xTRE controlled thymidine kinase promoter, which allows tissue-specific detection of TH action in vivo. Luc-reporter activity can be measured (i) in tissue sonicates, (ii) by imaging live animals after luciferin injection and (iii) by assessing luc mRNA by RT-qPCR . In general, luc-reporter activity exhibited ubiquitous baseline expression level that was multi-fold stimulated by i.p. administration of T4 or T3 in the majority of tissues including different brain regions, bone, liver, heart, thyroid, intestine, mandibular salivary gland and the brown adipose tissue (BAT). For example, T4 administration (5ug/mouse /day for 3 d) evoked a 64-fold increase of luc activity in the BAT; similar effects were observed in vivo by imaging live animals after 1 ug/g bw T3 injection. The THAIM was also useful in detecting changes in TH action induced by physiological stimuli, e.g. cold stress in BAT both in tissue samples and live. The local activation of type 2 deiodinase (D2)-mediated TH signaling in interscapular BAT (iBAT) of THAIM evoked a 2.5-fold increase in luc reporter mRNA expression as assessed by qPCR after 9 h cold-exposure. This response was abrogated by unilateral sympathetic denervation of the iBAT, which also prevented cold-induction of D2 activity. Notably, luc-reporter mRNA expression was unexpectedly elevated by ~2 fold in the denervated iBAT lobe compared to the intact control lobe at room temperature. This was explained by a ~2.5-fold elevation in TR and MCT8 TH transporter mRNAs  in the denervated iBAT lobe, highlighting the importance of TR level and transmembrane T3 transport in TH signaling. Liver-specific TH action was also studied in THAIM treated with GC24, a TRbeta isoform specific ligand in mammals. Treatment of hypothyroid THAIM with 1.5 nM/g bw GC-24 increased TH action by ~2.5 fold in the liver (a TRbeta-predominant tissue) while TH remained largely unaffected in the TRalpha-predominant heart.  In summary, the novel THAIM model allows in vivo and sample based detection and analytical measurement of tissue-specific TH action in the intact setting of endogenously expressed factors regulating TH action and TH availability. THAIM provides an in vivo mouse platform to study endogenous TH signaling as well as the development of novel TH analogues.


Nothing to Disclose: PM, FE, GS, MT, RS, ACB, CF, BG

PP11-2 26500 2.0000 FRI 239 A A Novel Thyroid Hormone Action Indicator Mouse Model Allows Live Detection and Tissue Specificity 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Thyroid/HPT Axis Friday, April 1st 11:45:00 AM PP11 7776 11:30:00 AM Thyroid Hormone Action, Cancer and Clinical Thyroid Poster Preview

Joanna Klubo-Gwiezdzinska*1, Kelli Gaskins2, Lisa Zhang2, Kirk Ernest Jensen3, Vasyl Vasko4 and Electron Kebebew2
1National Institute of Health, Bethesda, MD, 2National Cancer Institute, NIH, Bethesda, MD, 3Uniformed Services University of the Health Sciences, Bethesda, MD, 4USUHS, Bethesda, MD



It has been reported that treatment with metformin is associated with smaller tumor size, higher complete remission rate and longer progression free survival in diabetic patients with differentiated thyroid cancer (DTC). Furthermore, metformin inhibits DTC cell line growth in vitro via down-regulation of mTOR signaling pathway.  Thus, we hypothesized that metformin will affect disease specific survival by decreasing thyroid cancer growth and progression in a metastatic mouse model of thyroid cancer. 


We used NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ mice that had tail vein injection of 160,000 human DTC cells (FTC-133) transfected with a linearized pGL4.51[luc2/CMV/Neo] vector. When lung engraftment was documented with quantification of luminescence, the mice were randomly assigned into 2 groups:

- control group (C), treated with placebo (water via gastric gavage), n = 11 animals (4 males, 7 females)

- metformin (MF) group - treated with 12.5 mg MF in 250 ul of water via gavage, n=10 mice (4 males, 6 females)

We performed weekly imaging studies with Xenogen IVIS.

The mice were followed until they reached endpoints for euthanasia established by our local Animal Advisory Committee. The disease-specific survival time was calculated from the day of starting the experiment until death.  The lung and liver tissue containing metastatic lesions were removed, fixed in 4% paraformaldehyde in 0.1 M phosphate buffer and then paraffin embedded. Representative H-E stained slides containing the cross section of the lung and liver tissue underwent pathologic evaluation. Pulmonary metastatic tumor burden was expressed as the percentage of tumor divided by percentage of total lung tissue evaluated.


Control mice had more rapid tumor growth as compared with MF treated mice.  Weekly bioluminescence ratios were established by normalizing to baseline: week 1 - C 3.88 vs M 1.78, p=0.1; week 2 - C 9.2 vs M 3.13, p=0.056; week 3 - C 71.4 vs M 22.9, p=0.14; week 4 - C 8080 vs M 167, p=0.1. Disease-specific survival was significantly longer in MF group compared with C group (median 36 days vs 29 days, p=0.002, respectively). Pathology evaluation of H-E stained slides revealed that all mice which died before the endpoint of the experiment had tumor involving 70-90% of normal lung tissue.  The experiment was finished at 36 days when remaining mice from control group met euthanasia criteria. The pathology evaluation of the tissues obtained at day 36 of the experiment revealed that the median tumor burden was 90% of lung volume in C group vs 60% in MF group, p=0.01.


Our study suggests that metformin treatment increases disease specific survival by delaying progression of thyroid cancer metastases in vivo. 


Nothing to Disclose: JK, KG, LZ, KEJ, VV, EK

PP11-3 26861 3.0000 FRI 261 A Metformin Delays Thyroid Cancer Progression and Improves Survival in a Metastatic Mouse Model of Thyroid Cancer 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Thyroid/HPT Axis Friday, April 1st 11:45:00 AM PP11 7776 11:30:00 AM Thyroid Hormone Action, Cancer and Clinical Thyroid Poster Preview

Theodora Pappa*1, Sunee Mamanasiri2, Alexandra M Dumitrescu1, Roy E Weiss3 and Samuel Refetoff1
1The University of Chicago, Chicago, IL, 2Ratchaburi Hospital, Ratchaburi, Thailand, 3University of Miami Miller School of Medicine, Miami Beach, FL


Resistance to thyroid hormone (RTH) is characterized by reduced responsiveness of tissues to thyroid hormones (TH). In 85% of the cases, genetic testing reveals mutations in the thyroid hormone receptor beta (THRB) gene (RTHβ). It was previously shown that exposure of unaffected fetuses to the high TH levels of their affected mothers (carrying a THRB gene mutation) is associated with a higher miscarriage rate, as well as low birth weight and suppressed blood TSH (1). The management of subjects with RTHβ during pregnancy is challenging and there is no prospective data on a group of affected pregnant subjects (2).

Our aim was to prospectively study subjects with RTHβ during pregnancy, prenatally determine the fetus’ genotype (affected or normal) and evaluate whether treatment of the mother with antithyroid drugs has an impact on fetal development and early postnatal course, including thyroid function tests (TFTs) at birth. 

Thirteen RTHβ females were prospectively studied in fourteen pregnancies. Genetic testing of the fetus was performed by Sanger sequencing of fetal samples either from chorionic villi or amniotic fluid. The decision to treat was based on maternal TFTs and the fetus’ genotype. No antithyroid treatment was started in females carrying affected fetuses, whereas in case of unaffected fetus, the target was a free T4 not higher than 20% of the upper limit of normal. Treatment was adjusted by monthly monitoring of maternal TFTs.

The RTHβ females harbored nine different THRB gene mutations: A317T (3 subjects), R320C and M334R (2 subjects each), L450H, E460K, V349M, P453T, R429Q and M310L. Two females had a history of miscarriages and three had older affected children. The genotype distribution was 8 unaffected and 4 affected full term deliveries. Two stillbirths were reported, both involving affected fetuses, one of which due to placental insufficiency. No perinatal and postnatal complications were documented besides one case of fetal distress attributed to pneumonia in an affected infant born to an A317T mother. The mean birth weight (±standard deviation) was 3.181 (±0.33) kg; no significant difference was noted between unaffected and affected infants (3.09±0.39 vs 3.33±0.22 kg). Mean TSH levels at birth were 3.26±1.26 uU/ml and were not different between unaffected and affected infants (4.06±0.69 vs 3.06±1.25 uU/mL).

This study emphasizes the role of prenatal diagnosis in guiding the management of RTHβ subjects in pregnancy. Maintaining T4 levels not higher than 20% of the upper limit of normal in case of unaffected fetuses prevented low birth weight and postnatal TSH suppression, expected based on previous data. Study of additional pregnant subjects with RTΗβ will crystallize our knowledge on maternal-fetal interaction in RTHβ and help optimize clinical care of both the mother and the fetus.


Nothing to Disclose: TP, SM, AMD, REW, SR

PP11-4 26016 4.0000 FRI 246 A Prenatal Diagnosis of Resistance to Thyroid Hormone and Its Clinical Implications 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Thyroid/HPT Axis Friday, April 1st 11:45:00 AM PP11 7776 11:30:00 AM Thyroid Hormone Action, Cancer and Clinical Thyroid Poster Preview

Jennifer DuPont*1, Amy McCurley1, Joseph C McCarthy1, Shawn B Bender2, Wendy Baur1, Michael Hill3 and Iris Z Jaffe1
1Tufts Medical Center, Boston, MA, 2Harry S. Truman Memorial Veterans' Hospital, Columbia, MO, 3University of Missouri, Columbia, MO


Blood pressure (BP) rises with age, making hypertension (HTN) the most prevalent cardiovascular disease risk-factor. We found that mice with SMC-specific deletion of the mineralocorticoid receptor (MR-KO) lack the aging-associated rise in BP. Vascular micro-RNA expression profiling revealed that miR-155 is an aging-associated miR that is modulated by SMC-MR. Ingenuity pathway analysis identified Cav1.2, the pore forming subunit of the L-type calcium channel (LTCC), as a potential target of miR-155. We hypothesize that SMC-MR contributes to BP regulation with aging by regulating vascular L-type calcium channel (LTCC) expression/function via regulation of miR-155. Vascular miR-155 expression declines with age in MR-intact mice (p<0.05 vs. young MR-intact) and is increased in aged MR-KO (p<0.05 vs. MR-intact). Luciferase reporter assays reveal that MR significantly decreased miR-155 promoter activity in a dose dependent manner (p<0.05), suggesting that MR negatively regulates the transcription of miR-155. Overexpression of miR-155 in mouse SMC reduced Cav1.2 expression by 45% (p<0.05 vs. ctrl). Patch clamp studies on freshly dispersed mesenteric resistance vessel (MRV) SMC reveal reduced LTCC current density in SMC from aged MR-KO mice (p<0.05 vs. MR-intact) with no difference in SMC from young mice. Fura-2 photometry studies similarly reveal decreased MRV Ca flux in response to BayK (LTCC agonist) only in aged MR-KO (p<0.05 vs. MR-intact). Vascular contraction to BayK is also blunted in aged MR-KO MRV (p<0.05 vs. MR-intact). RNA expression of Cav1.2 is reduced by 60% in aged MR-KO vs. MR-intact MRV (p<0.05). These data suggest that SMC-MR contributes to HTN through regulating Cav1.2 expression/function, via transcriptional repression of miR-155. These results enhance our basic understanding of BP control with aging, and provide support for innovative therapeutic targets to treat aging-associated HTN.


Nothing to Disclose: JD, AM, JCM, SBB, WB, MH, IZJ

OR02-1 26530 1.0000 A Smooth Muscle Cell Mineralocorticoid Receptors Regulate Vascular L-Type Calcium Channels Via microRNA-155 to Contribute to Aging-Associated Hypertension 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 1:15:00 PM OR02 7661 11:45:00 AM Renin-Angiotensin-Aldosterone - Bench to Bedside Oral

Danielle L Cullinane*, Amanda Elizabeth Garza, Jose R Romero, Gail K. Adler, Luminita H Pojoga and Gordon H Williams
Brigham and Women's Hospital, Harvard Medical School, Boston, MA


Mammalian target of rapamycin (mTOR) has been linked to a variety of pathological outcomes including cardiovascular disease (CVD). Recently, it was reported that mTOR signaling is increased in patients with primary aldosteronism. Aldosterone (ALDO), and its mineralocorticoid receptor (MR) regulate salt/water homeostasis and blood pressure, and excess MR activity contributes to CVD. The epigenetic factors, lysine-specific demethylase 1 (LSD1/KDM1A) and NAD-dependent deacetylase sirtuin-1 (SIRT1) are known to negatively and positively, regulate mTOR signaling, respectively. LSD1 is part of a complex that binds to the mTOR promoter region, and represses gene expression. SIRT1 positively regulates mTORC1 function by inhibiting the acetylation of a downstream target; p70 ribosomal S6 kinase (S6K1). ALDO infusion has been shown to decrease SIRT1 expression in rat kidney. Furthermore, our group has shown that LSD1+/- mice have increased plasma ALDO relative to wildtype mice. Thus, we hypothesize that ALDO infusion will modulate LSD1, SIRT1, and mTOR signaling in cardiac tissue. To test this hypothesis, we studied C57/BL6 male mice maintained on a high salt (HS) diet (1.6% Na+). Mice were randomized to the following treatments for 3 weeks: 1) placebo, 2) ALDO (200µg/kg/day), or 3) ALDO + the mineralocorticoid receptor (MR) antagonist eplerenone (EPL), (100mg/µg/day). Animals were sacrificed and protein expression in cardiac tissue was analyzed by Western Blot. ALDO infusion was demonstrated to be effective by significantly decreased plasma renin activity (PRA) on both ALDO and EPL treated groups. ALDO treatment caused a significant 81% reduction in cardiac mTOR expression that was partially rescued by EPL treatment. The downstream targets of mTORC1 and mTORC2, S6K1 and Protein Kinase B (PKB/AKT) were not significantly modified in any treatment group. In contrast to mTOR expression, LSD1 was significantly increased (80%) in response to ALDO while EPL treatment blocked this effect. These data suggest that increases in LSD1 are mediated by ALDO/MR. Conversely, SIRT1 expression had a decreasing trend in response to ALDO infusion; and was significantly increased (90%) in response to EPL treatment. These data demonstrate that ALDO/MR modulates not only mTOR’s expression, but also the expression of LSD1 and SIRT1, consistent with the role of these epigenetic regulators in mTOR signaling. Thus, our data suggest that the substantial interaction between environmental factors and aldosterone’s adverse CV effects may be mediated by one or both of these epigenetic factors. Finally either these epigenetic regulators or other proteins in the mTOR pathway may prove to be promising new candidates linking ALDO and pathophysiological changes in CVD as well as potential therapeutic targets.


Disclosure: GHW: Ad Hoc Consultant, Daiichi Sankyo, Ad Hoc Consultant, Pfizer Global R&D. Nothing to Disclose: DLC, AEG, JRR, GKA, LHP

OR02-2 26900 2.0000 A Aldosterone's Affect on Epigenetic Regulators and mTOR Signaling in the Heart 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 1:15:00 PM OR02 7661 11:45:00 AM Renin-Angiotensin-Aldosterone - Bench to Bedside Oral

Mohammad Zaki Shukri*1, Richard H Karas2, Luminita H Pojoga3, Iris Z Jaffe2, Gail K. Adler3, Gordon H Williams3 and Jose R Romero3
1Brigham and Women's Hospital, Boston, MA, 2Tufts Medical Center, Boston, MA, 3Brigham and Women's Hospital, Harvard Medical School, Boston, MA


Cardiovascular diseases (CVD) and hypertension are more common in young males when compared to premenopausal females; a difference that disappears following menopause. Sex differences in blood pressure also have been reported in rodent models of hypertension. In addition, our group reported that estradiol-deficient, ovariectomized rats are protected against cardiovascular and renal damage mediated by aldosterone (Aldo) and mineralocorticoid receptor (MR) activation; estradiol replacement restores damage. We tested the hypothesis that sex affects the adrenal and vascular responses to angiotensin II (AngII). We studied 293 female and 447 male human subjects from the Hypertensive Pathotype (HyperPATH) study who were carefully monitored and on controlled high- and low-salt diets for one week. We measured plasma Aldo and systolic blood pressure (BP) before and after in vivo infusion of AngII and calculated ΔAldo and ΔBP. Our results show that ΔAldo was significantly greater in female than in male subjects on a low-salt diet (22.9 ± 13.0 vs 15.8 ± 9.98 ng/dL, respectively; P<0001 adjusted for age, serum cortisol, plasma renin activity, K+, and urinary K+) and on a high-salt diet (9.21 ± 5.85 vs 7.63 ± 5.12 ng/dL; P<0.0001 adjusted). Even when assessed by menopausal status (age >50 yr), the differences persisted. These events were associated with a greater ΔBP in females than males on a low salt diet (17.8 ± 13.7 vs 14.4 ± 11.4 mmHg, respectively; P<0.001 adjusted). These data indicate that females show greater adrenal and vascular responses to infused AngII than do males. To assess potential mechanisms for these gender differences, we studied a hypertensive rodent model characterized by low nitric oxide (NO) and increased AngII to mimic the hypertensive milieu. Male and female wistar rats were maintained on a liberal salt diet and given N{omega}-nitro-L-arginine-methyl-ester (L-NAME) to block NO synthase activity for 14 days with AngII administered on the last 3 days. Urinary and plasma Aldo levels were higher in L-NAME/AngII-treated female rats as compared with L-NAME/AngII-treated male rats (n=6/group; P<0.01). Systolic blood pressures were similar in male and female rats. However, treatment with eplerenone (100 mg/kg), an MR antagonist, led to significant reductions in BP in female but not male rats (n=6/group; P<0.05). In addition, the degree of myocardial damage and proteinuria induced by L-NAME/AngII treatment was greater in female versus male rats (n=8/group, P<0.05). Eplerenone protected female and male rats from L-NAME/AngII induced cardiac and renal damage to a similar extent. These results suggest that under conditions of low NO and high AngII, Aldo-mediated renal and cardiac damage is greater in female rats than in male rats despite similar BP levels. Thus we posit that Aldo/MR activation status may, in part, explain the varying effects of estrogen on CVD in women.


Nothing to Disclose: MZS, RHK, LHP, IZJ, GKA, GHW, JRR

OR02-3 25751 3.0000 A Gender Modulates the Adrenal and Vascular Responses to Angiotensin II 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 1:15:00 PM OR02 7661 11:45:00 AM Renin-Angiotensin-Aldosterone - Bench to Bedside Oral

Hadrien Gaël Boyer1, Julien Wils2, Arnaud Arabo3, Céline Duparc4, Isabelle Boutelet5, Herve Lefebvre*6 and Estelle Louiset7
1Inserm U982, Mont-Saint-Aignan, France, 2INSERM U982, Institute for Biomedical Research and Innovation, Mont Saint Aignan, France, 3Rouen University, Mont Saint Aignan, France, France, 4Normandie University, UNIROUEN, INSERM U982, ROUEN, France, 5Inserm U982, Laboratory of Neuronal and Neuroendocrine Differentiation and Communication, Institute for Research and Innovation in Biomedicine, Rouen University, Mont-Saint-Aignan, France, 6Normandie University, UNIROUEN, INSERM U982, Laboratoire Différenciation et Communication Neuronale et Neuroendocrine, Rouen, France, 7Normandie University, UNIROUEN, INSERM U982, Laboratoire Différenciation et Communication Neuronale et Neuroendocrine, ROUEN, France


We have previously shown that, in the human adrenal gland, aldosterone secretion is stimulated by subcapsular mast cells through local release of serotonin. In addition, mast cell conditioned medium increases CYP11B2 expression in the adrenocortical cell line H295R indicating that mast cells release factors which activate adrenal mineralocorticoid synthesis (1). Interestingly, the density of mast cells is increased in aldosterone-producing adenomas (APA). In the present study, we have investigated the regulation of aldosterone secretion in mast cell-deficient C57BL/6 KitW-sh/W-sh mice in comparison with wild type (WT) C57BL/6 mice. In WT mice, adrenal mast cells located in the subcapsular region of the gland were activated by low sodium diet. KitW-sh/W-sh mice submitted to normal sodium diet had basal plasma aldosterone levels similar to those observed in WT animals but exhibited an increase in renal renin mRNA expression. Moreover, in mast cell-deficient mice, low sodium diet was unexpectedly found to induce an exaggerated aldosterone response which was associated with an increase in adrenal aldosterone synthase expression together with enlargement of zona glomerulosa. The enhancement of aldosterone production appeared to result from an increase in adrenal renin and angiotensin type 1 receptor expression. Severe hyperaldosteronism was secondarily causative for an increase in systolic blood pressure and marked hypokalemia which favored polyuria. Activation of the adrenal renin-angiotensin system seems thus to represent a compensatory mechanism aimed at activating aldosterone production in the absence of mast cells. Globally, these results suggest that mast cells play a significant role in the control of aldosterone secretion in mice. It will therefore be relevant to investigate aldosterone secretion in mouse strains displaying an increase in adrenal mast cell density, which may constitute valuable models for the study of the pathophysiology.

1)  Duparc C et al., J Clin Endocrinol Metab. 2015 100(4):E550-60.


Nothing to Disclose: HGB, JW, AA, CD, IB, HL, EL

OR02-4 26510 4.0000 A Mast Cell Deficiency Causes Dysregulation of Aldosterone Secretion in Mice 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 1:15:00 PM OR02 7661 11:45:00 AM Renin-Angiotensin-Aldosterone - Bench to Bedside Oral

Rene Baudrand*1, Jasmine Torrey2, Gordon H Williams2 and Anand Vaidya2
1Pontificia Universidad Catolica de Chile, Santiago, Chile, 2Brigham and Women's Hospital, Harvard Medical School, Boston, MA


Context: Primary aldosteronism (PA) is the most common cause of secondary hypertension and increases cardiovascular morbidity; therefore, case detection for PA is of public health importance. Broad screening for PA using the aldosterone-to-renin ratio (ARR) is recommended; however, specific dietary considerations and cutoffs for serum aldosterone and plasma renin activity (PRA) when measuring the ARR are not well defined.

Objective: To evaluate the hypotheses that dietary sodium intake variability and insufficiently suppressed PRA can result in inaccurate interpretation of the ARR for PA screening.

Methods: 241 untreated stage I hypertensives with ARR > 20, underwent one week of high sodium diet (HS) such that 24h urine sodium was >200 mmol/d, and subsequently one week of low sodium diet (LS), such that 24h urine sodium was <50 mmol/d. All ARR measurements were taken in the morning after overnight supine rest. Subjects were considered to have a “positive screen” for PA if they had ARR > 20 in addition to PRA ≤ 1.0 ng/mL/h and serum aldosterone ≥ 6 ng/dL. PA was considered to be “confirmed” if positive screens also had a 24h urine aldosterone of ≥12 mcg/d on HS.

Results: 33% (79/241) of the population met criteria for a positive screen for PA; the remaining 67% had a negative screen for PA despite having ARR>20 due to a combination of low aldosterone (< 6 ng/dL) and a highly suppressed PRA. When the 79 subjects with a positive screen for PA underwent LS diet, 56% (44/79) no longer met criteria for a positive screen. This subset of “false negative” screens for PA on LS were characterized as having: higher PRA on LS (2.03 ± 1.9 vs. 0.44 ± 0.3 ng/mL/h; P<0.001) and on HS (0.25 ± 0.13 vs. 0.16 ± 0.1 ng/mL/h; P=0.001), Caucasian race predominance (90 vs. 62%; P=0.002), but no difference in serum aldosterone levels, when compared to subjects whose screen remained positive on LS. Multivariable logistic regression showed that odds for false-negative PA screening on LS were associated with a PRA > 0.3 ng/mL/h on HS (OR=3.7 [1.1–12.5]) and Caucasian race (OR = 8.3 [CI 2.1–32.2]). Most notably, among the 48/79 subjects who had confirmed PA, 52% (25/48) no longer met criteria for a positive screen for PA when placed on LS.

Conclusions: Among individuals with untreated stage I hypertension, we observed that substantial inaccuracies in PA screening can occur if dietary sodium balance and absolute values of PRA and aldosterone are not considered. Our findings suggest that case-detection for PA could be markedly improved by: 1) ensuring a high dietary sodium intake when measuring ARR; and 2) interpreting the ARR in the context of a sufficiently suppressed PRA.


Nothing to Disclose: RB, JT, GHW, AV

OR02-5 24395 5.0000 A The Aldosterone-to-Renin Ratio and False-Negative Case Detection for Primary Aldosteronism 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 1:15:00 PM OR02 7661 11:45:00 AM Renin-Angiotensin-Aldosterone - Bench to Bedside Oral

Nada El Ghorayeb*1, Tania L Mazzuco1, Isabelle Bourdeau2, Eric Thérasse1 and Andre Lacroix3
1Centre de Recherche du Centre hospitalier de l’Université de Montréal, Montreal, QC, Canada, 2Centre hospitalier de l’Université de Montréal (CRCHUM), Montreal, QC, Canada, 3Centre de Recherche du Centre hospitalier de l’Université de Montréal, Montreal, QC, Canada


Background: Adrenal vein sampling (AVS) is required to preoperatively identify lateralized source in Primary Aldosteronism (PA).
Objectives: To evaluate the postoperative outcomes of patients undergoing unilateral adrenalectomy (UA) according to AVS lateralization ratios (LR) of aldosterone to cortisol of dominant side to the opposite side (A/C)DOM/(A/C)OPP ≥2 for basal and/or ≥4 post 250 μg bolus ACTH. To compare pre-op and post-op clinical and biochemical parameters according to various basal contralateral aldosterone (CL) suppression ratios.
Methods: 171 successful simultaneous bilateral AVS in a single referral center were retrospectively studied. Further analyses were performed on the 66/80 patients who underwent UA and had evaluable outcome data. CL suppression was determined by the ratio of basal aldosterone of opposite side/periphery AOPP/AP and compared to the ratio of (A/C)OPP/(A/C)P. Clinical improvement (CI) of PA was defined as achieving at least 2 criteria: BP <140/90, decrease in number of antihypertensive medications by ≥50%, normokalemia without need for supplements; clinical cure (CC) as BP <140/90 and normokalemia without any medications and biochemical cure (BC) by normalization of aldosterone/renin ratio.
Results: Using a post ACTH LR≥4 criteria, 33.8% displayed bilateral disease, 28.7% lateralized to the right and 37.6% to left. A discordance of lateralization between basal (LR≥2) and post-ACTH (LR≥4) values was observed in 28% of cases. Of the 66 patients who underwent UA, 26% achieved CC, 61% BC, 17% both, and 58% CI with significant change in post-op BP, K, PAC, renin and number of medications (p<0.001). In the subgroup (85% of UA patients) lateralizing basally (LR≥2) and post ACTH (LR≥4): 27% achieved CC, 55% BC, 16% both, and 55% CI. Among the 8 patients who lateralized only basally:  7 achieved BC, 6 CI, and 2 CC. The 2 patients lateralizing only post ACTH achieved BC and CI but not CC. 77% of UA patients displayed CL suppression using (A/C)OPP/(A/C)P <1.5. vs 30% only when using the AOPP/AP at the same cut-off. Patients in the group with CL suppression had more severe form of PA ie higher BP and PAC with lower pre-op K compared to non-suppressed group (p=0.02).  Absence of CL suppression was associated with lower rate of response to UA in terms of clinical and biochemical parameters with difference in CI (35% vs 67% p=0.01), CC (55% vs 13% p=0.0003) and overall cure (35% vs 9% p=0.0003) using AOPP/AP. In contrast, no difference noted for CC or BC when using (A/C)OPP/(A/C)P.
Conclusions: Although ACTH stimulation is useful to improve selectivity of AVS, it can modify interpretation of lateralization in 28% of cases; therefore basal ratios are as important as post ACTH ratios to set an indication of UA. CL suppression is a valuable tool in AVS interpretation and helps predict post-op outcomes and potential cure from PA. AOPP/AP provided superior results compared to (A/C)OPP/(A/C)P


Nothing to Disclose: NE, TLM, IB, ET, AL

OR02-6 24825 6.0000 A Importance of Basal and Post ACTH Aldosterone Levels and Contralateral Aldosterone Suppression during Bilateral Simultaneous Adrenal Venous Sampling in Primary Aldosteronism 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 1:15:00 PM OR02 7661 11:45:00 AM Renin-Angiotensin-Aldosterone - Bench to Bedside Oral

Jean-David Gothié*1, Marine Perret-Jeanneret1, Sylvie Remaud1 and Barbara Demeneix2
1Museum national d'histoire naturelle, France, 2Muséum National d'Histoire Naturelle, Paris CEDEX 05, France


Thyroid hormones (THs) are implicated in differentiation and maturation of both neurons (1) and oligodendrocytes (2). In particular, THs and Thyroid hormone receptor α (TRα) also regulate neural stem cell (NSC) proliferation and commitment towards a neuronal phenotype in the adult murine subventricular zone (SVZ), one of the major neurogenic niches in the mammalian brain (1). It is well established that THs can also regulate metabolic activities in several tissues (3). Stem cells display metabolic features of aerobic glycosylation rather than oxidative phosphorylation (OXPHOS), in contrast to committed, differentiated cells (4). Growing evidence shows that changes in metabolic status influence stem cell pluripotency and differentiation. Given the major implications of THs in both NSC fate choice and mitochondrial metabolism, we are testing the hypothesis that THs modulate the transition from glycolytic to OXPHOS metabolism and thus could be critical for NSCs cell fate decision.

First, we analysed mitochondrial function in vivo in the different cell types of the adult mouse SVZ. Following stereotaxic injection of a mitochondrial membrane potential dye, JC-1, into the lateral ventricle of the brain, we observed greater mitochondrial activity in DCX+ neuroblasts than in EGFR+ proliferative progenitor cells and NG2+ oligodendrocyte precursors. Furthermore, preliminary data suggest that a short-term hypothyroidism also affects mitochondrial activity in the SVZ. Second, we studied the expression pattern of the activated form of the DRP1 protein (pDRP1S616). It has been shown that THs can induce the translocation of DRP1 to the mitochondria (5), where it can mediate mitochondrial fission, and in turn impact mitochondrial respiration. We showed that pDRP1S616 is preferentially present in the cytosol of cells differentiating toward a neuronal fate in the adult SVZ.

Taken together, the results show that THs direct NSCs differentiation toward a neuronal rather than an oligodendroglial phenotype in association with metabolic modulation. We hypothesize that it is through their impact on mitochondrial activity that THs govern NSCs commitment. Future work addressing the interplay between THs and mitochondrial metabolism underlying this cell fate decision will involve studying the impact of modulating DRP1 in vivo and in vitro in different thyroid contexts. This work should provide new insights on molecular and cellular mechanisms governing adult NSCs fate in control conditions and in response to changing energy metabolism.


Nothing to Disclose: JDG, MP, SR, BD

OR05-1 25789 1.0000 A Thyroid Hormones and Cell Mitochondrial Metabolism Cooperate in Determining Adult Neural Stem Cell Fate 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 1:15:00 PM OR05 7664 11:45:00 AM Neuroendocrinology Oral

Hanne Mette Hoffmann*1, Jason D Meadows1, Crystal Trang1, Brittainy Hereford1, Kapil Bharti2, Michael R Gorman1 and Pamela L Mellon1
1Center for Circadian Biology, University of California, San Diego, La Jolla, CA, 2NIH, Bethesda, MD


Modern life has enabled humans the opportunity to travel rapidly across time zones, to manipulate the light environment and work late into the night – all of which can disrupt sleep and circadian rhythms.  Today more than 20% of the population suffers from disrupted sleep patterns, which affect circadian rhythms and impair fertility. Female fertility is particularly sensitive to compromised circadian rhythms causing dysregulation of the hypothalamic-pituitary-ovarian axis (HPO). Formation of the suprachiasmatic nucleus (SCN) is a fundamental step in the establishment of circadian rhythms, and is required for fertility. The SCN orchestrates the HPO axis by coordinating gonadotropin-releasing hormone (GnRH) neuron activity with peripheral tissues and light cycles. Mature GnRH neurons release GnRH in a pulsatile fashion maintaining appropriate levels of sex steroids across the estrous cycle and the preovulatory LH surge. We here identify a novel homeoprotein required for SCN development and function, Ventral Anterior Homeobox 1 (VAX1). During embryogenesis, VAX1 is necessary for the establishment of normal SCN development and expression of the SCN transcription factor, SIX homeobox 3 (SIX3), as well as two core SCN peptides, arginine vasopressin and vasoactive intestinal peptide, which demarcate the SCN shell and core regions respectively. Interestingly, in the adult brain VAX1 is almost exclusively expressed in the SCN. Since the Vax1 knock-out mouse is perinatal lethal, we generated Vax1flox mice and crossed them with synapsincre mice, thus deleting VAX1 in mature neurons in the brain. Vax1flox:synapsincre mice had impaired wheel running activity in constant darkness, which correlated with abnormal expression of SCN peptides, indicating weak SCN output. Indeed, Vax1flox:synapsincre mice had prolonged estrous cycles and decreased fertility. Surprisingly, synapsincre deletion of SIX3, a downstream target of VAX1 during development, led to behavioral arrhythmicity in constant darkness, irregular estrous cycles, increased ovulation and almost complete infertility along with abnormal LH levels in constant darkness and a lack of estrogen-induced LH surge in ovariectomized females. This suggest that SIX3 and VAX1 play distinct roles in the SCN in adulthood. VAX1 modestly regulates SCN output and fertility, whereas SIX3 is a strong regulator of SCN function and female fertility. This study identified VAX1 as critical for SCN development as well as normal SCN function in adulthood. VAX1 is required for normal wheel running activity, SCN peptide expression and estrous cyclicity. More strikingly, we found that deletion of SIX3 in mature neurons completely impaired SCN function and led to almost complete infertility due to abnormal ovarian function.


Nothing to Disclose: HMH, JDM, CT, BH, KB, MRG, PLM

OR05-2 24122 2.0000 A Deletion of SIX3 or VAX1 in the SCN Impairs Circadian Rhythms and Fertility 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 1:15:00 PM OR05 7664 11:45:00 AM Neuroendocrinology Oral

Leonardo Domingues Araújo*1, Silvia Ruiz Roa2, Ana Carolina Bueno1, Fernanda B. Coeli-Lacchini1, Ernane Torres Uchoa1, Ayrton C. Moreira1, Jose Antunes-Rodrigues1, Lucila Elias1 and Paula C. L. Elias1
1Ribeirao Preto Medical School - University of Sao Paulo, Ribeirao Preto, Brazil, 2Ribeirao Preto Medical School, University of Sao Paulo, Ribeirao Preto, Brazil


Introduction: Food restriction can change the expression of clock genes. Objective: To evaluate Clock, Bmal1, Per1, Per2, Per3, Cry1, Cry2, Rev-erbα and Rorα expression in hypothalamic nuclei of Wistar rats on feeding restriction schedules. Material and Methods: Control group (CG): food ad libitum; Food Restriction (FR): food available from Zeitgeber time (ZT) 12-14 (6-8PM); and Food Shift (FS): food available from ZT3-5 (9-11AM) kept for 21 days with light/dark cycle (lights on 6AM–ZT0). Rats were decapitated at ZT3, ZT11, and ZT17. Corticosterone was measured (RIA). Suprachiasmatic (SCN), Paraventricular (PVN) and Arcuate (ARC) hypothalamic nuclei were microdissected (punch). Gene expression was determined by qPCR (2-ΔΔCT). Results: CG showed greater weight (g) and daily food intake (g) compared to FR and FS groups (385.4±55.4 vs 245.5±32.0 vs 227.9±40.9) and (31.2±3.9 vs 14.1±1.8 vs 13.3±1.6), respectively (P<0.0001). CG presented higher corticosterone levels (mg/dl) at ZT11 (14.1±8.0) and ZT17 (12.9±6.2) compared to ZT3 (1.0±0.6)(P=0.0009). FR showed higher corticosterone levels at ZT11 (20.7±7.6) compared to ZT3 (3.6±2.6) and ZT17 (3.5±2.6)(P<0.0001). FS showed an inverted corticosterone pattern with higher levels at ZT3 (22.7±6.2) compared to ZT11 (10.6±5.7) and ZT17 (5.6±3.3) (P=0.0002). In the SCN of CG, Per2 expression was higher at ZT11 while in FS and FR, its expression was also higher at ZT3 (P=0.0005). No differential Clock, Per1, and Per3 expressions were observed in CG, but higher expression was seen at ZT3 and ZT3 in FS and FR (P=0.002). Compared to CG and FR groups, FS presented higher Bmal1 expression at ZT11, and higher Cry1, Cry2 and Rorα expression at ZT3 and ZT11 (P=0.008). PVN of CG showed higher Per1 expression at ZT11 and ZT17 (P=0.002), while Per2, Per3, Cry2, and Rorα showed higher expression at ZT17 (P=0.02). In FR, the nocturnal CG pattern of Per2, Per3, and Rorα expression was maintained (P=0.04) while Per1 and Cry2 was lost. In FS, there was no differential expression of all studied genes among ZT3, ZT11, and ZT17, but Rev-erbα had higher expression at ZT17 (P=0.03), demonstrating lost of the CG pattern in FS. In ARC of CG, Cry1 expression was higher at ZT3; Per1 was higher at ZT11 and ZT17; while Clock, Per2, Per3, Cry2, Rev-erbα, and Rorα were higher at ZT17 (P=0.007). FR and FS groups showed similar CG pattern of Clock, Per3, Cry2, and Rorα expression (P=0.002). FR also had higher Per1 expression at ZT11 (P=0.02), while the pattern of Per2, Cry1, and Rev-erbα was lost. Differently to CG, which had higher expression of Per2 at ZT17 and Cry1 at ZT3, FS group lost these patterns with no difference at ZT3, ZT11, and ZT17. Higher Per1 expression was observed at ZT3, in a modified pattern compared to CG and FR groups. Conclusion: Restricted feeding schedules modulate clock genes in the SCN, as well as in PVN and ARC nuclei, these latter involved in energy homeostasis and food motivation.


Nothing to Disclose: LDA, SRR, ACB, FBC, ETU, ACM, JA, LE, PCLE

OR05-3 25940 3.0000 A Influence of Restricted Feeding Schedules in the Expression of Clock Genes in the Hypothalamus 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 1:15:00 PM OR05 7664 11:45:00 AM Neuroendocrinology Oral

Dakota Clinton Jacobs*1, Rebecca Veitch1 and Patrick Everett Chappell2
1Oregon State University, Corvallis, OR, 2Oregon State University College of Veterinary Medicine, Corvallis, OR


Mammalian female reproduction requires preovulatory surges of gonadotropin-releasing hormone (GnRH) from neurons in the hypothalamic preoptic area (POA), initiated by elevated ovarian estradiol (E2). Rising E2 activates a subset of sexually dimorphic kisspeptin (Kiss1) neurons in the female, located in the anteroventral periventricular nuclei (AVPV). Conversely, E2 negative feedback on GnRH secretion is mediated by a neuroanatomically separate population of Kiss1 neurons in the arcuate nuclei. Kisspeptin stimulates GnRH expression and secretion in vivo, and the development of this system is critical for the initiation of puberty. To elucidate how phenotypically similar Kiss1 neuronal populations react differentially to E2 exposure, we have generated two immortalized kiss1-expressing and -releasing cell lines from kiss1-GFP post-pubertal female mice. In addition to expressing kiss1, these neuronal cell lines are GFAP-negative, and both express pro-dynorphin (pdyn) and NKB (tac2), with higher expression observed in arcuate-derived KTaR-1 cells. These cell models recapitulate in vivo differential responsiveness to 17β-E2, with KTaV-3 (AVPV-derived) demonstrating ~5-fold increases in kiss1 expression under higher E2 doses (5pM – 50pM), while kiss1 expression in KTaR-1 cells is suppressed up to 80% under lower E2 concentrations (2pM – 10pM). Additionally, both cell lines secreted kisspeptin into the media, as evaluated by ELISA. 50pM E2 stimulated KISS1 secretion from KTaV-3 cells, and inhibited secretion from KTaR-1 cells after 8 hours. Further, we have found that baseline expression of estrogen receptors α (ERα/esr1) and β (ERβ/esr2) is significantly different between KTaV-3 and KTaR-1 cells, with KTaV-3 exhibiting 5-fold higher basal expression of esr2 and KTaR-1 showing equivalently higher basal expression of esr1. This relative expression difference between esr1 and esr2 in KTaV-3 and KTaR-1 cells was observed to shift after 25pM E2 exposure, with significant elevations in esr1 expression observed in KTaV-3 cells compared to KTaR-1 cells after 4 hours, accompanied by concomitant decreases in esr2 expression. We have begun exploring temporal patterns of kiss1 and core clock gene expression in these lines in response to E2, and found distinct antiphasic patterns of bmal1 and per2 in KTaV-3 cells irrespective of E2 exposure. Treatment of KTaV-3 cells with 25pM E2, however, elicited distinct patterns of kiss1 expression over time in contrast to vehicle treatment, suggesting a differential coupling of intracellular oscillators to kiss1 transcriptional activity in the presence of E2. Ongoing delineation of responsiveness to E2 in these lines could reveal novel molecular mechanisms underlying observed differential expression patterns demonstrated in vivo between these distinct Kiss1 neuronal populations.


Nothing to Disclose: DCJ, RV, PEC

OR05-4 27118 4.0000 A Evaluation of Immortalized Avpv- and Arcuate-Specific Neuronal Kisspeptin Cell Lines Reveal Potential Mechanisms of Differential Responsiveness to Estrogen 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 1:15:00 PM OR05 7664 11:45:00 AM Neuroendocrinology Oral

Cari Nicholas*1, Andrew Wolfe2, Streamson C Chua3 and Genevieve S. Neal-Perry4
1Albert Einstein College of Medicine, Bronx, NY, 2Johns Hopkins University School of Medicine, Baltimore, MD, 3Albert Einstein Coll of Med, New York, NY, 4University of Washington, Seattle, WA


We previously reported that maternal (in utero and preweaning) vitamin D (VitD) deficiency programs estrous cycle dysfunction that is characterized by extended periods of diestrus and attenuated preovulatory LH surges in female offspring. The mechanisms by which maternal VitD deficiency programs female reproductive dysfunction is minimally understood. The integration of metabolic cues by the POMC and NPY neurons, a neuronal network that controls fertility, is critical for reproduction in mammals. Of note, the online motif finder MEME Suite software indicates that both Pomc (AGATTA TT GGGTTTA) and Npy (AGCTTG TG AAGTTCA) have VDRE consensuses located in their upstream regulatory regions. Additionally, the POMC post translational derivative, αMSH, activates GnRH neurons and induces LH release. In contrast, NPY predominantly inhibits GnRH stimulated LH release. We hypothesize that maternal VitD deficiency programs reproductive axis dysfunction in female offspring, in part, through adverse effects on the POMC and NPY neurotransmitter system.

Methods: To determine the effect of maternal diet on hypothalamic gene expression (n = 5) of Pomc, Mc4r (melanocortin 4 receptor; αMSH receptor), and Npy in female offspring exposed to maternal VitD deficiency, we used qRTPCR. To determine if exposure to maternal VitD deficiency affected hypothalamic responsiveness to excitatory peptide, αMSH, we administered MTII (αMSH analog) via an intraperitoneal injection to female offspring exposed to control (n=7) or maternal VitD deficient diets (n=5) and collected serial blood samples to measure serum LH levels.

 Results: Compared to controls, female offspring exposed to maternal VitD deficiency exhibited attenuated LH release in response to MTII (13.69 vs 7.03 ng/ml; p< 0.005). Reduced responsiveness to MTII was not associated with gross changes in hypothalamic, Mc4r, mRNA expression. Female offspring exposed to maternal VitD deficiency had a 0.5-fold reduction in hypothalamic Pomc (p< 0.05) and a 3-fold increase in Npy mRNA expression (p< 0.05).

Conclusion: Maternal VitD deficiency disrupts expression of hypothalamic Pomc and Npy mRNA and responsiveness to MTII. These findings suggest that in utero VitD signaling may act as a metabolic cue that is important for the programming of the reproductive neuroendocrine axis of adult offspring.  Moreover, these data suggest maternal VitD deficiency may program female reproductive axis dysfunction through adverse effects on the balance of neuropeptides that transmit information about energy homeostasis.


Nothing to Disclose: CN, AW, SCC, GSN

OR05-5 27331 5.0000 A Maternal Vitamin D Deficiency Disrupts GnRH Neuron Responsiveness to Alpha MSH in Affected Adult Female Offspring 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 1:15:00 PM OR05 7664 11:45:00 AM Neuroendocrinology Oral

Rim Hassouna*, Moneek Madra and Lori Michelle Zeltser
Columbia University, New York, NY


Abstract: Anorexia nervosa (AN) is a complex disease characterized by compulsive restriction of food intake. As current medications do not impact eating behaviors that drive AN’s high mortality rate, there is an urgent need to identify new therapeutic targets. While model systems are usually helpful in these circumstances, the critical involvement of psychosocial risk factors has been an obstacle to developing reliable animal models of AN that approximate the human disease. We developed a novel mouse model study the onset of AN-like behavior by combining factors that are associated with increased risk of AN – adolescent females, genetic predisposition to anxiety imposed by the Brain-derived growth factor (BDNF)-Val66Met gene variant, social stress and caloric restriction. Approximately 40% of the mice with all of these risk factors exhibit severe self-imposed dietary restriction (i.e. food intake of less than 0.5g within 24h as compared to 3-4g for controls).

By comparing age- and exposure-matched mice that exhibited anorexia-like behavior from those that did not, we are beginning to identify neuroanatomical and molecular correlates of subsequent risk of anorexic behavior. We found that levels of arginine vasopressin (AVP) in the serum were more than 2-fold higher in mice that previously exhibited anorexic behavior, similar to observations in AN patients (1). Moreover, we found that exposure of Val66Met carriers to social isolation stress was associated with increased AVP levels in the serum before the onset of aphagic behavior. We then explored whether increased risk of anorexic behavior was also correlated with changes in other components of the BDNF and AVP signaling pathways in the brain. We identified marked increases in the expression of genes encoding the AVP receptor, Avpr1a, and the BDNF p75NTR receptor (Ngfr) in the amygdala, while expression in the hypothalamus, prefrontal cortex, hippocampus and pituitary were not affected. The idea that AVPR1A signaling is predictive of and contributes to severe dietary restriction is supported by pharmacological experiments in animals (2, 3) and observations that a variant in the AVPR1A gene that is associated with increased risk of severe dietary restriction in humans is also associated with increased amygdala reactivity (4). This study raises the possibility that developmental impacts on the AVP system could be exploited to develop novel therapeutic compounds to prevent and/or treat AN.


Nothing to Disclose: RH, MM, LMZ

OR05-6 27634 6.0000 A Role of the Arginine-Vasopressin Tone in a Mouse Model of Susceptibility to Anorexia-Nervosa 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 1:15:00 PM OR05 7664 11:45:00 AM Neuroendocrinology Oral

B Langdahl*1, C Libanati2, D B Crittenden3, M A Bolognese4, J P Brown5, N S Daizadeh3, E Dokoupilova6, K Engelke7, J S Finkelstein8, H K Genant9, S Goemaere10, L Hyldstrup11, E Jodar-Gimeno12, T M Keaveny13, David L Kendler14, P Lakatos15, J Maddox3, J Malouf16, F Massari17, J F Molina18, M R Ulla19 and A Grauer3
1Aarhus University Hospital, Aarhus, Denmark, 2UCB Pharma, Brussels, Belgium, 3Amgen Inc., Thousand Oaks, CA, 4Bethesda Health Research Center, Bethesda, MD, 5Laval University and CHU de Québec (CHUL) Research Centre, Québec City, QC, Canada, 6Medical Plus, Uherske Hradiste, Czech Republic, 7BioClinica, Inc., Hamburg, Germany, 8Department of Medicine, Massachusetts General Hospital, Boston, MA, 9Department of Radiology, University of California, San Francisco, CA, 10Ghent University Hospital, Gent, Belgium, 11Hvidovre University Hospital, Hvidovre, Denmark, 12Servicio de Endocrinología, Hospital Universitario Quirón, Madrid, Spain, 13University of California at Berkeley, Berkeley, CA, 14University of British Columbia, Vancouver, BC, Canada, 15Department of Medicine, Semmelweis University, Budapest, Hungary, 16Universitat Autònoma de Barcelona, Barcelona, Spain, 17Instituto de Investigaciones Metabólicas, Buenos Aires, Argentina, 18Reumalab Centro Integral de Reumatologia, Medellin, Colombia, 19Instituto Latinoamericano de Investigaciones Médicas, Cordoba, Argentina


Purpose: STRUCTURE was a phase 3, open-label study evaluating the effect of romosozumab or teriparatide (TPTD) for 12 months in women with postmenopausal osteoporosis (PMO) transitioning from bisphosphonate therapy (NCT01796301).

Methods: This multi-center, active-controlled study enrolled women with PMO who had taken an oral bisphosphonate for ≥ 3 years prior to screening and alendronate (70 mg weekly or equivalent) in the year prior to screening; had a bone mineral density (BMD) T-score ≤ –2.5 at the total hip (TH), lumbar spine (LS), or femoral neck (FN); and had a history of nonvertebral fracture after age 50 or vertebral fracture. Subjects received daily calcium and vitamin D and were randomized to receive subcutaneous romosozumab 210 mg once monthly or TPTD 20 μg once daily. The primary endpoint was percent change from baseline in BMD by DXA at the TH through month 12 (the average at months 6 and 12). Secondary endpoints included percent change from baseline at months 6 and 12 in BMD by DXA at the TH, LS, and FN; hip integral and cortical BMD by quantitative computed tomography (QCT); and estimated hip strength by finite element analysis. Imaging assessments were done blinded to treatment.

Results: A total of 436 women were randomized to receive romosozumab (N = 218) or TPTD (N = 218) with a mean age of 72 years. Baseline mean TH, LS, and FN T-scores were –2.2, –2.9, and –2.5, respectively. Through 12 months, the mean (95% CI) percent change from baseline in TH BMD by DXA was 2.6% (2.2, 3.0) with romosozumab and –0.6% (–1.0, –0.2) with TPTD (p < 0.0001 between groups). TH BMD changes at months 6 and 12 were significantly larger with romosozumab than with TPTD (p < 0.0001): month 6, 2.3% (1.9, 2.7) vs –0.8% (–1.2, –0.4); month 12, 2.9% (2.5, 3.4) vs –0.5% (–0.9, 0), respectively. Romosozumab also resulted in significantly larger BMD gains at the LS at months 6 and 12 vs TPTD (p < 0.0001): month 6, 7.2% (6.6, 7.8) vs 3.5% (2.9, 4.0); month 12, 9.8% (9.0, 10.5) vs 5.4% (4.7, 6.1), respectively. QCT assessments of the hip demonstrated significantly greater gains in integral and cortical BMD with romosozumab vs TPTD at months 6 and 12 (p < 0.0001). Estimated hip strength gains were also significantly larger with romosozumab at both time points (p < 0.0001) and declined from baseline in TPTD-treated subjects at month 6. The subject incidences of treatment emergent adverse events and adverse events of interest were generally balanced between treatment groups.

Conclusions: In subjects transitioning from bisphosphonate therapy, romosozumab was well-tolerated and was associated with significant BMD gains at both the hip and spine compared with TPTD. BMD gains in the cortical compartment contributed to the greater treatment effect of romosozumab at the hip. Estimated hip strength improved with romosozumab over 12 months but decreased early with TPTD. A global phase 3 program evaluating romosozumab for the treatment of PMO is ongoing.


Disclosure: BL: Speaker, Amgen, Eli Lilly, Merck, Investigational drug, Eli Lilly, Orkla, Advisor, Amgen, Eli Lilly, Merck, UCB Pharma. CL: Employee, UCB Pharma, Employee, UCB Pharma, Employee, UCB Pharma. DBC: Employee, Amgen, Employee, Amgen, Employee, Amgen. MAB: Study Investigator, Amgen, Lilly, Pfizer, Sanofi, Speaker Bureau Member, Amgen. JPB: Speaker, Amgen, Eli Lilly, Study Investigator, Amgen, Eli Lilly, Advisory Group Member, Amgen, Eli Lilly, Merck. NSD: Employee, Amgen, Employee, Amgen, Employee, Amgen. ED: Principal Investigator, Amgen. KE: Employee, BioClinica. HKG: Consultant, Amgen, Janssen, Lilly, Merck, Roche, Synarc. SG: Speaker, Amgen, Principal Investigator, Amgen, MSD, Novartis, Principal Investigator, MSD. LH: Teacher, Amgen, Denmark; Eli-Lilly, Denmark. EJ: Study Investigator, Amgen, Lilly, Speaker, Amgen, Lilly, MSD, Ad Hoc Consultant, Amgen, MSD. TMK: Owner, O.N. Diagnostics, Consultant, Agnovos, Amgen, O.N. Diagnostics. DLK: Medical Lecture, Amgen, Eli Lilly, GSK, Principal Investigator, Astalis, AstraZeneca, Contract Research, Amgen, Eli Lilly, Advisory Board, Amgen, Eli Lilly, Pfizer, Safety monitoring board, Merck & Co.. PL: Speaker, Amgen, Lilly, Servier. JM: Employee, Amgen, Employee, Amgen, Employee, Amgen. JM: Speaker, Gruenenthal, Principal Investigator, Amgen, Lilly España. JFM: Investigator, Amgen. AG: Employee, Amgen, Employee, Amgen, Employee, Amgen. Nothing to Disclose: JSF, FM, MRU

OR01-1 23904 1.0000 A Superior Gains in Bone Mineral Density and Estimated Strength at the Hip for Romosozumab Compared with Teriparatide in Women with Postmenopausal Osteoporosis Transitioning from Bisphosphonate Therapy: Results of the Phase 3 Open-Label Structure Study 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 1:15:00 PM OR01 7684 11:45:00 AM Osteoporosis: What You Had, What You Lost, and What You Gain Oral

Felicia Cosman*1, Gary Hattersley2, Paul D Miller3, Ming-yi Hu4, Luis Augusto Russo5, Bente Juel Riis6, Gregory C Williams2 and Lorraine A Fitzpatrick7
1Helen Hayes Hospital, West Haverstraw, NY, 2Radius Health, Inc, Waltham, MA, 3Colorado Center for Bone Research, Lakewood, CO, 4Radius Health Inc, Waltham, MA, 5CCBR Rio De Janeiro, Rio de Janiero, Brazil, 6Nordic Biosciences, Copenhagen, Denmark, 7Radius Health, Inc, Wayne, PA


The investigational drug abaloparatide is an osteoanabolic analogue of PTHrP (1-34) that is being studied for potential use in the treatment of postmenopausal osteoporosis. ACTIVE (Abaloparatide Comparator Trial In Vertebral Endpoints) was a trial of 2463 postmenopausal women with osteoporosis (aged 49-86 years; mean=69 years old) who were randomized to double-blinded abaloparatide-SC 80 µg or placebo, or open-label teriparatide 20 µg SC for 18 months. At 18 months, the data show that abaloparatide increased BMD from baseline at the lumbar spine 9.2%, total hip 3.4% and femoral neck 2.9% (all p<0.0001 vs placebo).  Abaloparatide reduced morphometric vertebral fractures 86% (p<0.0001), nonvertebral fractures 43% (p=0.0489) and major osteoporotic fractures 70% (p=0.0004) compared to placebo and reduced major osteoporotic fractures compared to teriparatide by 55% (p=0.0309).  Prespecified subgroup analyses were performed to evaluate if fracture risk reduction was consistent across different levels of baseline risk.  Risk factor subgroups were defined categorically for BMD T-score of the lumbar spine, total hip and femoral neck (≤-2.5 vs >-2.5 and ≤-3.0 vs >-3.0), fracture history (yes vs no), prevalent vertebral fracture (yes vs no) and age (<65 vs 65 to <75 vs ≥75 years old) at baseline.  Results of forest plots show consistent fracture reduction in the abaloparatide arm for new morphometric vertebral or nonvertebral fractures without any interactions caused by baseline risk factors. Furthermore, there were no interactions between any of the baseline risk factors and magnitude of BMD accrual by abaloparatide.  In conclusion, these data suggest that abaloparatide may have potential to provide protection against fractures consistently across a wide variety of ages and baseline risks, including those with and without prior fractures, as well as utility for a broad group of patients with osteoporosis.


Disclosure: FC: Consultant, Amgen, Consultant, Eli Lilly & Company, Consultant, Merck & Co., Consultant, Radius Health, Inc. GH: Chief Scientific Officer, Radius Health, Inc. MYH: Coinvestigator, Radius Health, Inc. LAR: Clinical Researcher, Center for Clinical and Basic Research. BJR: Consultant, Nordic Biosciences. GCW: Chief Operating Officer, Radius Health, Inc. LAF: Chief Medical Officer, Radius Health, Inc. Nothing to Disclose: PDM

OR01-2 27411 2.0000 A Abaloparatide Significantly Reduces Vertebral and Nonvertebral Fractures and Increases BMD Regardless of Baseline Risk 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 1:15:00 PM OR01 7684 11:45:00 AM Osteoporosis: What You Had, What You Lost, and What You Gain Oral

Arun S Karlamangla*1, Albert Shieh1, Sherri-Ann M Burnett-Bowie2, Elaine W. Yu2, Gail A Greendale1, Patrick M. Sluss2, Deborah Martin3 and Joel S Finkelstein2
1University of California, Los Angeles, CA, 2Massachusetts General Hospital, Boston, MA, 3University of Pittsburgh, Pittsburgh, PA


The menopause transition (MT) in women is a period of bone loss, with the most rapid declines occurring in a 3-year period bracketing the final menstrual period (FMP). This period of rapid bone loss has been called the trans-menopause, and the rate of BMD decline over this period varies substantially between women (1).  Circulating levels of Anti-Mullerian Hormone (AMH) made by ovarian granulosa cells also decline as women progress through the MT (2). We hypothesized that serum levels of AMH in women early in the MT will predict the rate of bone loss over the trans-menopause. We tested this hypothesis using data from The Study of Women’s Health Across the Nation, a 7-site, multi-ethnic study of the MT. At baseline, participants had to be 42 to 52 years old, pre- or early peri-menopausal, have an intact uterus with 1 or 2 ovaries, and not be taking exogenous sex steroid hormones. Enrollment began in 1996 and women were asked to return annually.  At each visit, blood was collected between 8:00 and 10:00 AM after a 12-hour fast, during the early follicular phase (cycle days 2–5) whenever possible, and serum was stored at -80F.  In all women who had a natural (non-surgical) MT and a dateable FMP, serum level of AMH was measured from frozen blood samples using a new high-sensitivity monoclonal ELISA with a detection limit of 2 pg/mL (Pico AMH, Ansh Labs, Webster, TX). BMD in the lumbar spine and femoral neck was measured annually in 5 of the 7 study sites.  In 474 women who had AMH and BMD measurements between 2 and 4 years before the FMP, had a 2nd BMD measurement 3-4 years later, and had not taken any medications that affect bone prior to the 2nd BMD measurement, we examined the ability of AMH level to predict the annualized rate of BMD decline between the two visits (% decline per year). AMH inter-quartile range was [11,146] pg/mL. Median rate of BMD decline was 1.3% per year in the spine and 1.0% per year in the femoral neck. Adjusted for age, BMI, smoking, race/ethnicity, and study site, in multivariable linear regression, each 75% (or four-fold) decrement in AMH level was associated with 0.15% per year faster decline in spine BMD (p<0.001) and 0.13% per year faster decline in femoral neck BMD (p=0.005).  These associations persisted even after additional adjustment for time from FMP and serum levels of estradiol and FSH. In multivariable logistic regression, adjusted for age, BMI, smoking, race/ethnicity, and study site, each four-fold decrement in AMH level was also associated with 18% increase in the odds of faster-than-median decline in spine BMD (p=0.02) and 17% increase in the odds of faster-than-median decline in femoral neck BMD (p=0.02). These findings suggest that serum levels of AMH in women going through the MT can indeed predict the rate of trans-menopausal bone loss, and help identify the women at risk of most loss.  AMH levels appear to provide information about the rate of bone loss beyond that provided by serum levels of estradiol and FSH.


Nothing to Disclose: ASK, AS, SAMB, EWY, GAG, PMS, DM, JSF

OR01-3 26302 3.0000 A Anti-Mullerian Hormone and Prediction of Trans-Menopausal Bone Loss 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 1:15:00 PM OR01 7684 11:45:00 AM Osteoporosis: What You Had, What You Lost, and What You Gain Oral

Namki Hong*1, Beom-Jun Kim2, Chong Hwa Kim3, Ki-Hyun Baek4, Yong-Ki Min5, Deog-Yoon Kim6, Seung Hun Lee2, Jung-Min Koh2, Moo-Il Kang7 and Yumie Rhee1
1Department of Internal Medicine, Severance Hospital, Endocrine Research Institute, Yonsei University College of Medicine, Seoul, Korea, Republic of (South), 2Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea, Republic of (South), 3Department of Internal Medicine, Sejong General Hospital, Bucheon city, 4Division of Endocrinology and Metabolism, Department of Internal Medicine, Yeouido St. Mary’s Hospital, Seoul, Korea, Seoul, Korea, Republic of (South), 5Division of Endocrinology and Metabolism, Department of Internal Medicine, Sungkyunkwan University School of Medicine, Seoul, Korea, Republic of (South), 6Department of Nuclear Medicine, Kyunghee University School of Medicine, Seoul, Korea, Republic of (South), 7Seoul St. Mary’s Hospital, The Catholic University of Korea, College of Medicine, Seoul, Korea, Republic of (South)


Abstract: Although bone mineral density (BMD) and several clinical risk factors (CRF) have been identified to predict the risk of osteoporotic fracture (OF), the ability to predict the risk of OF is still suboptimal. Leucine-rich repeat-containing 17 (LRRc17) was known to act as a negative regulator of receptoractivator of NF-kB ligand (RANKL) induced osteoclast differentiation, and thus to play a protective role in bone metabolism.(1) In this case-control study, we aimed to investigate whether decreased circulating plasma LRRc17 level can serve as an independent and additive risk factor for prevalent OF, even after divided into vertebral fracture (VF) and non-vertebral fracture (non-VF). Among 532 consecutive postmenopausal women without any disease or medications that could affect bone metabolism, 102 cases with OF and 102 age and body mass index (BMI)-matched controls (mean age 63.2 years) were enrolled. Morphological VF (n=49) and non-VF (i.e., forearm, humerus, hip, and pelvis; n=60) were identified by lateral thoracolumbar radiographs and an interviewer-assisted questionnaire, respectively. Plasma LRRc17 level was measured using LRRc17 ELISA kit (MyBioSource, San Diego, USA)and natural log-transformed LRRc17 were used due to right-skewed distribution. BMD was measured at lumbar spine and proximal femur by dual-energy X-ray absorptiometry (Lunar Prodigy, Madison, WI). Median plasma LRRc17 levels were significantly lower in subjects with any prevalent OF (117.5 vs. 197.3 pg/ml, P< 0.001), VF (93.2 vs. 172.4 pg/ml, P = 0.002), and non-VF (124.5 vs. 206.9 pg/ml, P = 0.008) compared to their controls. The prevalence of OF was increased in stepwise fashion from the highest LRRc17 tertile (LRRc17 ≥ 228.5 pg/ml, 33.8%) to the lowest (LRRc17 < 95.6 pg/ml, 63.2%). Each one log unit decrease of LRRc17 was associated with increased risk of prevalent OF (Odds ratio [OR] 1.47 [1.10-1.96], P=0.009) and VF (OR 2.43 [1.39-4.23], P=0.002) but only increased tendency with non-VF (OR 1.29 [0.90-1.85], P=0.162) in multivariable logistic models adjusted for CRF and BMD. Adding the information regarding plasma LRRc17 levels to CRF and BMD significantly improved discrimination of prevalent VF patients according to comparison of area under receiver operating characteristics curves (0.71 [0.60-0.82] to 0.81 [0.71-0.90], P=0.036), category-free net reclassification improvement (NRI 0.79, 0.37-1.21, P <0.001), and integrated discrimination improvement (IDI 0.13, 0.06-0.20, P < 0.001) but the improvement was not significant for discriminating non-VF. Our findings suggest that decreased plasma LRRc17 level may be an independent and additive risk factor for prevalent OF, particularly in spine, rather than non-vertebral skeletal sites.


Nothing to Disclose: NH, BJK, CHK, KHB, YKM, DYK, SHL, JMK, MIK, YR

OR01-4 24599 4.0000 A Low Plasma Leucine-Rich Repeat-Containing 17 (LRRc17) Level Is an Independent and Additive Risk Factor for Prevalent Vertebral Fractures in Postmenopausal Women 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 1:15:00 PM OR01 7684 11:45:00 AM Osteoporosis: What You Had, What You Lost, and What You Gain Oral

Alexis Jamie Feuer*1, Ashley Thai2, Ryan T Demmer2 and Maria G Vogiatzi3
1Weill Cornell Medicine New York Presbyterian, New York, NY, 2Mailman School of Public Health, Columbia University Medical Center, NY, NY, 3Children's Hospital of Philadelphia, Philadelphia, PA


Background: Mounting evidence indicates that the sympathetic nervous system (SNS) plays a critical role in bone remodeling. In animals, SNS activation leads to loss of bone mass via norepinephrine stimulated β-adrenergic signaling. Dopamine, which inhibits norepinephrine release in vivo, also regulates bone remodeling in animals. Epidemiologic data suggests adults using β-adrenergic blocking medications have higher bone mineral density (BMD) and reduced fracture risk. However, the effect of amphetamines on bone has been little studied in adults and children. We suggest that stimulant medications, which release and block re-uptake of dopamine and norepinephrine, may affect bone mass.  Stimulant medications such as methylphenidate and amphetamine are widely used for the treatment of Attention Deficit Hyperactivity Disorder (ADHD), an increasingly prevalent neurodevelopmental disorder that affects approximately 6.4 million children in the U.S. As adolescence and young adulthood are critical periods for bone health, it is important to assess the effects of stimulants on bone mass in this population. 

Objective: To investigate associations between stimulant use and bone mass in pediatric subjects.

Design: Cross-sectional analysis of data from the 2005–2010 National Health and Nutrition Examination Study (NHANES).

Participants: 6489 NHANES participants ages 8–20 years (mean 13.58 + 3.58).

Outcomes: Total femur, femoral neck and lumbar spine bone mineral content (BMC) and density (BMD) assessed via dual-energy X-ray absorptiometry (DXA).

Results: 159 of 6489 subjects used stimulants. Stimulant use was an independent predictor of bone mass after multivariable adjustment for age, gender, height and weight Z score, socioeconomic status, physical activity, cotinine level and race/ethnicity. Lumbar spine BMC was 5.1% lower among stimulant users versus non-users (mean difference 0.704 g, SE ± 0.02 g, p = 0.005). Lumbar spine BMD was 3.9% lower (mean difference 0.037 g/cm2, SE ± 0.001 g/cm2, p = 0.002). Femoral neck BMC was similarly negatively associated with stimulant use while BMD approached significance. Compared to nonusers, femoral neck BMC was 5.3% lower among stimulant users (mean difference 0.242 g, SE ± 0.0093 g, p = 0.009) and BMD was 3.7% lower (mean difference 0.034 g/cm2, SE ± 0.0157 g/cm2, p = 0.08). Subjects treated with stimulants for more than 6 months had lower lumbar spine BMD and BMC than those using for less than six months and non-users.

Conclusion: In this NHANES study, pediatric subjects treated with stimulants had lower DXA measurements of the lumbar spine and femoral neck compared to non-users. As stimulant medications are first-line pharmacotherapies for ADHD, their potential effects on pediatric bone health need to be clarified. These findings support the need for future prospective studies to examine the effects of stimulant use on bone mass in this population.


Disclosure: MGV: Advisory Group Member, Novo Nordisk. Nothing to Disclose: AJF, AT, RTD

OR01-5 25830 5.0000 A Use of Stimulant Medications and Bone Mass in Children and Adolescents: An Nhanes Study 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 1:15:00 PM OR01 7684 11:45:00 AM Osteoporosis: What You Had, What You Lost, and What You Gain Oral

Hanh Nguyen1, Phillip Wong2, Boyd J Strauss3, Peter R Ebeling4, Frances Milat5 and Amanda Vincent*6
1Monash University, Clayton, Australia, 2Hudson Institute of Medical Research, 3Monash University, Clayton VIC, Australia, 4Monash University, Australia, 5Hudson Institute of Medical Research, Clayton, Australia, 6Monash Health, Clayton, Victoria, Australia


Background: Turner Syndrome (TS) is the most common female chromosomal abnormality, and results from complete or partial X chromosome monosomy. It is associated with short stature, gonadal failure, osteoporosis, and fragility fractures. Chronic estrogen deficiency leading to suboptimal peak bone mass accrual is a major modifiable risk factor for osteoporosis in TS. Pubertal induction with estrogen replacement therapy (ERT) before age 13 is therefore recommended (1).

Aim: To investigate the prevalence and risk factors for low BMD in an established TS cohort.

Method: A retrospective, cross-sectional study of TS patients who underwent DXA scanning at Monash Health, Victoria, Australia from 1998 to 2015. Cases were identified through the hospital Adult TS clinic and Bone Density Department databases. Data from baseline DXA included areal bone mineral density (aBMD) and Z-scores. To adjust for smaller bone size in TS patients, bone mineral apparent density (BMAD) of the lumbar spine (2) and femoral neck (3) was calculated. Low bone mass in young pre-menopausal women was defined as a Z-score < -2.0 (4). Medical history, including ERT and growth hormone therapy (GHT), was collected from medical records. Statistical analysis included descriptive statistics and multivariate regression analysis.

Results: Seventy-nine TS patients were included, and fractures occurred in 30.4% of cases. The median age of TS diagnosis was 11 (range 0-65) years. Primary amenorrhoea was common (81.8%), and the median age of commencing estrogen was delayed at 16 (range 11-46) years. Non-continuous ERT use was reported in 37.9% and GHT was reported in 53%. The median (range) age, height and body mass index was 29 (14-66) years, 148.5 (127.8 – 170.0) cm, and 25.6 (12.4 – 49.5) kg/m2, respectively. The most common osteoporotic risk factor was gonadal failure (89.4%), followed by vitamin D deficiency (40%). Low bone mass in the spine and femoral neck occurred in 26.4% and 7.7%, respectively. Multivariate regression analysis demonstrated that spine and hip aBMD, and BMAD, were inversely associated with age of commencing ERT or years of estrogen deficiency (adjusting for age at DXA and BMI). After adjusting for confounding, GHT was associated with neither aBMD nor BMAD.

Conclusion: Low spinal bone mass is common in TS. Both the delay in commencing estrogen and years of estrogen deficiency are important independent risk factors for reductions in spine and hip bone mass. Avoiding estrogen deficiency is important in optimizing bone health in TS. This depends on early diagnosis of TS to allow age-appropriate pubertal induction and to maximize ERT compliance. Transitional multidisciplinary TS clinics have a role in the early initiation of ERT and in monitoring bone health in TS.


Nothing to Disclose: HN, PW, BJS, PRE, FM, AV

OR01-6 24834 6.0000 A Delay in Oestrogen Commencement Is Associated with Lower Spine and Hip Bone Mineral Density in Turner Syndrome 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 1:15:00 PM OR01 7684 11:45:00 AM Osteoporosis: What You Had, What You Lost, and What You Gain Oral

Timothy CR Prickett*, Brian A Darlow, Richard Troughton, John Elliott, John Horwood, Julia Martin and Eric A. Espiner
University of Otago, Christchurch, New Zealand


Background: Subjects born with very low birth weight (VLBW) have increased risk of metabolic syndrome and cardiovascular disorders in later life. Raised plasma concentrations of natriuretic peptides are recognized markers of later vascular disease but no study has evaluated their use in young adults born with VLBW. Accordingly we have measured amino terminal proBNP (NTproBNP, a marker of cardiac stress) and amino terminal proCNP (NTproCNP, a putative marker of vascular stress) along with conventional vascular risk factors (VRF) in a cohort of affected VLBW subjects (<1500g) studied sequentially since birth (1). A control group born the same year (1986) with normal birth weight, and matched for gender and ethnicity, was also studied concurrently.

Methods:VLBW adults (135 subjects) and controls (69 subjects), mean age 27.2 yr, were assessed (demographics, clinical and anthropometrics)  over a 2 day period at one centre (Christchurch, New Zealand) during which metabolic studies were performed. Fasting overnight blood samples were drawn for assay of NTproBNP and NTproCNP along with other VRF including cholesterol/ HDL cholesterol ratio, triglycerides, and insulin-glucose ratio. Spearman coefficients were used to determine univariate associations.

Results: VLBW adults were significantly shorter (males 4.1 cm, females 3.9 cm) than controls (p<0.05 and <0.01 respectively). Systolic blood pressure was significantly increased in males (122±1.5 vs 116±2.1 mmHg, p<0.04) but not in females. Body Mass Index (26.4±0.5, VLBW; 27.7±0.6 controls) did not differ. Compared to controls, plasma NTproCNP was higher in male (23.4±0.4 vs 17.2±0.5 pmol/L, p<0.005) and female (18.7±0.4 vs 17.2±0.5 pmol/L, p<0.05) VLBW subjects whereas NTproBNP and other VRF did not differ between the two groups. Across all participants (n=204), NTproCNP was significantly associated with systolic (r=0.33) and diastolic blood pressure (r=0.19), chol/HDL ratio (r=0.21), triglycerides (r=0.24) and insulin resistance (r=0.19). Unexpectedly, associations were all significantly inverse with NTproBNP (range of r values -0.21 to -0.38). Across all subjects, the ratio of NTproCNP to NTproBNP was correlated with a composite index of vascular risk (r=0.46), stronger than the correlation of either NTproCNP (r=0.29) or NTproBNP alone (r=-0.39).

Conclusion: Alone among VRF, and as early as the 3rd decade, plasma NTproCNP is significantly raised in both males and females born with VLBW. The differential association of NTproCNP (positive) and NTproBNP (inverse) with VRF in this young adult population likely reflects an adaptive response to vascular stress (increasing CNP) and genetic polymorphisms raising BNP (2) and conferring reduced risk of future metabolic (3) and vascular complications (4). The ratio of NTproCNP to NTproBNP could be a novel index of metabolic vascular health in young adults.


Nothing to Disclose: TCP, BAD, RT, JE, JH, JM, EAE

OR03-1 25450 1.0000 A New Insights into Cardiac and Vascular Natriuretic Peptides: Findings from Young Adults Born with Very Low Birth Weight 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 1:15:00 PM OR03 7690 11:45:00 AM Cardiometabolic Risk - Salt, Fat and Sex Oral

Katherine Neubecker Bachmann*1, Karen K. Miller2, Thomas J Wang3 and Joel S Finkelstein4
1Vanderbilt University Medical Center, Nashville, TN, 2Massachusetts General Hospital/Harvard Medical School, Boston, MA, 3Vanderbilt University Medical Center, 4Massachusetts General Hospital, Boston, MA


Background: The natriuretic peptide (NP) hormonal system is an important determinant of blood pressure. Low NP levels are associated with hypertension and adverse cardiac remodeling. NP levels are much lower in healthy men compared to healthy women, suggesting that a relative NP deficiency in men might contribute to the higher risk of hypertension and cardiovascular disease observed in men. Prior observational studies suggest an inverse association between testosterone and N-terminal proBNP (BNP) in both sexes. Thus, we tested the hypothesis that testosterone supplementation reduces circulating BNP in men.

Methods: We studied 362 healthy men (mean age 33 years) who were enrolled into one of 3 cohorts: 1) placebo GnRH agonist + placebo testosterone gel (controls, n=35); 2) GnRH agonist (goserelin acetate 3.6 mg monthly) + randomization to placebo gel or testosterone gel 1.25 g, 2.5 g, 5 g, or 10 g daily (n=167), and 3) GnRH agonist (goserelin acetate 3.6 mg monthly) + aromatase inhibitor (anastrazole 1 mg daily) + randomization to placebo gel or testosterone gel 1.25 g, 2.5 g, 5 g, or 10 g daily (n=160). At 12 weeks, we analyzed serum BNP, serum total testosterone, and serum estradiol levels.

Results: As expected, mean levels of serum total testosterone and estradiol differed between testosterone dosage groups (p<0.0001), with higher levels in higher dosage groups. The range of estradiol levels was much narrower in the cohort that received an aromatase inhibitor compared to the cohorts that did not receive an aromatase inhibitor. BNP levels were negatively associated with serum total testosterone levels in the cohort that received an aromatase inhibitor (r = -0.21, p<0.009), in the cohorts that did not receive an aromatase inhibitor (r = -0.14, p<0.05), and in all cohorts combined (r = -0.17, p=0.001); differences remained significant after adjusting for age and BMI. In contrast, BNP levels were not associated with estradiol levels. In multivariable models including serum total testosterone, estradiol, age and BMI, BNP levels were associated negatively with serum total testosterone levels (partial r = -0.19, p=0.0005) but were not associated with estradiol levels.

Conclusions: In men randomized to receive varying doses of testosterone supplementation, there is an inverse association between circulating testosterone and BNP levels.  In contrast, there is no clear association between estradiol and BNP, within the range of estradiol levels seen in men. Further investigation is needed to elucidate the mechanisms underlying the sex-specific differences in NP levels, a topic that has potentially important implications for sex-related disparities in hypertension and other cardiovascular disorders.


Nothing to Disclose: KNB, KKM, TJW, JSF

OR03-2 24755 2.0000 A The Effect of Gonadal Steroids on the Natriuretic Peptide System 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 1:15:00 PM OR03 7690 11:45:00 AM Cardiometabolic Risk - Salt, Fat and Sex Oral

Kenneth Earle*1, Lauren Ng1 and Karima Zitouni2
1St George's University of London, 2St George's University of London, London, United Kingdom


Chronic kidney disease (CKD) is an independent risk factor for hospitalization, and is associated with premature death from cardiovascular causes (CVD).  Progression to end-stage renal disease is higher in non-Caucasian than Caucasian patients with diabetes but the risk of death from CVD is greater in women than in men. Vascular stiffness varies with sex and could promote renal dysfunction, but it is unclear how this relates to CKD and whether there is variance according to heritage. We studied 164 patients without clinical evidence of CVD of Caucasian and non-Caucasian heritage. Infra-red finger plethysmography was used to determine pulse-wave amplitude from which a stiffness index (SI) was computed from the mean of three readings. Body mass index, bioimpedance, sitting blood pressure, glycated haemoglobin, estimated glomerular filtration (eGFR) - using the CKD-EPI equation, and lipids were measured in the fasted state. Data were analysed using parametric and non-parametric tests according to their distribution (Stata 14 Chicago, US). Regression analysis was performed with SI as the dependent variable for gender and for heritage groups.  The cohort was composed of 84 women and 80 men with a similar mean [SD] age, body mass index, sitting systolic blood pressure and HbA1c (61.2[7.0] vs 60.0[8.2] yrs; 31.0[7.0] vs 29.4[6.0] kg/m2; 138.4[19.3] vs 139.8[16.5] mmHg and 59.0[18.0] vs 55.6[17.7] mmol/mol respectively). Vascular stiffness was significantly higher in the Caucasian than in the non-Caucasian heritage groups (11.15[6.57] vs 8.59[4.91] mS; p=0.0065); in the gender sub-groups, bioimpedance was higher in women compared with men (41.7[6.3] vs 29.2[7.3]; p<0.01 and 38.9[8.6] vs 28.9[8.7];p<0.01) but eGFR was similar (89.7[20.2] vs 90.3[16.1] and 86.8[15.7] vs 89.9[17.1] mls/min/173m2).  A linear regression model with SI as the outcome variable including age, blood pressure, HbA1c, eGFR and bioimpedance was significant in women (R-squared =0.20; p=0.026).  There was a positive correlation with bioimpedance (0.309; p=0.007) and negative correlation with eGFR (-0.172; p=0.004). There was no correlation between SI and any of the anthropometric, clinical or biochemical measures in men in either heritage group in this cohort. These data suggest that vascular stiffness has a strong relationsship with measures of body adiposity in women at risk of progressive renal disease in type 2 diabetes. In women of Caucasian heritage, vascular stiffness was associated with lower levels of renal function. The causes of gender differences in vascular stiffness, whether it is causative of renal dysfunction and is preventable by weight management remains to be determined.


Nothing to Disclose: KE, LN, KZ

OR03-3 26146 3.0000 A Sex Differences in Vascular Stiffness and Relationship to the Risk of Renal Functional Decline in Patients with Type 2 Diabetes 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 1:15:00 PM OR03 7690 11:45:00 AM Cardiometabolic Risk - Salt, Fat and Sex Oral

Gianluca Iacobellis* and Mahshid Mohseni
University of Miami, Miami, FL


Epicardial adipose tissue (EAT) is a unique visceral fat deposit with peculiar features and interesting applications. EAT can be measured using standard echocardiography and its thickness is a reliable marker of visceral fat and an emerging therapeutic target. EAT is higher in subjects with type 2 diabetes when compared to controls. Liraglutide, an analogue of glucagon-like peptide-1 (GLP-1RA), improves glycemic control and causes weight loss in type 2 diabetic patients.  Whether Liraglutide-related weight loss could be attributed to a visceral fat loss is unknown and unexplored. We sought to test the hypothesis that Liraglutide causes a rapid and significant reduction in EAT in well controlled overweight and obese type 2 diabetic patients. This was an interventional case-controlled study in type 2 diabetic subjects with body mass index (BMI) > 27 kg/m2 and HemoglobinA1c (HbA1c) ≤ 8% on metformin monotherapy. Individuals were randomized into 2 groups. One group received additional Liraglutide up to 1.8 mg SC once daily and the other group remained on Metformin up to 1 g twice daily. EAT thickness was measured at baseline and after 12 weeks by standard echocardiogram. Currently, 63 patients have completed the 12-week follow upIn the Liraglutide group, EAT decreased from 9.9±2 mm to 6.9±1.8 mm (p< 0.001) after 12 weeks accounting for a 31% reduction. There was no significant reduction in EAT in the Metformin group; Left Ventricular Mass (LVM) decreased from 36.4 to 31.0 g/h2.7 (p<0.01) in the Liraglutide group. BMI decreased from 36.7±6 kg/m2 to 34.5±5 kg/m2 and HbA1c lowered from 6.7±0.7 % to 6.1±0.8 % in the Liraglutide group, reflecting a 6% and 10% reduction, respectively.  Our preliminary results indicate that Liraglutide induces a massive and rapid EAT reduction and cardiac improvement in overweight and obese type 2 diabetic patients, independent of overall weight loss and improved glycemic control. GLP-1RA effects may be therefore visceral fat specific. These data are of great novelty and immediate clinical application.


Nothing to Disclose: GI, MM

OR03-4 25263 4.0000 A Liraglutide Induces a Rapid and Large Epicardial Fat and Left Ventricular Mass Reduction Beyond Weight Loss and Glycemic Control 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 1:15:00 PM OR03 7690 11:45:00 AM Cardiometabolic Risk - Salt, Fat and Sex Oral

Mark Daniel DeBoer*1, Matthew James Gurka2, Jessica G Woo3 and John A Morrison3
1Univ of Virginia, Charlottesville, VA, 2University of Florida, Gainesville, FL, 3Cincinnati Children's Hospital Medical Center, Cincinnati, OH


Background: Adiponectin is secreted by adipocytes in inverse proportion to the amount of fat tissue they contain.  Adiponectin has been implicated as being in the causal pathway of insulin resistance, with low levels being associated with insulin resistance and the metabolic syndrome (MetS).  In longitudinal studies, levels of adiponectin have been inversely associated with risk for future Type 2 diabetes (T2DM) and cardiovascular disease (CVD), while and fasting insulin levels are positively associated with these risks.  We recently reported that the severity of MetS, as determined using a sex- and race/ethnicity-specific MetS severity Z-score, was positively related to future incidence of T2DM and CVD. However, it is unclear what the relationships are between MetS severity and levels of fasting insulin and adiponectin as related to risk for T2DM and CVD.

Methods: We evaluated data from 711 participants of the Princeton Lipid Research Cohort with information regarding levels of insulin, adiponectin, and MetS severity during the time period 1998-2003 (mean age 39.5 years), 595 of whom with data regarding MetS severity in childhood from 1973-1976 (mean age 12.9 years), and 417 with disease status from 2010-2014 (mean age 50.9 years).  We used linear regression to assess relationships between childhood MetS severity Z-score and adult insulin and adiponectin levels; we used Pearson correlation to assess relationships between current levels. Finally, we used logistic regression to assess individual and independent relationships between insulin, adiponectin and MetS severity Z-score on future T2DM and CVD.   

Results: Childhood MetS severity Z-score exhibited positive association with adult levels of insulin (p<0.01) and inverse association with adult levels of adiponectin (p<0.01). Current MetS severity Z-score was linked to fasting insulin (Pearson’s r=0.618, p<0.0001) and adiponectin (r=-0.467, p<0.0001). In individual analyses, higher levels of insulin and MetS severity Z-score as adults (mean age 39.5 years) were related to future odds of incident T2DM and CVD over the next 11.2 years (all p<0.001), while lower levels of adiponectin were only related to odds of future T2DM (p<0.0001). In a model that included insulin, adiponectin and MetS severity Z-score, adiponectin was not linked to either future diabetes or CVD; both insulin (p<0.05) and MetS severity Z-score (p<0.01) were related to risk of future T2DM, while only MetS Z-score was related to future CVD (p<0.001).

Conclusion: The severity of MetS exhibits tight links with adiponectin and fasting insulin, including between childhood and adulthood.  Both fasting insulin and MetS severity Z-score were related to future T2DM independent of adiponectin; of these 3 measures, only MetS severity Z-score was an independent predictor of future CVD.  These data suggest potential clinical utility in assessing MetS severity as a marker of risk.


Nothing to Disclose: MDD, MJG, JGW, JAM

OR03-5 26081 5.0000 A Inter-Relationships Between the Severity of Metabolic Syndrome, Insulin and Adiponectin and Their Relationship to Future Diabetes and Cardiovascular Disease: The Princeton Lipid Research Study 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 1:15:00 PM OR03 7690 11:45:00 AM Cardiometabolic Risk - Salt, Fat and Sex Oral

Ritu Madan*1, Elaine Cochran2, Brent S. Abel2, Ahmed M. Gharib3, Ranganath Muniyappa4 and Rebecca J. Brown5
1U Nebraska MC, 2DEOB, NIDDK, NIH, 3NIDDK, NIH, Bethesda, MD, 4Diabetes Endocrinology and Obesity Branch, NIDDK, NIH, 5NIDDK, NIH


Introduction: Cardiovascular (CV) disease is frequently associated with insulin resistance. Lipodystrophies (LD) are a rare group of disorders characterized by extreme insulin resistance, hypertriglyceridemia, and low HDL. CV disease has been reported as a major cause of morbidity and mortality in LD based on case reports. In this study, we examine the extent of atherosclerotic coronary artery disease in patients with LD using coronary CT angiography compared to age- and sex-matched controls.

Methods: Patients with partial and generalized LD participating in IRB-approved studies at the National Institutes of Health underwent onetime non-contrast cardiac CT for coronary calcium score (Agatston) followed by a standard cardiac multidimensional CT angiogram (CCTA). Patients with LD (n=19; 14 male and 5 female) were compared with age-, sex-, and BMI-matched controls.

From CCTA, the following scores were calculated to quantify atherosclerosis- 1) segment-involvement score (SIS), 2) segment-stenosis score (SSS). The coronary circulation was divided into 16 segments. The SIS (on a scale of 0-16, with 16 the most involved) was calculated based on the number of coronary artery segments involved with any atherosclerotic plaque. Each of the involved coronary artery segments were graded based on the degree of stenoses (from 0-3). The SSS was derived by addition of stenosis score of each individual coronary artery segment and thus ranged from 0-48. Previous studies have shown that subjects with SIS and SSS greater than 5 have higher mortality compared to those with scores below 5.

Results: Age (33.8±13.0 vs. 33.6 ±12.6 yrs) and BMI (24.2±5.2 and 26.8±5.4 kg/m2) were not significantly different between LD and control groups. In the LD group, 36.8% had generalized LD and the remainder had partial LD. As expected, more patients with LD were diabetic (89.4% vs. 4.7%, p<0.001), hypertensive (52.6% vs. 15.7%, p=0.03), and had higher triglycerides (1322 ± 2935 vs. 104± 70 mg/dL, p= 0.0001). Despite these classical CV risk factors, only 57.8% of subjects with LD and 42.1% of controls had SIS > 5 (p=0.35). Similarly, the proportion of subjects with SSS >5 was not different between the two groups (21% vs. 15.7%). Coronary calcium scores also did not differ significantly between the two groups though the number of subjects with calcium scores greater than 0 tended to be higher in LD vs controls (63.1% vs. 21%, p=0.07).

Conclusion: Contrary to our hypothesis, these findings suggest that despite the presence of insulin resistance and other CV risk factors, young patients with LD do not have increased coronary atherosclerotic burden.


Nothing to Disclose: RM, EC, BSA, AMG, RM, RJB

OR03-6 26558 6.0000 A Subclinical Atherosclerosis and Plaque Burden in Patients with Lipodystrophy 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 1:15:00 PM OR03 7690 11:45:00 AM Cardiometabolic Risk - Salt, Fat and Sex Oral

Laya Ekhlaspour*1, Courtney Balliro2, Firas H. El-Khatib3, Debbie Mondesir4, Manasi Sinha2, Kendra L Magyar2, Mallory Hillard5, Lisa Dao6 and Steven Jon Russell2
1Massachusett General Hospital, Boston, MA, 2Massachusetts General Hospital, Boston, MA, 3Boston University, Boston, MA, 4Massachusett General Hospital, Boston, 5Massachusetts General Hospital, 6University of California, Riverside - School of Medicine


Introduction: Tight glycemic regulation is important to avoid complication from diabetes, but it is difficult to achieve without hypoglycemia. One approach to this problem has been development of insulin pumps that use information from a continuous glucose monitor (CGM) to stop insulin delivery at a user-defined threshold. Effectiveness is limited by slow absorption of insulin and the continued absorption of already-delivered insulin after the infusion is stopped. An alternative approach is a pump that automatically gives micro-dose glucagon to prevent hypoglycemia.

Material and Methods: We conducted a double-blinded, randomized, placebo controlled crossover study involving 22 adult subjects with type 1 diabetes who used an insulin pump or multiple daily injections, had a self-reported average frequency of hypoglycemia <60 mg/dl of at least twice a week, and reported inconsistent or absent symptoms with blood glucose <50 mg/dl. Participants administered their own insulin as usual while receiving either glucagon or placebo (filled from coded vial blinding devices) for 24 hours at a time from an automated bionic pancreas system. During the 2-week study, days were randomized to glucagon or placebo in blocks of two (7 days of each). The primary outcome was area over the curve <60 mg/dl (AOC<60), a measure of total hypoglycemia exposure, on glucagon vs. placebo days. Secondary outcomes included AOC<60 during the nighttime (11:00 PM – 7:00 AM), time <60 mg/dl, incidents of symptomatic hypoglycemia, mean CGM glucose, mean glucagon dosing on glucagon days, and daily self-reported nausea on a visual analog scale.

Results: The AOC <60 mg/dl was reduced by 75% on glucagon vs. placebo days (851 ± 748 vs. 3,414 ± 2,242 mg/dl·min, p<0.001). There was a 91% reduction in AOC<60 at night on glucagon vs. placebo days (117 ± 204 vs. 1,309 ± 1,476 mg/dl·min, p<0.0001). Subjects spent 74% less time with BG <60 mg/dL on glucagon vs. placebo days (1.2 ± 0.8% versus 4.7 ± 3.6%; p<0.0001). There were half as many symptomatic hypoglycemia episodes on glucagon vs. placebo days (0.6 ± 0.4 versus 1.2 ± 0.8 incidents per day; p<0.0001). Blinding was effective, with subjects correctly guessing their daily assignment to glucagon or placebo in a daily survey on 42% of days (similar to chance). There was no difference in mean CGM glucose on glucagon vs. placebo days (153 ± 28 vs. 152 ± 27 mg/dl, p=0.6). The mean total daily dose of glucagon was 0.48 mg on glucagon days. There was a trend for more self-reported nausea on glucagon vs. placebo days, but this did not reach statistical significance (1.1 ± 0.6 vs. 0.4 ± 0.7 cm on a 10 cm scale, p=0.05). There was no unexpected or severe adverse events on either glucagon or placebo days.

Conclusions: Automated glucagon administration effectively reduced hypoglycemia in patients with type 1 diabetes, and was well tolerated.


Disclosure: SJR: Scientific Board Member, Tandem Diabetes Care, Ad Hoc Consultant, Sanofi, Speaker, Eli Lilly & Company, Principal Investigator, Eli Lilly & Company, Principal Investigator, Tandem Diabetes Care, Speaker, Dexcom, Speaker, Sanofi, Principal Investigator, Dexcom, Scientific Board Member, Companion Medical. Nothing to Disclose: LE, CB, FHE, DM, MS, KLM, MH, LD

OR12-1 25429 1.0000 A Closed-Loop Glucagon Administration for the Automated Prevention and Treatment of Hypoglycemia in Type 1 Diabetes 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 1:15:00 PM OR12 7705 11:45:00 AM Fresh Thoughts on Diabetes Treatment Oral

Sushela S Chaidarun*1, Kerrington D. Smith1, David A. Axelrod1, Avin P. Pothuloori1, Saumya Saini1, Muriel E. Ward1, Evelyn L. Hanscom1, Samuel J. Kesseli2 and Timothy B. Gardner1
1Dartmouth-Hitchcock Medical Center, Lebanon, NH, 2The Geisel School of Medicine at Dartmouth, Lebanon, NH


Background: Post-pancreatectomy diabetes management is challenging due to its brittle nature. The advent of islet auto-transplantation can help transition patients from post-surgical insulin dependent diabetes to type 2-like diabetes, pre-diabetes, or in some cases, no diabetes at all. However, the procedure is mostly limited to sites with islet isolation facilities. Here we described our experience at Dartmouth-Hitchcock Medical Center, where we initially used off-site and then transitioned to intra-operative islet isolation for patients with severe medically refractory chronic pancreatitis with preserved islet function.

Results: In 2012 Dartmouth-Hitchcock Medical Center developed a collaborative program for total pancreatectomy with islet auto-transplant (TP-IAT), initially using off-site islet isolation at Massachusetts General Hospital and then transitioned to on-site intra-operative islet isolation. We have successfully treated 30 patients thus far, and have had encouraging results from a diabetes perspective. Before the surgery, 11% of the patients already had diabetes requiring treatment (metformin and/or insulin), 15% had pre-diabetes, and 74% had normal A1c <5.6%. The average A1c was 5.7+1.1% (+SD) with a range of 4.6-8.4%. All patients had normal pre-operative stimulated c-peptide (3.2+ 2.1 ng/ml, normal range 1.1-4.4 ng/ml), suggestive of preserved endogenous insulin production. Average islet equivalent (IEq) yields was 4,683 IEq/Kg (range 843-10,214); 10% of patients had low yields (< 2,500), 57% had moderate yields (2,500-5,000), and 33% had high yields (>5,000 IEq/Kg). Three months after the surgery, 40% of patients required no insulin with near normal mean A1c 6.3% while 83% had A1c at target (<7%). Six months post-operatively, the average A1c was 6.8% while 71% of patients had A1c at target. After 12 months, the average A1c had risen to 7.5%, but 62% of the patients still had glycemic control at target, and 11 of 21 (52%) of patients required no insulin at all. Since most of the patients maintained reasonable islet graft function with normal c-peptide levels, we typically started treatment with oral agents (e.g. metformin and then DPP-4 inhibitor) without the need of insulin for patients who subsequently had suboptimal diabetic control.

Conclusion: TP-IAT techniques, either using off-site or on-site intra-operative islet isolation, resulted in a lower rate of insulin-dependent diabetes, and should be seen as a viable treatment option for patients with intractable chronic pancreatitis. Furthermore, patients generally experienced much less pain and returned to work with a better quality of life. Therefore, islet auto-transplantation either on-site or off-site islet isolation can be used to avert the burden of complicated and costly diabetic care for carefully selected patients requiring total or sub-total pancreatectomy.


Nothing to Disclose: SSC, KDS, DAA, APP, SS, MEW, ELH, SJK, TBG

OR12-2 24190 2.0000 A Diabetes Management and Outcomes of Islet Auto-Transplant after Total Pancreatectomy: Dartmouth Experience, Using Off-Site and Intra-Operative Islet Isolation 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 1:15:00 PM OR12 7705 11:45:00 AM Fresh Thoughts on Diabetes Treatment Oral

Daniela Jakubowicz*1, Zohar Landau2, Julio Wainstein3, Yosefa Bar-Dayan4 and Oren Froy5
1Tel Aviv University, Tel Aviv, Israel, 2Wolfson Med Ctr, Holon, Holon, Israel, 3E. Wolfson Medical Center. Tel Aviv University, Holon, Israel, 4Tel Aviv University, Israel, Holon, Israel, 5The Hebrew University of Jerusalem, Rehovot, Israel


Background: Substantial evidence supports that a diet with high caloric protein breakfast (B) and reduced dinner (D) is a successful strategy for weight loss (WL), increased overall satiety (S) and reduced overall postprandial glycemia (PPG) and HbA1c in obese and type 2 diabetes (T2D). Particularly whey protein exerts potent direct insulinotropic  effect and through stimulation of glucagon-like peptide 1 (GLP-1) leading to reduction of PPG in T2D,

Objective: To search whether high protein B consisting of whey protein has a greater impact on WL, overall PPG, S and HbA1c than B with different protein sources.

Methods: 48 T2D, 22 males, BMI 32.1 0.9 kg/m2, aged 58.9 4.5 y, were randomized to 3 isocaloric (1500 kcal) WL diets during 12 weeks. The 3 diets had breakfast (660 kcal), lunch (567 kcal) and dinner (276 kcal) with the same composition at lunch (% of carb:prot:fat 20:45:25%) and dinner (13:40:47%), but differed in the B composition and protein content.

The 3 meals were: 1) A high-carbohydrate B Diet (CBd), n=15, with 13 g protein at B: (65:15:20%), i.e., ready-to-eat cereals. 2) A high-protein B Diet (PBd), n=16, with 36 g protein in B: (40:40:20%), i.e., eggs, tuna and cheese. 3) A whey B Diet (WBd), n=17, with 36 g protein in B, (40:40:20%), i.e., whey protein shake. Additionally, on 3 separate days between day 10 and 15 after diet initiation, all participants underwent 3 all day CBd, PBd and WBd meal tests, to assess PPG, insulin, intact GLP-1 and S after B, lunch and D.

Results: After 12 weeks, WL was -3.5 ± 0.3 kg (-3.8%) in CBd, -6.1 ± 0.3 kg (- 6.8 %) in PBd and the greater WL was found in WBd, -7.6 ± 0.3 kg (- 8.4%)  (p<0.0001). Compared to CBd, the % of WL in PBd was higher by 44 %, and in WBd was greater by 55% (p<0.0001).

The reduction of HbA1c was lower in CBd by -0.36 ± 0.04%, in PBd was -0.6 ± 0.04% and the greater reduction was found in WBd, 0.89 ± 0.05% (p<0.0001). The % of change for HbA1c was 4.6 % in CBd, 7.7% in PBd and the greatest reduction changes were in WBd by 11.5% (p<0.0001). Compared to CBd the % of the reduction of HbA1c was greater by 41 % in PBd, and by 64 % in WBd (p<0.0001).

Overall AUC for PPG was 95522 ± 565 mg/dl*min in CBd, 84065 ± 299 mg/dl*min in PBd and 77452 ± 292 mg/dl*min in WBd (p<0.0001). Compared to CBd the AUC for overall PPG was 12 % lower in PBd and 19 % lower in WBd (p<0.0001). The AUC for overall Insulin was 27% higher in PBd and 38% higher in WBd (p<0.0001). The AUC for overall intact GLP-1 increased more than in CBd, by 32% in PBd and by 41% in WBd (p<0.0001) and AUC for overall S was enhanced by 24% in PBd and by 30% in WBd compared to CBd (p<0.0001).

Conclusions: This study demonstrates that increasing protein content from 13 g to 36 g at breakfast has a significant impact on WL, overall S, HbA1c and overall PPG. However, for the same protein content, whey protein vs other protein sources, yields additional benefits on WL, overall S, reduction of PPG and HbA1c. Whey protein should be considered an important adjuvant in the management of T2D


Nothing to Disclose: DJ, ZL, JW, YB, OF

OR12-3 25019 3.0000 A Whey Protein  Induces Greater Reduction of Postprandial Glycemia and HbA1c, Weight Loss and Satiety Compared to Other Protein Sources  in Type 2 Diabetes 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 1:15:00 PM OR12 7705 11:45:00 AM Fresh Thoughts on Diabetes Treatment Oral

Alessandra Celli*1, Dean Blevin1, Yoan Barnouin1, Georgia Colleluori1, Reina Condevillamar Villareal2 and Dennis Tan Villareal2
1Baylor College of Medicine, 2Michael E. DeBakey VA Medical Center


Background: Type 2 diabetes (T2D) is highly prevalent in older adults due to increasing adiposity and physical inactivity with advancing age. In fact, obesity worsens the metabolic and physical complications of aging that impair quality of life. Although lifestyle intervention is the primary therapy for T2D, this treatment approach is controversial in older adults because of potential exacerbation of sarcopenia and frailty. Currently, there are no directly applicable clinical trial data of lifestyle intervention in older adults with T2D, with recommended treatment approaches primarily based on expert opinion rather than high-level evidence.  

Methods/design: We present preliminary results of an ongoing randomized controlled trial (RCT) designed to comprehensively examine the effect of a lifestyle intervention strategy (behavioral diet therapy for weight loss and exercise training) in older (age: 65-85 yrs.) overweight/obese (BMI ≥ 27 kg/m2) adults with T2D. Main outcomes for this RCT included changes in: a) glycemic metabolic control (HbA1c), b) body weight and body composition (lean body mass, fat mass and visceral fat mass using DXA), c) physical function (Physical Performance Test [PPT], aerobic capacity [VO2peak]), and d) bone mineral density (BMD) and bone quality (trabecular bone score, TBS).  Subjects were randomized to intensive lifestyle intervention (LI group) or healthy-lifestyle control (HL group) for 6 months.

Results: To date, 17 subjects (age: 70.2±3.8 yrs., BMI: 35.3±6.0 kg/m2, HbA1c: 7.3±1.2%) have been enrolled and 13 subjects (76.5%) have completed the interventions. There were no significant differences in age and baseline BMI and HbA1c between the LI (n=10) and HL (n=7) groups. After 6 months, HbA1c significantly improved in the LI group compared with HL (-0.7±0.2 vs 0.2±0.4%; p<0.001). Body weight tended to decrease in the LI relative to HL (-8.1±3.2 vs. -1.9±7.7 kg; p=0.08) associated with significant decreases in fat mass (-4.6±6.6 vs. -2.1±6.6 kg, p=0.02) and visceral fat mass (-0.2±0.2 vs 0.0±0.1 kg; p=0.04) but relative preservation of lean body mass (0.3±3.4 vs. 0.9±5.1 kg; p=0.80). In addition, PPT (3.3±1.6 vs. 1.2±6.5) and VO2peak (2.7±1.1 vs. 0.4±1.5) also significantly improved in LI compared to HL. Finally, although there were no significant changes in BMD, TBS significantly improved in the LI group but not in the HL group (0.09±0.0 vs -0.02; p=0.04).  

Conclusion: These preliminary findings suggest that an intensive lifestyle intervention is not only feasible but importantly confers beneficial effects on glucose control, body composition, and physical function in older adults with T2D.  Further, they provide evidence that bone quality may improve independent of changes in BMD in response to lifestyle intervention. Long-term studies involving a larger sample are needed to follow up on these encouraging results and examine underlying mechanisms.


Nothing to Disclose: AC, DB, YB, GC, RCV, DTV

OR12-4 25882 4.0000 A Intensive Lifestyle Intervention in Older Adults with Diabetes Improves Glycemic Control, Body Composition, Physical Function, and Bone Quality 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 1:15:00 PM OR12 7705 11:45:00 AM Fresh Thoughts on Diabetes Treatment Oral

Ana Priscila Soggia*, Marcelo Salem, Ricardo Abdalla, Luciana Neves, Andre Chiodi Elias, Thais P Sickler, Erika Bezerra Parente, Carlos Basualdo, Sandra Siqueira, Amanda Claro, Natalia Ribeiro, Gabrielle Torres, Jose Antonio Marcondes, Denise Duarte Iezzi and Claudia Kalil
Sirio Libanes Hospital, São Paulo, Brazil


Bariatric surgery is performed to treat obesity class II and III with diabetes remission in 80-90% of cases. Recent studies proposed bariatric surgery for treating diabetes in case of obesity class I (BMI 30-35kg/m2). The first choice is the gastric bypass, however, sleeve with ileal transposition is a new technique proposed, but not yet approved, that changes the insulin secretion and resistance without causing major changes in weight and nutritional deficiencies.

In this randomized, nonblinded, single-center trial, to evaluate the effectiveness on glycemic control, diabetes remission, security and weight control we evaluated three treatment arms: clinical, gastric bypass and sleeve with ileal transposition (sleeve-IT); in 42 obese class I patients with uncontrolled type 2 diabetes.

Patients’ average age (±SD) was 51±7 years old, and 62% were women. The average glycated hemoglobin level was 9.3±1.9%. Glycemic control was defined as glycated hemoglobin level of 6.5% or less, that was achieved in 8% (1 of 12 patients) in clinical group versus 46% (6 of 13 patients) in the bypass group and 100% (12 of 12 patients) in the sleeve-TI group (p= 0,002 sleeve vs bypass). Diabetes remission was defined as glycemic control without medication and was achieved in 30% (4 of 13 patients) in the bypass group versus 75% (9 of 12 patients) in sleeve-TI group (p=0,02 sleeve vs bypass). Glycemic control improved in all three groups, with an average glycated hemoglobin level of 8±1.3% in the clinical group, 6.9±1.0% in the bypass group and 5,6±0,6% in the sleeve-TI group (p=0,01 sleeve vs bypass). Weight loss was greater in the bypass and sleeve-TI (−22.5± 8,2kg and −18±6,4 kg, respectively) than in the clinical group (−4,7±5,1 kg) (p=0.003 for bypass and p=0,017 for sleeve-TI versus clinical group). Drugs to lower glucose, lipid, and blood-pressure levels decreased after both surgical groups. Four patients experienced serious adverse event. There were no deaths or life-threatening complications.

Although it has been recommended in 2010, by International Diabetes Federation, surgery in diabetic patients with BMI between 30-35 when clinical treatment is not successful, few studies have been published. This study showed that the sleeve-TI surgery has better results in glycemic control than the clinical and bypass treatment. The protocol follow-up will continue for 24 to 36 months to assess the maintenance of the results (NCT01857076).


Nothing to Disclose: APS, MS, RA, LN, ACE, TPS, EBP, CB, SS, AC, NR, GT, JAM, DDI, CK

OR12-5 26889 5.0000 A Bariatric Bypass Surgery Versus Sleeve with Ileal Transposition Surgery Versus Clinical Treatment in Obese Class I Patients with Diabetes 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 1:15:00 PM OR12 7705 11:45:00 AM Fresh Thoughts on Diabetes Treatment Oral

Tim Heise*1, Kirstine Stender-Petersen2, Ulrike Hövelmann1, Jacob Bonde Jacobsen2, Leszek Nosek1, Eric Zijlstra1 and Hanne Haahr2
1Profil, Neuss, Germany, 2Novo Nordisk A/S, Søborg, Denmark


Faster-acting insulin aspart (faster aspart) is insulin aspart (IAsp) in a new formulation, with faster initial absorption rate following subcutaneous (s.c.) injection. This trial compared the pharmacokinetic (PK) and pharmacodynamic (PD) properties of faster aspart with those of IAsp at three clinically relevant doses in adults with type 1 diabetes (T1D).

Forty-six subjects with T1D (mean age [± SD]: 44.0 ± 10.4 years) received a single dose (0.1, 0.2 or 0.4 U/kg) of faster aspart or IAsp in a randomized, double-blind, crossover treatment sequence. PK/PD properties of faster aspart and IAsp were evaluated in an automated euglycemic glucose clamp setting (ClampArt, blood glucose [BG] target 5.5 mmol/L [100 mg/dL]; duration 12 h post-dose).

Across all three doses, onset of appearance with faster aspart occurred approximately twice as fast (mean [95% CI] and relative differences: 0.1 U/kg, –4.7 min [–6.0; –3.4], 47%; 0.2 U/kg, –4.7 min [–5.4; –4.0], 54%; 0.4 U/kg, –4.1 min [–5.3; –3.0], 56%) as with IAsp. Time to 50% maximum exposure (Cmax) was observed significantly earlier (8–12 min; 24–32%) across all doses with faster aspart, compared with IAsp. Insulin exposure (AUCIAsp, 0–30 min) was higher in the first 30 min with faster aspart (treatment ratio [95% CI]: 0.1 U/kg, 1.54 [1.29; 1.83]; 0.2 U/kg, 2.09 [1.83; 2.38]; 0.4 U/kg, 2.20 [1.85; 2.62]), compared with IAsp. For both faster aspart and IAsp, total exposure (AUCIAsp,0–12 h) and Cmaxincreased with increasing dose levels.

Onset of action (time to first lowering of BG by 0.3 mmol/L [5 mg/dL] from baseline) occurred up to 26% faster with faster aspart (mean [95% CI] and relative differences: 0.1 U/kg, -5.0 min [–10.0; –0.0], 20%; 0.2 U/kg, –5.8 min [–8.6; –3.1], 26%; 0.4 U/kg, –5.6 min [–8.8; –2.4], 26%) than with IAsp. Likewise, time to 50% maximum glucose infusion rate occurred significantly earlier (9–12 min; 22–25%) with faster aspart than with IAsp across all doses. The early glucose-lowering effect for faster aspart was up to twofold greater within the first 30 min (treatment ratio [95% CI]: 0.1 U/kg, 1.48 [1.00; 2.41]; 0.2 U/kg, 1.92 [1.51; 2.58]; 0.4 U/kg, 2.13 [1.60; 3.06]), compared with IAsp. Total and maximum glucose-lowering effects were similar between faster aspart and IAsp. Both treatments were well tolerated and had no safety issues; no injection-site reactions were observed.

Faster aspart showed a faster onset of exposure and a greater initial absorption than IAsp, which was associated with a faster onset and greater early action, compared with IAsp across all three doses. Hence, the fast absorption properties of faster aspart are preserved across a clinically relevant dose range.


Disclosure: TH: travel grant, Mylan, travel grant, Eli Lilly, Speaker Bureau Member, Novo Nordisk, Scientific Board Member, Mylan, Speaker Bureau Member, Eli Lilly, Researcher, Senseonics, Researcher, Sanofi, Researcher, Novartis Pharmaceuticals, Researcher, Novo Nordisk, Researcher, Medtronic, Researcher, Eli Lilly, Researcher, Grünenthal, Researcher, Dance Pharmaceuticals, Researcher, Boehringer Ingelheim, Researcher, Biocon, Researcher, BD, Researcher, Astra Zeneca, Researcher, Adocia, Travel grant, Novo Nordisk, Advisory Group Member, Novo Nordisk. KS: Employee, Novo Nordisk. JB: Employee, Novo Nordisk. EZ: n/a, Novo Nordisk, n/a, Dance Biopharm, Inc. HH: Employee, Novo Nordisk, Stocks, Novo Nordisk. Nothing to Disclose: UH, LN

OR12-6 25845 6.0000 A Higher Early Insulin Exposure and Greater Early Glucose-Lowering Effect with Faster-Acting Insulin Aspart vs Insulin Aspart Across a Clinically Relevant Dose Range 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 1:15:00 PM OR12 7705 11:45:00 AM Fresh Thoughts on Diabetes Treatment Oral

Angela K Lucas-Herald*1, Silvano Bertelloni2, Anders Juul3, Jillian Bryce1, Jipu Jiang1, Martina Rodie1, Massoud Boroujerdi4, Marie Lindhart Johansen5, Olaf Hiort6, Paul-Martin Holterhus7, Martine Cools8, An Desloovere9, Guilherme Guaragna Filho10, Gil Guerra-Junior11, Naomi Weintrob12, Sabine Elisabeth Hannema13, Stenvert L Drop14, Tulay Guran15, Feyza Darendeliler16, Sukran Poyrazoglu17, Anna Nordenstrom18, Ieuan Arwel Hughes19, Carlo L Acerini19, Rieko Tadokoro-Cuccaro20 and S Faisal Ahmed1
1University of Glasgow, Glasgow, United Kingdom, 2University Hospital Pisa, Pisa, Italy., 3University of Copenhagen, Faculty of Health and Medical Sciences, Copenhagen, Denmark, 4University of Glasgow, Glasgow, UK, 5University of Copenhagen, Copenhagen, Denmark, 6University of Luebeck, Luebeck, Germany, Luebeck, Germany, 7Univ Hosp Schleswig-Holstein, Kiel, Germany, 8Univ Hosp Ghent, Ghent, Belgium, 9University Hospital Ghent, Ghent, Belgium, 10Unidade de Endocrinologia Pediátrica, Universidade de Campinas, São Paulo, Brasil, 11State University of Campinas (UNICAMP),SP, Brazil, Campinas - SP, Brazil, 12Tel Aviv Medical Center, Hod Hasharon, Israel, 13Leiden University Medical Centre, Leiden, The Netherlands, 14Leiden University Medical Centre, Leiden, The Netherlands, Rotterdam, Netherlands, 15Marmara University, Istanbul, Turkey, 16Istanbul University, Istanbul, Turkey, 17Istanbul University, Istanbul Faculty of Medicine, Istanbul, Turkey, 18Karolinska Institutet, Stockholm, Sweden, 19University of Cambridge, Cambridge, UK, Cambridge, United Kingdom, 20University of Cambridge, Cambridge, UK


Background: In boys with a suspected disorder of sex development (DSD) due to Partial Androgen Insensitivity Syndrome (PAIS), systematic evidence that supports the value of identifying a mutation in the androgen receptor gene (AR) is lacking.

Objective: To assess the clinical characteristics and long-term outcomes in young men with suspected PAIS.

Methods: Through the I-DSD Registry, information on the initial presentation and current clinical status was gathered on young men diagnosed as having PAIS (n=52) who presented before the age of 16 yrs and who had undergone genetic analysis of AR.

Results: The median age at presentation and at the time of the study was 1 month (1 day-16 years) and 22 yrs (16-52), respectively. Of the cohort, 29 (56%) had 20 different AR mutations reported. Median external masculinisation score (EMS) at time of diagnosis for AR mut+ve and –ve cases was 7 and 6, respectively (p=0.83). Median current EMS was 9 and 10, in the two respective groups (p=0.09). Testosterone therapy was reported in 16 AR mut+ve (53%) and 6 (26%) AR mut-ve cases. Thirty-one (60%) men required at least one surgical procedure, with AR mut+ve men being more likely to require multiple surgeries for hypospadias repair (p=0.004). All AR mut+ve men had gynaecomastia compared to 9% of the AR mut–ve men. Of the 6 men who had mastectomy, 5 (83%) were AR mut+ve.

Conclusions: Boys with PAIS with an identifiable mutation in AR are at increased risk of poorer medical and surgical outcomes in adulthood. Routine genetic analysis of AR in boys with XY DSD will guide prognosis and personalised management.


Nothing to Disclose: AKL, SB, AJ, JB, JJ, MR, MB, MLJ, OH, PMH, MC, AD, GG, GG, NW, SEH, SLD, TG, FD, SP, AN, IAH, CLA, RT, SFA

OR10-1 27136 1.0000 A The Long Term Prognostic Value of Identifying a Mutation in the Androgen Receptor Gene in Boys with Partial Androgen Insensitivity Syndrome 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Reproductive Endocrinology Friday, April 1st 1:15:00 PM OR10 7719 11:45:00 AM Recent Developments in Disorders of Sex Development (DSD) and Transgender Medicine Oral

Nathalia Lisboa Gomes*1, Sorahia Domenice2, Flávia Siqueira Cunha3, Rafael Loch Batista4, Daniela Rodrigues Moraes5, Mirian Y Nishi3, Mariana F A Funari6, Elaine M F Costa7 and Berenice B Mendonca8
1University of São Paulo, Hospital das Clinicas, Sao Paulo, Brazil, 2University of São Paulo, Hospital das Clinicas, São Paulo, Brazil, 3Hospital das Clínicas, Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brazil, 4Universidade de São Paulo, Sao Paulo SP, 5University of São Paulo, Hospital das Clinicas, Brazil, 6Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo, Sao Paulo - SP, Brazil, 7Hospital das Clinicas, Faculdade de Medicina, Universidade de São Paulo, Sao Paulo, Brazil, 8Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo FMUSP, São Paulo, Brazil


Studies on 46,XY partial gonadal dysgenesis (PGD) patients in adulthood are scarce. Our aim was to retrospectively analyze the phenotype and genotype of 25 patients with 46,XY PDG followed until adulthood. The age of the first visit ranged from 10 days to 34 years old and follow-up ranged from 2.7 to 26 years. Molecular diagnosis by Sanger method included  screening of SRY, SF1, WT1, CBX2, MAPK3, FGF9, FGFR2, GATA4 genes for mutation and DAX1 for duplication. Gender assignment at birth was male in 15 patients and female in 9. Two females changed to male social sex at the age of 8 and 17 and one male changed to female social sex at the age of 17. The diagnosis of gonadal dysgenesis was confirmed by the presence of Mullerian derivatives (18 patients) and/or at least one gonad with histological features compatible with testicular dysgenesis (16 cases). In the female final social sex group (n=10), all patients had bilateral cryptorchidism, 6 patients had clitoromegaly and 1 had micropenis; 6 patients had two perineal openings and a 3 had an urogenital sinus. Bilateral gonadectomy were performed in all at 1.2 to 15.2 years old. In the final male social sex group (n=16), all patients had atypical genitalia characterized by hypospadias (9 with proximal hypospadias), microfallus, bilateral cryptorchidism in 9 patients and unilateral in 3 patients. At the first visit, 3 patients were already gonadectomized. In childhood, 6/13 had preserved testosterone secretion (5 had unilateral cryptorchidism and one had topic testes). During the follow-up, 3 patients lost the capacity to secrete testosterone and 3 developed spontaneous puberty. After puberty, 2 of those patients had LH and FSH slightly elevated and testosterone levels at normal adult range. Two patients had germ cell tumor. Molecular diagnosis was possible in 10/25 cases: mutations were found in SRY in two patients, in SF1 in two, in WT1 mutations also in two (both with typical features of Frasier and Denys-Drash syndromes), in MAPK3 and FGFR1 in two sisters, in CBX2.2 in one patient and DAX1 duplication also in one. We concluded that the phenotype of 46,XY PDG patients is quite variable. Most patients had male assignment (64%) but the majority of them were well adapted in both social sexes in adulthood. Few patients reared as male kept testicular function in adulthood. Molecular diagnosis was reached in 40% of the cases by conventional Sanger sequencing for target genes.


Nothing to Disclose: NLG, SD, FSC, RLB, DRM, MYN, MFAF, EMFC, BBM

OR10-2 27048 2.0000 A Phenotype, Genotype and Follow up of 46,XY DSD Patients Due to Partial Gonadal Dysgenesis 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Reproductive Endocrinology Friday, April 1st 1:15:00 PM OR10 7719 11:45:00 AM Recent Developments in Disorders of Sex Development (DSD) and Transgender Medicine Oral

Rafael Loch Batista*1, Marlene Inacio2, Sorahia Domenice3, Flávia Siqueira Cunha4, Nathalia Lisboa Gomes5, Daniela Rodrigues de Moraes6, Vinicius N. Brito4, Elaine M F Costa7 and Berenice B Mendonca8
1Universidade de São Paulo, Sao Paulo SP, 2Universidade de São Paulo, Sao Paulo - SP, Brazil, 3University of São Paulo, Hospital das Clinicas, São Paulo, Brazil, 4Hospital das Clínicas, Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brazil, 5Universidade de São Paulo, Brazil, 6Universidade de São Paulo, 7University of São Paulo, Sao Paulo, Brazil, 8Hospital das Clínicas, Faculdade de Medicina da Universidade de São Paulo, Brazil


Psychosexual Outcomes in Male and Female 46,XY DSD Patients in Adulthood

Rafael Loch Batista, MD; Marlene Inacio, PhD, Sorahia Domenice MD, PhD; Flávia Siqueira Cunha, MD; Nathalia Lisboa, MD; Vinicius Nahime Brito MD, PhD; Elaine Maria Frade Costa, MD, PhD, Berenice Bilharinho de Mendonça, MD, PhD.

Several psychosexual issues are relevant and rarely explored in patients with disorders of sex development (DSD). There are few reports on the psychosexual outcomes of 46,XY DSD and most of the studies are in 46,XX DSD due to CAH. Patients with 46,XY DSD exhibit a variety of phenotypes at birth ranging from typical female genitalia to undervirilized male external genitalia. Changing from female to male social sex is common, especially in 5α-RD2  and in 17β-HSD3 deficiencies. We evaluated the psychosexual outcomes in 140 patients with 46,XY DSD with different etiologies (testosterone synthesis defects (n=33), 5α-RD2 deficiency (n=30), androgen resistance (n=34), defects in gonadal differentiation (n=33) and with indeterminate etiology (n=10) applying a specific questionnaires on sexual issues. Among these patients, 27 (23.9%) changed the social sex in adulthood (24 female to male and 3 male to female). The psychosexual outcomes were compared between the groups that changed social sex vs the group that kept the social sex in adulthood and between the male and female final social sex in adulthood. In male social sex, we did not observe differences between the group that changed with group that kept the male social sex (p >0.05). In male and female social sex, comparing patients who changed social sex versus the ones who kept the assigned sex, we observed differences on the frequency of orgasm (p=0.015), masturbation (0.02), satisfactory intercourse (<0.001) and sexual life satisfaction (0.016). All parameters were better in the male sex. There was better satisfaction with sexual life in males (73%) compared to females (51%), although these rates are lower than that observed in non-DSD control population (1, 2). We conclude that psychosexual outcomes in male and female 46,XY DSD patients in adulthood are adequate but the patients with male social sex shows better parameters.

References: (1) Moreira ED Jr, Abdo CH, Torres EB, et al. Urology 2001 Oct;58(4):583-8. (2) Abdo CH, Oliveira WM Jr, Moreira ED Jr et al. Int J Impot Res. 2004 Apr;16(2):160-6.

Nothing to Disclose: RLB, MI, SD, FSC, NL, VNB, EMFC, BBM


Nothing to Disclose: RLB, MI, SD, FSC, NLG, DRD, VNB, EMFC, BBM

OR10-3 27049 3.0000 A Psychosexual Outcomes in Male and Female 46,XY DSD Patients in Adulthood 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Reproductive Endocrinology Friday, April 1st 1:15:00 PM OR10 7719 11:45:00 AM Recent Developments in Disorders of Sex Development (DSD) and Transgender Medicine Oral

Ariella Weinberg-Shukron*1, Paul Renbaum2, Rachel Kalifa3, Sharon Zeligson4, Ziva Ben-Neriah5, Amatzia Dreifuss6, Amal Abu-Rayyan7, Noa Maatuk8, Nilly Fardian6, Dina Rekler6, Moien Kanaan7, Abraham O. Samson8, Ephrat Levy-Lahad9, Offer Gerlitz6 and David Zangen10
1The Hebrew University Hadassah Medical school, Jerusalem, Israel, 2Shaare Zedek Medical Center, Israel, 3Hebrew University, Faculty of Medicine, Jerusalem, 4Shaare Zedek Medical Center, 5Genetic clinic, Hadassah Medical Center, Jerusalem, Israel, 6Hebrew University, Faculty of Medicine, Jerusalem, Israel, 7Bethlehem University, Bethlehem, Palestine, 8Bar-Ilan University, Safed, Israel, 9Shaare Zedek Medical Center, Jerusalem, Israel, 10Hadassah Hebrew University Medical Center, Jerusalem, Israel


Ovarian development and maintenance are poorly understood. XX-female gonadal dysgenesis (XX-GD) is a rare, genetically heterogeneous disorder that is characterized by underdeveloped, dysfunctional ovaries with subsequent lack of spontaneous pubertal development, primary amenorrhea, and hypergonadotropic hypogonadism. We report an extended consanguineous family of Palestinian origin in which four females exhibited XX-GD. Using homozygosity mapping and whole exome sequencing, we identified a recessive missense mutation in nucleoporin-107 (NUP107, c.1339G>A, p.D447N). This mutation segregated with the XX-GD phenotype and was not present in available databases or in 150 healthy, ethnically matched controls. NUP107 is a component of the nuclear pore complex, and the NUP107-associated protein Seh1 is required for oogenesis in Drosophila. We found that Nup107 RNAi knockdown in Drosophila somatic gonadal cells resulted in female sterility. Transgenic rescue Drosophila females bearing the Nup107D364N mutation, which corresponds to the human NUP107:p.D447N, resulted in almost complete sterility, with a marked reduction in progeny, morphologically aberrant eggshells, and disintegrating egg-chambers, indicating defective oogenesis and resembling the human phenotype. In order to characterize the defects observed in Nup107D364N mutant ovaries, we immunostained the dissected ovarioles for two proteins representing two major pathways involved in gonadogenesis and oogenesis. C(3)G, a protein that forms the transverse filaments of the synaptonemal complex (SC), which is important in meiosis and homologous recombination was mis-localized in Nup107D364N mutant ovaries. While ovarioles from WT rescue flies showed normal localization of C(3)G to the oocyte's nucleus, in mutant Nup107D364N ovarioles C(3)G was distributed throughout the oocyte's cytoplasm. In contrast, the EGF-R ligand Gurken (grk), which directs the establishment of the dorsal-ventral axis of the oocyte and the future embryo in Drosophila, was correctly localized to the dorsal corner in Nup107D364N mutant oocytes. In summary, these results indicate NUP107 as a novel genetic etiology for ovarian mis-development and suggest that nucleoporin defects may play a role also in milder and more common conditions such as premature ovarian failure. Future research will elucidate the signaling pathways involved in NUP107 mediated ovarian development and will be a valuable insight into ovarian development and dysgenesis mechanisms.


Nothing to Disclose: AW, PR, RK, SZ, ZB, AD, AA, NM, NF, DR, MK, AOS, EL, OG, DZ

OR10-4 24492 4.0000 A XX-Disorder of Sexual Development with Ovarian Dysgenesis Is Caused By a Mutation in the Nucleoporin-107 Gene 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Reproductive Endocrinology Friday, April 1st 1:15:00 PM OR10 7719 11:45:00 AM Recent Developments in Disorders of Sex Development (DSD) and Transgender Medicine Oral

Marissa Paige Grotzke*1, Breeze Hannaford2, Melissa Brewster2, Ken Adamson2, Nathan Askerlund2, Tiffany Atkinson2, Misty Guevara2, Brandon Gwilliam2, Joan Hadley2, Cynthia Hudgens2, Cheryl Kaye2, Robin Lines2, Jo Merrill2, Alan Morris2 and Amber Taylor2
1Veterans Health Administration Salt Lake City Health Care System, Salt Lake City, UT, 2Salt Lake City Veterans Affairs Medical Center, Salt Lake City, UT


Introduction: Although a relatively uncommon medical condition, individuals with Gender Identity Disorder (GID) have unique concerns related to health care (1).  Studies show higher rates of GID in the military and its veterans than the general American population, with most recent estimates suggesting a rate of GID in patients in the Veterans Affairs Health Care System (VAHCS) of 22.9/100,000 persons compared to 4.3/100,000 persons in the general population (2,3). High rates of depression and other mental health disorders are documented in veterans; those with GID are particularly at risk (3,4). Notably, estimated rates of suicide-related events among GID veterans are 20 times higher than the general VAHCS population (3). Mental health disorders frequently contribute to decreased compliance with medical therapies, complicating GID management (5).

Objectives: To address these concerns, we recently formed a multi-disciplinary Gender Dysphoria Team with endocrinology, mental health, pharmacy, speech therapy, and vocational rehab providers at the Salt Lake City Veterans Affairs Medical Center (SLC VAMC).  Providers meet with patients individually and bi-monthly as a team to discuss patients and treatment plans. We defined baseline characteristics of mental health diagnoses in our population.

Methods: We examined SLC VAMC electronic medical records between January 1, 2014 and October 1, 2015 for encounters using the ICD-9 code for Gender Identity Disorder in Adolescents or Adults. Charts with the code were then examined for ICD-9 codes for tobacco use disorder, posttraumatic stress disorder (PTSD), depression, and anxiety-related conditions. Other ICD-9 codes for mental health diagnoses and suicide attempt/ideation requiring crisis contact were noted.

Results: Thirty-nine patients were found with the GID ICD-9 code. Of those, 31 (79.5%) identified as male-to-female, 8 (20.1%) female-to-male; 19 (48.7%) had a history of suicide attempt/ideation. PTSD was the most commonly identified mental health diagnosis (18/39 (46.1%)), depression second (16 (41.0%)), followed by tobacco use disorder (13 (33.3%)), other (substance abuse, bipolar disorder, schizotypal disorder) (9 (23.1%)), and anxiety (6 (15.4%)). When further examined, four (10.2%) patients were identified as having no mental health co-morbidities, 17 (42.6%) carried one mental health diagnosis and 18 (46.2%) carried 2 or more diagnoses (maximum number documented in one patient: 4 (10.2%)).

Conclusions: Of patients with GID at the SLC VAMC, 90% have mental health co-morbidities, nearly 50% having two or more diagnoses. Nearly 50% required intervention for suicide attempt/ideation. We have formed the multi-disciplinary Gender Dysphoria Team in an effort to reduce the impact of these diagnoses on the management of GID, however, further research into the optimization of management for this population is still needed.


Nothing to Disclose: MPG, BH, MB, KA, NA, TA, MG, BG, JH, CH, CK, RL, JM, AM, AT

OR10-5 24524 5.0000 A Prevalence of Co-Morbid Mental Health Diagnoses in Transgender Military Veterans at the Salt Lake City Veterans Affairs Medical Center.   2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Reproductive Endocrinology Friday, April 1st 1:15:00 PM OR10 7719 11:45:00 AM Recent Developments in Disorders of Sex Development (DSD) and Transgender Medicine Oral

Chantal M Wiepjes*, Mariska C Vlot, Maartje Klaver, Renate T de Jongh, Paul Lips, Annemieke C Heijboer and Martin den Heijer
VU University Medical Center, Amsterdam, Netherlands


Background: Estrogen has positive effects on bone mineral density (BMD), in particular in trabecular bone through inhibition of bone resorption. Testosterone increases bone size, but the effect on BMD is less clear. Therefore, cross-sex hormonal treatment (CSHT) in transgender patients can affect BMD.

Objectives: To investigate effects of CSHT on BMD during the first year of treatment in male-to-female (MtFs) and female-to-male transgender patients (FtMs). 

Methods: This study is a prospective observational study and part of ENIGI (European Network for Investigation of Gender Incongruence). 205 adult patients who completed one year of CSHT were included. In 107 FtMs and 98 MtFs a dual-energy X-ray absorptiometry was performed to measure lumbar spine BMD before and after a year CSHT. FtMs received intramuscular testosterone undecanoate (1000mg/12 weeks), testosterone gel (50mg/day) or testosterone esters intramuscular (250mg/2 weeks). MtFs were treated with oral estradiol valerate (2-4mg/day) or an estradiol patch (200ug/week). Most MtFs received cyproteronacetate (50mg/day) simultaneously.

Results: At baseline the mean lumbar spine BMD of FtMs was 1.02 g/cm2 (SD±0.13) and after one year 1.03 g/cm2 (SD±0.13), reflecting a mean increase of 1.12% (95%CI 0.32 – 1.92%). In MtFs at baseline and after one year, the lumbar spine BMD was 0.98 g/cm2 (SD±0.14) and 1.01 g/cm2(SD±0.14), respectively, reflecting a mean increase of 3.71% (95%CI 2.82 - 4.60%).

Conclusion: After one year CSHT the spine BMD increased in both groups, but more in MtFs. This confirms the role of estrogen on bone in biological males.


Nothing to Disclose: CMW, MCV, MK, RTD, PL, ACH, MD

OR10-6 26613 6.0000 A Effect of One Year Cross-Sex Hormonal Treatment on Bone Mineral Density of the Lumbar Spine in Transgender Patients 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Reproductive Endocrinology Friday, April 1st 1:15:00 PM OR10 7719 11:45:00 AM Recent Developments in Disorders of Sex Development (DSD) and Transgender Medicine Oral

Somik Chatterjee*1, Hongshan Yin2, Ji M Kim1, Zhe Fang3, David L Nelson3 and Ke Ma1
1Houston Methodist Research Institute, 2Third Affiliated Hospital, Hebei Medical University, 3Baylor College of Medicine


The circadian clock is intimately linked with metabolic homeostasis, and disruption of this mechanism leads to metabolic disorders including insulin resistance and obesity. In skeletal muscle, glucose and fatty acid oxidation is adapted to diurnal substrate availability, and we postulate that this metabolic switch may require the Bmal1-controlled intrinsic muscle clock coordination with the feeding-fasting cycle. By generating a mouse model with MCK-Cre transgene-mediated mature myocyte-specific ablation of the essential clock activator, Bmal1, here we demonstrate that the muscle-intrinsic clock is required for metabolic fuel utilization, and loss of this mechanism impacts metabolic substrate partitioning in muscle and liver. Bmal1 protein is strongly induced by feeding in slow muscle fibers, and its loss impairs the feeding-induced metabolic switching from fatty acids to glucose utilization. Consequently, muscle Bmal1-deficient mice display markedly reduced glucose metabolism but augmented fatty acid oxidation, without affecting mitochondrial function. Importantly, the increase in fat oxidation in skeletal muscle profoundly impacts whole-body metabolic homeostasis, resulting in significantly lower circulating lipids with suppression of hepatic lipid accumulation under fasting condition or high fat-diet challenge. In contrast, glucose level in muscle Bmal1-deficient mice was elevated accompanied by increased glycogen deposition in the liver. These metabolic actions of Bmal1 in skeletal muscle also resulted in resistance to high fat diet-induced obesity. Further mechanistic investigations reveal that Bmal1 coordinates the transcriptional regulation of key enzymes involved in fatty acid and glucose metabolism in accordance to the feeding-fasting signals in muscle. Taken together, our study demonstrates a critical role of the muscle clock in orchestrating the feeding and fasting-induced metabolic fuel flux between muscle and liver with significant impact on systemic metabolic homeostasis. Dysregulation of this temporal mechanism in coordinating inter-tissue metabolic cross-talk could be an under-appreciated component of the wide-spread metabolic abnormalities encountered with circadian misalignment in our modern lifestyle.


Nothing to Disclose: SC, HY, JMK, ZF, DLN, KM

OR06-1 25485 1.0000 A The Skeletal Muscle Circadian Clock Is Required for Metabolic Fuel Oxidation and Partition 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Nuclear Receptors and Steroid Hormone Action Friday, April 1st 1:15:00 PM OR06 7732 11:45:00 AM Nuclear Receptors and Co-Regulators in Health and Disease Oral

Tomoshige Kino*
Sidra Medical and Research Center, Doha, Qatar


Pediatric acute lymphoblastic leukemia (ALL) is a common and life-threatening malignancy in childhood. Glucocorticoids are among the most important therapeutic compounds for this leukemia, and responsiveness of leukemic cells to glucocorticoids is a valuable prognostic factor. Actions of glucocorticoids are mediated by the glucocorticoid receptor (GR), a member of the steroid/nuclear hormone receptor superfamily. The v-akt murine thymoma viral oncogene homolog 1p (AKT1) or the protein kinase B, a serine/threonine kinase that regulates cell proliferation, survival, migration and angiogenesis, is frequently activated in pediatric ALL, through various mutations/deletions in its upstream or nearby signaling molecules as well as in its own sequence. AKT1 is one of the causative agents for the insensitivity of leukemic cells to glucocorticoids, by phosphorylating a serine residue at position 134 (S134) located in the N-terminal domain (NTD) of human GR and by segregating phosphorylated GR from the nucleus in cooperation with 14-3-3. To further examine the effect of AKT1 and 14-3-3 on GR-induced transcriptional activity, we employed transient transfection-based reporter assays in GR-deficient human colon cancer HCT116 cells. We found that AKT1 had 2 independent actions on GR transcriptional activity, one is suppressive and dependent on S134, while the other is enhancing and independent to S134. 14-3-3 also had 2 effects on GR: one is dependent on S134 and suppressive to GR transcriptional activity only in the presence of active AKT1, and the other is independent to S134 and enhances the activity cooperating with AKT1. Using a GR mutant with large deletion in NTD and a GR chimera expressing its ligand-binding domain (LBD) fused with the GAL4 DNA-binding domain, we found that GR LBD supported the S134-independent enhancing activity of AKT1 and 14-3-3. Consistent with these results, 14-3-3 physically interacted with GR at 2 portions in co-immunoprecipitation assays: one is the S134 phosphorylated by AKT1 and the other is GR LBD. In the time course experiments examining the transcriptional activity of wild type GR or its S134A mutant coupled with their nuclear translocation profiles, we confirmed that AKT1 had dual actions on GR, the suppressive activity through phosphorylation of S134 and subsequent inhibition of the GR nuclear translocation, and the enhancing activity through direct modulation of GR transcriptional activity in the nucleus. These results suggest that specific inactivation of the AKT1 activity to phosphorylate GR at S134 is beneficial for the patients with glucocorticoid-resistant pediatric ALL, as it blocks the negative effect of AKT1 on GR that underlies insensitivity of leukemic cells to glucocorticoids, while it preserves/potentiates the beneficial, enhancing effect on the transcriptional activity of this receptor.


Nothing to Disclose: TK

OR06-2 25557 2.0000 A AKT1 Has Dual Actions on the Transcriptional Activity of the Glucocorticoid Receptor: Implications to Glucocorticoid Resistance Observed in Pediatric Acute Lymphoblastic Leukemia and Its Treatment 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Nuclear Receptors and Steroid Hormone Action Friday, April 1st 1:15:00 PM OR06 7732 11:45:00 AM Nuclear Receptors and Co-Regulators in Health and Disease Oral

Jaqueline de Carvalho Rinaldi*1, Wen Yang Hu1, Shyama Majumdar1, Dan Ping Hu1, Lishi Xie1, Timothy Gauntner1, Susan Kasper2, Luis Antonio Justulin3 and Gail S. Prins4
1University of Illinois at Chicago, Chicago, IL, 2University of Cincinnati, Cincinnati, OH, 3Universidade Estadual Paulista, Botucatu, Brazil, 4University of Illinois-Chicago, Chicago, IL


We previously demonstrated that estrogen regulates human prostate stem/progenitor cell amplification by directly targeting estrogen receptors (ERs); ERα stimulates whereas ERβ suppresses stem cell self-renewal. In addition to ERα and ERβ, we find that human prostate stem/progenitor cells express robust level of IGF-1R. Since ER actions can be modified by IGF-1R through ligand-independent ER phosphorylation, we herein sought to characterize potential cross-talk between estrogen and IGF-1 signaling pathways in regulating human prostate stem/progenitor cell amplification. Human prostate stem/progenitor cells were isolated from normal primary prostate epithelial cells (PrEC) using 3-D prostasphere (PS) culture. Two human prostate stem cell lines benign WPE-stem cells over-expressing ERα and HuSLC prostate cancer stem cells expressing ERβ but not ERα were used to explore the specific receptor subtype in estrogen action. Similar to estradiol-17β (E2), 5 nM IGF-1 treatment increased the number of PS as well as long-term BrdU-retaining prostate stem cells. Conversely, knockdown of IGF-1R by siRNA decreased both parameters and consistently increased PS ERβ expression. Together these findings suggest that IGF-1R activation may drive prostate stem cell amplification through suppression of ERβ. Further studies revealed that E2 (10 nM) exposure induced IGF-1R phosphorylation while IGF-1R knockdown inhibited the non-genomic E2-induced pAkt and pERK confirming the cross-talk between these two signaling pathways. Use of WPE and HuSLC cell lines revealed that E2 (10 nM) treatment induced IGF-1R phosphorylation through rapid signaling mediated by ERα but not ERβ. IGF-1R knockdown decreased PHLDA1, a known IGF-1 target gene, inhibited E2-induced ERα phosphorylation, suggesting a positive interaction between IGF-1R and ERα. In summary, the present results document robust crosstalk between estrogen and IGF-1 signaling which together regulate their downstream signal molecules including pAKT/pERK and PHLDA1. We propose that these pathways coordinately modulate prostate stem and progenitor cell numbers to effectively maintain glandular homeostasis.

Supported by FAPESP grant#2014/10965-6 and NIH/NCI award R01 CA172220.


Nothing to Disclose: JDCR, WYH, SM, DPH, LX, TG, SK, LAJ, GSP

OR06-3 26578 3.0000 A Cross-Talk Between Estrogen Receptors and Insulin-like Growth Factor Type-1 Receptor Modulates Human Prostate Stem/Progenitor Cell Amplification 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Nuclear Receptors and Steroid Hormone Action Friday, April 1st 1:15:00 PM OR06 7732 11:45:00 AM Nuclear Receptors and Co-Regulators in Health and Disease Oral

Imran Hussain*1, Chien-Cheng Chen1, Alina Peraza Montalbano1, Phuong Truong1 and Carole R Mendelson2
1UT Southwestern Medical Center, Dallas, TX, 2UT Southwestern Medical Center


Progesterone (P4), acting through its receptor (PR), plays a central role in the maintenance of pregnancy by suppression of proinflammatory (e.g. COX-2, IL-8) and contractile (e.g. CX43, OXTR) genes in the myometrium (1). P4/PR exerts these effects, in part, by tethering to nuclear factor-κB (NF-κB) bound to the promoters of these genes, resulting in a decline in NF-κB transcriptional activity (2). Recently, we found that the PR DNA-binding domain (DBD) plays a crucial role in P4-mediated suppression of endogenous proinflammatory genes; however, this inhibitory effect of P4/PR was not mediated by direct DNA binding. We further observed that P4/PR transrepression activity occurred at the level of transcription initiation and was mediated by decreased recruitment of NF-κB p65 and RNA Pol II to the COX-2 and IL-8 promoter regions. Thus, we postulated that nuclear proteins interacting with the PR DBD may play an important role in P4/PR mediated transrepressive activity. Using immortalized human myometrial cells (hTERT-HM) stably expressing wild-type PR-B (PR-BWT) or PR-B containing a mutation in the DBD (PRmDBD), we identified two transcriptional repressors, CtBP1 and GATAD2B, that interacted strongly with PR-BWT, but poorly with PR-BmDBD. P4 treatment of PRWT hTERT-HM cells, caused enhanced recruitment of endogenous GATAD2B and CtBP1 to NF-κB-binding regions of the COX-2 and IL-8 promoters. Further, siRNA knockdown of endogenous GATAD2B or CtBP1 significantly reduced P4/PR-BWT transrepression of COX-2 and IL-8. Using RT-qPCR, we observed that GATAD2B and CtBP1 mRNA levels were significantly decreased in myometrial biopsies of pregnant women in-labor, as compared to those from women not-in-labor, at term. To gain further insight into expression and function of GATAD2B and CtBP1 in the regulation of contractile gene expression during pregnancy and parturition, we analyzed myometrial tissues from timed pregnant mice. We observed that increased expression of Oxtr and Cx43 in mouse myometrium near term was associated with a marked decline in Gatad2b and Ctbp1 mRNA and protein. Using ChIP-qPCR, we found that increased expression of Oxtr and Cx43 in myometrium near term occurred in concert with a marked decline in binding of endogenous Gatad2b and Ctbp1 to the NF-κB-binding regions of the Oxtr and Cx43 promoters. This decline in corepressor recruitment was associated with a pronounced decrease in the repressive histone mark, H3K27me3. Together, these findings suggest that Gatad2b and Ctbp1 serve as novel mediators of P4/PR suppression of myometrial proinflammatory and contractile gene expression. The decreased expression and DNA-binding of these corepressors near term contribute to the decline in PR function and result in the induction of myometrial proinflammatory and contractile genes leading to labor.


Nothing to Disclose: IH, CCC, APM, PT, CRM

OR06-4 26681 4.0000 A C-Terminal Binding Protein 1 (CtBP1) and GATAD2B Serve As Novel Mediators of Progesterone/PR Suppression of Proinflammatory and Contractile Genes in the Pregnant Myometrium 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Nuclear Receptors and Steroid Hormone Action Friday, April 1st 1:15:00 PM OR06 7732 11:45:00 AM Nuclear Receptors and Co-Regulators in Health and Disease Oral

Anna S Garza1 and Raj Kumar*2
1University of Texas Medical Branch, Galveston, TX, 2The Commonwealth Medical College, Scranton, PA


Like other members of the steroid hormone receptor family, phosphorylation of the glucocorticoid receptor (GR) is a critical step in the receptor activation and biological activity.  In the human GR, the major functionally important known phosphorylation sites are serine S203, S211, and S226. All of these sites are located within the intrinsically disordered (ID) AF1 domain. We and others have shown that p38-mediated phosphorylation of S211 enhances the ability of the receptor to regulate transcription and apoptosis. Later, we reported that phosphorylation of S211 induces a functionally active conformation in the ID AF1 domain such that AF1’s interaction with specific coregulatory proteins and subsequent transcriptional activities are significantly enhanced. Based on these findings, we proposed that phosphorylation-induced disorder-order conformational transition may be a potential mechanism through which site-specific phosphorylation regulates GR functions. However the exact role of the other two phosphorylation sites S203 and S226 remains unclear. It has earlier been reported that relative level of site-specific phosphorylation of GR is an important determinant of receptor activity in a gene-specific manner. Therefore, in this study we examined the role of S203 and S226 phosphorylation sites on the structure and functions of AF1 and compared these effects with S211. Our limited tryptic digestion experiments showed that in vitro phosphorylated recombinant AF1 and AF1S226A, and un-phosphorylated AF1S211E and AF1S226E resist proteolysis by trypsin when compared to phosphorylated AF1S211A and un-phosphorylated AF1 suggesting that S211 may be the main phosphorylation site that is responsible for conformational changes in AF1. Further, CV-1 cells constitutively expressing AF1 in a two domain GR fragment containing entire N-terminal and DNA-binding domains (GR500), or mutants were co-transfected with DNA of the pGRE-SEAP plasmid alone or plus DNA for TBP. Fluorescence resonance energy transfer (FRET) to examine protein-protein interactions between GR500 and TBP and SEAP assay to determine promoter-reporter activity were carried out. We found that though GR500-S203A or GR500-S226A mutants show increased FRET for TBP, yet this TBP induced FRET increase of these mutants do not reach that of GR500 and TBP. On the other hand, the GR500 containing triple mutants (S203, S211A, S226A) lost most of TBP interaction. Similar results were obtained for AF1-mediated GR promoter-reporter activity. Together, these results demonstrate the role of different phosphorylation sites in the regulation of AF1-mediated GR actions.


Nothing to Disclose: ASG, RK

OR06-5 27070 5.0000 A Role of Site-Specific Phosphorylation in the Action of Glucocorticoid Receptor 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Nuclear Receptors and Steroid Hormone Action Friday, April 1st 1:15:00 PM OR06 7732 11:45:00 AM Nuclear Receptors and Co-Regulators in Health and Disease Oral

Raymond E Soccio*, Hee-Woong Lim, Eric R Chen, Joanna R Dispirito, Satyajit R Rajapurkar, Erika R Briggs, Shannon E Mullican, David J Steger, Kyoung Jae Won and Mitchell A Lazar
University of Pennsylvania, Philadelphia, PA


The nuclear receptor PPARγ is the master regulator of fat cell development and implicated in type 2 diabetes as the target of insulin-sensitizing thiazolidinedione drugs.  High fat diet (HFD)-induced obesity is a common mouse model of metabolic syndrome, in which white adipose tissue (WAT) becomes infiltrated with inflammatory cells, notably macrophages.  PPARγ is also expressed in macrophages, where we have previously reported a set of unique genomic binding sites not present in cultured 3T3-L1 adipocytes (1).  Here we used chromatin immunoprecipitation followed by deep sequencing (ChIP-Seq) to describe genome-wide binding regions (cistromes) for PPARγ in WAT from male C57BL/6 mice fed a control low fat diet (10% calories from fat) versus HFD (60% calories from fat) for 4 or 12 weeks, which resulted in marked remodeling of the PPARγ cistrome in epididymal WAT (eWAT).  Remarkably, those PPARγ sites gained in eWAT upon HFD were highly enriched for macrophage-selective binding sites.  These were lost in eWAT of mice with macrophage-specific PPARγ deletion, confirming their macrophage-specificity and indicating detectable PPARγ genomic binding events in WAT-resident macrophages.  These changes in PPARγ occupancy at regulatory elements correlated with nearby mRNA expression, as genes involved in macrophage and inflammatory pathways are induced in eWAT from HFD-fed mice.  Conversely, loss of PPARγ binding in eWAT upon HFD was observed near adipocyte-selective binding sites, and the HFD-repressed genes are involved in adipocyte and metabolic pathways.  Similar dynamic patterns were observed at the epigenome, notably in histone acetylation.  In contrast, 129S1/SvIm strain mice, which are resistant to diet-induced obesity, showed little change in PPARγ occupancy or gene expression in eWAT upon HFD-feeding.  This integrative genomic analysis revealed that HFD-regulated changes in adipose tissue PPARγ cistromes begin early, prior to marked weight gain, inflammation, and insulin resistance, differentially impacting the function of adipocytes and adipose tissue macrophages.


Disclosure: MAL: Board Member, Eli Lilly & Company, Board Member, Pfizer, Inc., Ad Hoc Consultant, Jansen Pharmaceuticals, Ad Hoc Consultant, KDAC Therapeutics, Inc, Ad Hoc Consultant, Synageva BioPharma Comp.. Nothing to Disclose: RES, HWL, ERC, JRD, SRR, ERB, SEM, DJS, KJW

OR06-6 27520 6.0000 A High Fat Diet-Induced Remodeling of Pparγ Binding Sites in Mouse White Adipose Tissue 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Nuclear Receptors and Steroid Hormone Action Friday, April 1st 1:15:00 PM OR06 7732 11:45:00 AM Nuclear Receptors and Co-Regulators in Health and Disease Oral

Mandy Geserick1, Mandy Vogel2, Ruth Gausche1, Roland Pfäffle2, Wieland Kiess2 and Antje Körner*2
1University of Leipzig, Dept. of Women´s & Child Health, 2University of Leipzig, Dept. of Women´s & Child Health, Leipzig, Germany


We aimed to assess the predictive value of BMI at earlier ages for the risk of developing obesity in adolescence and to identify vulnerable age periods that may be crucial for subsequent overweight/obesity.

We retrieved anthropometric data of 33.537 subjects from the CrescNet® data base and included those with measurements of at least two time points.

Overweight and obesity were present in 10% and 9% of the adolescents (15-18 years), respectively. Changes from normal weight to overweight/obesity in childhood were considered and probability of being overweight/obese in adolescence given a certain BMI-SDS in childhood was calculated.

Retrospectively seen, 53% of the children, who were obese in adolescence were normal weight until the age of 4 years. By the age of 5 years, however, half of them were overweight/obese. Prospectively, 60% of children who are overweight already at the age of 4 years remained overweight/obese in adolescence. The probability of being overweight/obese in adolescence in those who are already obese at the age of 4 was 79%. In a subgroup of 26.344 children we considered the influence of BMI acceleration in different age periods. Dynamically, children in the age of 2 to 5 years, who had a rise in BMI-SDS by 0.2-2 during 1 year had a higher risk of being overweight/obese in adolescence than older children with the same BMI acceleration.

In conclusion, overweight/obesity in adolescence is already determined in early childhood. Periods of increased BMI acceleration in early childhood should be recognised for an optimized strategy in obesity prevention.


Nothing to Disclose: MG, MV, RG, RP, WK, AK

OR07-1 27395 1.0000 A Tracking of BMI from Infancy to Adolescence and Predictive Value for the Development of Obesity 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Obesity, Adipocyte Biology and Appetite Friday, April 1st 1:15:00 PM OR07 7739 11:45:00 AM Predictors of Weight Gain and Disease Oral

Joanna Huang1, Sarah Buchs1, Maral DerSarkissian*2, Rachel Bhak3, Francis Vekeman4, Rahul Ganguly1 and Mei Sheng Duh3
1Novo Nordisk, Inc., Plainsboro, NJ, 2Analysis Group, Inc., Los Angeles, CA, 3Analysis Group, Inc., Boston, MA, 4Groupe d’Analyse, Ltée, Montreal, QC, Canada


Background: Achieving and maintaining weight loss (WL) has proven to be difficult as many patients regain weight after an initial WL.  To understand patterns of weight change among patients with obesity, we conducted a retrospective longitudinal study of patients with obesity using the General Electric Centricity electronic medical record database.  Identifying patterns of weight change is critical for tailoring weight management strategies to the needs of targeted patient groups.   

Methods: The study sample included patients age ≥18 years old with BMI ≥ 30 kg/m2 (first observed BMI measurement was defined as index BMI) who had no medical conditions associated with unintentional WL and had ≥4 BMI measurements per year for ≥5 years.  Patients were categorized into 4 weight groups based on their amount of weight change during a WL period of two quarters following the index BMI: stable weight= within <5% of index BMI; modest WL= ≥5 to <10% of index BMI lost; moderate WL= ≥10 to <15% of index BMI lost; and high WL= ≥15% of index BMI lost.  Patterns of weight change were also assessed following the WL period for 8 consecutive quarters (i.e., the weight maintenance [WM] period).  Subgroup analyses were conducted among patients with class II obesity (index BMI≥35), pre-diabetes, and type 2 diabetes (based on diagnoses and lab values).

Results: Of 194,490 patients, 177,743 patients were included in the main analysis as they were classified in one of the 4 weight groups of interest during the WL period: 151,236 (85.1%) patients were in the stable weight group, 16,559 (9.3%) were in the modest WL group, 4,017 (2.3%) were in the moderate WL group, and 5,931 (3.3%) were in the high WL group.  Though 7.2% of the modest WL group continued to lose weight in the first quarter of the WM period, this dropped to 2.0% by the eighth quarter; in the moderate WL group these proportions were 13.3% and 4.1%; and 18.8% and 11.1% for the high WL group.  A decreasing trend in the overall proportion of patients who maintained their weight throughout the WM period was also observed.  In the modest, moderate, and high WL groups 40.0%, 35.9%, and 18.6%, of patients, respectively, regained ≥50% of lost weight during the WM period.  Cyclers were defined as patients who did not consistently lose, maintain, or gain weight in each quarter of the WM period.  The high WL group had the lowest proportion of cyclers with 58.3%, while 71.5% of the modest WL group and 74.1% of the moderate WL group were cyclers.  This trend was similar in the subgroups analyzed, though among patients with class II obesity those with high initial WL were more likely to continue to lose weight during the WM period compared to the overall sample and other subgroups.      

Conclusion: Weight cycling and regain was commonly observed among patients.  Patients who lost more weight during the WL period (overall and in subgroups) were more likely to keep the weight off and continue losing weight.


Disclosure: JH: Employee, Novo Nordisk, Employee, Novo Nordisk. SB: Employee, Novo Nordisk. MD: Researcher, Novo Nordisk. RB: Researcher, Novo Nordisk. FV: Researcher, Novo Nordisk. RG: Employee, Novo Nordisk, Employee, Novo Nordisk. MSD: Researcher, Novo Nordisk.

OR07-2 24110 2.0000 A Patterns of Weight Loss, Maintenance, and Gain in Patients with Obesity 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Obesity, Adipocyte Biology and Appetite Friday, April 1st 1:15:00 PM OR07 7739 11:45:00 AM Predictors of Weight Gain and Disease Oral

Rebecca J Gordon*, Sunil K. Panigrahi, Kana Meece, Deniz Atalayer, Richard M. Smiley and Sharon L. Wardlaw
Columbia University College of Physicians & Surgeons, New York, NY


The melanocortin neuronal system, consisting of the proopiomelanocortin (POMC)-derived MSH peptides and the MSH antagonist, agouti-related protein (AgRP), plays a key role in regulating energy balance. Endogenous opioid regulation of this system at multiple levels has been demonstrated in animals. POMC neurons express μ-opioid receptors which can function as inhibitory autoreceptors in response to release of the POMC-derived peptide, ß-endorphin (ß-EP). The opioid antagonist, naltrexone (NTX), has well established stimulatory effects on POMC neurons and can decrease food intake in rodents. NTX is also part of a new FDA approved weight loss combination drug together with bupropion. We have previously shown that NTX acutely stimulates POMC peptide release (MSH and ß-EP) in the rodent, accompanied by a fall in hypothalamic peptide content by 2 days and a subsequent stimulation of POMC mRNA levels after 7 days. Little is known about the effects of NTX on brain POMC in humans. We have therefore studied the effects of NTX on the brain melanocortin system in human subjects as assessed by cerebrospinal fluid (CSF) neuropeptide concentrations. Eleven healthy subjects were studied in a double-blind crossover study with NTX (50 mg po daily at 9 pm) or placebo for 2 days (n=5) or 7 days (n=6). CSF was collected by lumbar puncture in the morning after a 12h overnight fast and a blood sample was also obtained. CSF POMC was measured by in-house ELISA that detects the POMC prohormone; ß-EP was measured by in-house ELISA that is specific for ß-EP and does not crossreact with POMC. AgRP was measured by ELISA (R&D Systems) in CSF and plasma as both may reflect brain AgRP activity. There was no significant effect of NTX on CSF POMC levels. However after NTX, CSF ß-EP increased to 172% of the placebo level (p=0.002); the increase was significant in both the 2 and 7 day administration groups. The ß-EP to POMC ratio also increased after NTX to 175% of placebo. There was no significant change in CSF or plasma AgRP but plasma AgRP tended to increase after NTX (p=0.13). Thus, opioid antagonism with NTX stimulates POMC peptide release into CSF in humans, consistent with effects on brain POMC peptide release in rodents. No effect on POMC prohormone levels was seen after 1 week. The increase in the CSF ß-EP to POMC ratio is consistent with selective release of the processed peptides or an effect on POMC processing. Furthermore, a potential stimulatory effect of NTX on AgRP could serve to mitigate the stimulatory effects on POMC peptide release with respect to decreasing food intake. It remains to be determined if biomarkers in CSF and plasma could be used to predict responses to obesity pharmacotherapy targeting the melanocortin system.


Nothing to Disclose: RJG, SKP, KM, DA, RMS, SLW

OR07-3 24462 3.0000 A Effects of Opioid Antagonism on CSF Proopiomelanocortin and ß-Endorphin Levels in Human Subjects 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Obesity, Adipocyte Biology and Appetite Friday, April 1st 1:15:00 PM OR07 7739 11:45:00 AM Predictors of Weight Gain and Disease Oral

Shana E. McCormack*1, Sarah E. Henrickson2, Zhe Zhang2, E. John. Wherry3, Thomas Wadden4 and Robert I. Berkowitz2
1The Children's Hospital of Philadelphia, Philadelphia, PA, 2Children's Hospital of Philadelphia, Philadelphia, PA, 3University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, 4University of Pennsylvania, Philadelphia, PA


Background: Currently, there are no well-validated strategies to individualize recommendations for management of pediatric obesity, despite the considerable variability in treatment responses. 

Objective: To identify novel biomarkers in obese adolescents, assessed at the time of study enrollment, that predict successful response to a 4-month lifestyle modification intervention using non-targeted proteomics and metabolomics.

Methods: The primary outcomes of this behavioral weight loss trial are published.(1) Outcome measures included 4- and 12-month changes in body mass index (BMI), proteomics (1126 proteins, SOMAScan, SomaLogic, Inc), metabolomics (417 metabolites, Metabolon, Inc). Normalized protein and metabolite levels were compared between weight loss response groups using rank product (RP) testing (2) to obtain p values and false discovery rates (FDRs) through a permutation procedure. Proteins/metabolites were also mapped to curated sets for pathway analysis.

Results: Participants (n=40) were 74% female, 38% African-American, 14.9 years old (±1.2 SD), with baseline BMI Z-score of 2.31 (±0.29 SD). On average, BMI decreased by 7.2±1.3% after 4 months, and decreased by 5.5±2.2% after 12 months. Subjects were divided into 4 groups based on their weight loss responses: “Group 1” (n=13), strong 4- and 12-month responses, “Group 2” (n=10), strong 4-month response, but some weight regain by 12 months, “Group 3” (n=15), modest but consistent response over 4 and 12 months, and “Group 4” (n=9), poor response at both 4 and 12 months. We prioritized comparisons between “Group 1” (treatment-responsive) and “Group 4” (treatment-resistant) participants.

The most apparent group-specific differences were levels of immunomodulatory and neutrophil associated proteins that were higher in treatment-resistant participants (p<0.001, FDR <1%, for both). In addition, fibroblast growth factor 19 (FGF19), an insulin-independent modulator of hepatic protein and glycogen synthesis, was also significantly higher in treatment-resistant subjects (p=0.0001, FDR=1%). Finally, levels of proteins related to metabolism of amino acids (Reactome R-HSA-7121) were different between treatment-resistant versus treatment-responsive obesity (p=0.0016, FDR=3%), and levels multiple individual protein and amino acid derivatives were also nominally different, e.g., gamma-glutamylmethionine (higher in treatment-resistant group, p=0.0048, FDR=25%).

Conclusions: We speculate that increased activity of immunomodulatory and neutrophil associated pathways is related to treatment-resistant obesity in adolescents. In the future, leveraging biomarker profiles may facilitate individualized, rational treatment formulations for pediatric obesity. Investigation of the above signals in a separate validation cohort is ongoing.


Disclosure: TW: Advisory Group Member, Nutrisystem, Advisory Group Member, Orexigen Pharmaceuticals, Advisory Group Member, Novo Nordisk, Advisory Group Member, Weight Watchers. Nothing to Disclose: SEM, SEH, ZZ, EJW, RIB

OR07-4 25689 4.0000 A Integrated Proteomic and Metabolomic Profiles of Successful Weight Loss in Obese Adolescents 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Obesity, Adipocyte Biology and Appetite Friday, April 1st 1:15:00 PM OR07 7739 11:45:00 AM Predictors of Weight Gain and Disease Oral

Allison R. Smego*1, Jessica G Woo1, Jillian Klein2, Danesh Bansal3, Christopher F. Bolling4, Stephen R. Daniels5 and Nancy Crimmins1
1Cincinnati Children's Hospital Medical Center, Cincinnati, OH, 2Cincinnati Children's Hospital Medical Cener, Cincinnati, OH, 3Northeast Ohio Medical University, 4Pediatric Associates, Crestview Hills, KY, 5Children’s Hospital of Colorado, Denver, CO


Background: Young children with severe obesity have high lifetime risk for persistent obesity and metabolic disease. It is unknown when young children with severe obesity develop abnormal weight gain patterns thus limiting early identification and intervention. Our objective was to characterize growth patterns of children who become severely obese (BMI>99%ile) by age 6 to identify whether these children experience a critical period of deviation from normal growth, and to characterize that growth relative to normally-developing children.  

 Methods: Two lean cohorts (BMI 5th to ≤75%ile; one from low-income primary care clinic, one from longitudinal epidemiologic study) and two severely obese cohorts (BMI≥99%ile; one from low-income primary care clinic, one from obesity referral clinic) were selected based on BMI between ages 2-6.  Growth data were abstracted from birth through age 6 for each. In addition, sex, race, and health insurance type were collected. Repeated measures mixed modeling and logistic regression were used to distinguish growth characteristics of the lean and obese cohorts.

 Results: 783 lean (647 low-income, 136 population-based) and 480 severely obese (365 low income, 115 referral clinic) participants were included. Sex distribution of patients was similar among all groups. Both the low-income populations were predominantly African-American at 79% and 62%, respectively, while the population-based and referral clinic had a significantly lower proportion of African-Americans, at 24% and 23%, respectively. Clinical obesity onset occurred at median 2.00 [IQR: 1.24, 3.11] years of age in the low income obese, and at 1.36 [0.61, 3.02] years in the referral obese group. However, BMI differed significantly between severely obese and lean cohorts by 4 months of age (p<0.0001) while WHO weight-for-age percentile differed by 2 months (p<0.0001), prior to obesity onset. BMI high-specificity (95%) thresholds accurately differentiated severely obese from lean groups at 6, 12, and 18 months of age (51%, 52% and 95% sensitivity, respectively).

 Conclusions:  BMI trajectories in children who become severely obese by age 6 differ from children who remain normal weight as early as 4-6 months of age, approximately 12-18 months prior to the median onset of clinical obesity. Infant BMI at 6, 12, or 18 months above 85th percentile on WHO growth chart is highly specific for identifying children at risk for early onset severe obesity and should prompt close monitoring.


Nothing to Disclose: ARS, JGW, JK, DB, CFB, SRD, NC

OR07-5 24744 5.0000 A BMI Trajectory of Severely Obese Children Diverges from Normal-Weight Children during Infancy 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Obesity, Adipocyte Biology and Appetite Friday, April 1st 1:15:00 PM OR07 7739 11:45:00 AM Predictors of Weight Gain and Disease Oral

Amir H Sam1, Adam Buckley*2, Karim Meeran1, Paul Bech3, Maha Taysir Barakat2, Stephen R Bloom4, Nader Lessan2 and Kevin G. Murphy3
1Imperial College NHS Healthcare Trust, London, United Kingdom, 2Imperial College London Diabetes Centre, Abu Dhabi, United Arab Emirates, 3Imperial College London, London, United Kingdom, 4Imperial College London, United Kingdom


Fasting plasma pancreatic polypeptide (PP) predicts visceral adiposity and intrahepatocellular lipid concentration (1), Visceral adiposity is associated with vascular risk in type 2 diabetes mellitus (T2DM) and the metabolic syndrome.  We hypothesised that elevated fasting PP would be associated with microvascular complications of T2DM.  We enrolled 498 adult participants with normal glucose tolerance (NGT, n=160), impaired fasting glucose / impaired glucose tolerance (IFG/IGT, n = 92) or T2DM (n = 246).  Our endpoint was evidence of microvascular disease as indicated by diabetic retinopathy or persistent microalbuminuria.  

Using case matching to control for differences in age, sex, body mass index (BMI) and estimated glomerular filtration rate (eGFR), fasting PP was significantly higher in individuals with T2DM than either NGT (T2DM n = 105, PP median 35.0 pmol/L (interquartile range 21.7-62.2); NGT n = 105, PP 25.2 pmol/L (12.8-31.1); p < 0.0001) or IFG/IGT (T2DM n = 84, PP 51.6 pmol/L (21.6-62.8); IFG/IGT n = 84; PP 23.4 (15.8-46.7); p = 0.002), while PP did not differ significantly between individuals with IFG/IGT  and those with NGT.  This difference was particularly interesting given that the clinical distinction between T2DM and IFG/IGT is predicated on the greatly increased vascular disease risk in T2DM. 

Among individuals with T2DM and with adequate retinal screening within 1 year of the study date (n=236), we found that PP was significantly higher in those with background retinopathy (66.1 pmol/L (38.5-102.3), p < 0.01) or moderate non-proliferative or proliferative retinopathy (61.0 pmol/L (40.29-91.05) p < 0.05) than individuals without retinopathy (35.58 pmol/L (21.68-64.77)).   In common with other peptide hormones, fasting PP is increased in renal impairment due to reduced filtration and metabolism.  However, PP was significantly elevated in participants with T2DM who showed evidence of persistent microalbuminuria (50.3 pmol/L (28.4-75.0) cf. 35.0 pmol/L (21.7-65.0), p = 0.046) when those with eGFR < 60 mL/min/1.73m2 were excluded. We used a logistic regression model to further investigate the association of elevated PP with diabetic retinopathy.  Log-transformed PP remained a significant predictor of retinopathy when age, sex, duration of diabetes, HbA1c, systolic blood pressure, eGFR and log-transformed serum triglycerides were included as covariates (p = 0.039, OR =  3.672 per 10-fold increase in PP).  

Measuring visceral adiposity is expensive and difficult to do accurately in a clinical context.  We speculate that PP not only acts as a measure of visceral adiposity but may also reflect its contribution to vascular risk.  Given that retinopathy is associated with, and usually precedes, other microvascular and macrovascular complications of T2DM, the association we describe raises the possibility that fasting PP measurement may be of use in vascular risk stratification.


Nothing to Disclose: AHS, AB, KM, PB, MTB, SRB, NL, KGM

OR07-6 24226 6.0000 A Fasting Pancreatic Polypeptide Is a Potential Biomarker for Microvascular Disease in Type 2 Diabetes Mellitus 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Obesity, Adipocyte Biology and Appetite Friday, April 1st 1:15:00 PM OR07 7739 11:45:00 AM Predictors of Weight Gain and Disease Oral

Daniel Fiordelisio de Carvalho*1, Giselle Yuri Hayashi2, Mirela Costa de Miranda1, Helena Panteliou Lima Valassi3, Atecla Nunciata Lopes Alves4, Andresa De Santi Rodrigues5, Larissa Garcia Gomes6, Guiomar Madureira4, Berenice B Mendonca7 and Tania A Bachega1
1Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brazil, 2Laboratório da APAE SÃO PAULO, Brazil, Sao Paulo, 3Laboratório de Hormônios e Genética Molecular- LIM/42, Hospital das Clínicas, Faculdade de Medicina da Universidade de São Paulo, Sao Paulo, Brazil, 4Laboratório de Hormônios e Genética Molecular- LIM/42, Unidade de Adrenal, Disc. de Endocrinologia e Metabologia, Hospital das Clínicas, Faculdade de Medicina da Universidade de São Paulo, Sao Paulo, Brazil, 5Laboratório de Hormônios e Genética Molecular- LIM/42, Unidade de Adrenal, Disc. de Endocrinologia e Metabologia, Hospital das Clínicas, Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brazil, 6Laboratório de Hormônios e Genética Molecular- LIM/42, Disc. de Endocrinologia e Metabologia, Hospital das Clínicas, Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brazil, 7Unidade de Endocrinologia do Desenvolvimento, Laboratório de Hormônios e Genética Molecular LIM/42, Disciplina de Endocrinologia, Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo


Introduction: Congenital adrenal hyperplasia newborn screening (NBS) presents high capacity to detect the salt-wasters (SW); however, main concerns are the high false-positive results (FPR) rate, low positive predictive value (PPV) of neonatal (N) 17OHP levels and the heterogeneity of confirmatory test methods. Considering the recent CAH-NBS implementation in our country, our objectives are (1) to optimize the best cutoff levels for the N17OHP in the first test and (2) to evaluate the best serum confirmatory test.

Materials and methods: data of 473,983 newborns (NBs) were retrospectively evaluated. N17OHP was measured by IFMA (AutoDelfia) and cutoffs (99th and 99.8th percentiles) adjusted according to birthweight (BW1:<1500g; BW2:1500-2000g; BW3:2001-2500g; BW4: >2500g), and to age at sample collection (before or after 72hs of life). For confirmatory tests, serum 17OHP analyses were performed by RIA and LC-MS/MS and 21-deoxicortisol (21DF), Δ4 and cortisol levels by LC-MS/MS. Asymptomatic NBs with persistently increased serum 17OHP levels had entire CYP21A2 sequenced in peripheral DNA samples.

Results: the recall rate was 0.05% (n=221) using the P99th of N17OHP levels and decreased to 0.03% (n=149) using the P99.8th; additionally, PPV increased from 11% (P99th) to 17% (P99.8th). N17OHP cutoffs in samples collected before 72hs of life were significantly lower than those collected after. Since in our state since most samples are collected earlier, different cutoffs according to BW and age at sample collection were determined. Twenty-six NBs were diagnosed (22 SW, 4 SV, 12 males), confirmed by sequencing. N17OHP levels ranged from 53-494 ng/mL (serum equivalence) in SW form and from 36-53 ng/mL in SV form. Serum confirmatory tests were performed in 149 NBs. In affected NBs, serum 17OHP levels (LC-MS/MS) ranged from 56-668 ng/mL in SW and from 54-117 ng/mL in SV form. FPR persisted using 17OHP measurements in 70% of samples by RIA and 13% by LC-MS/MS. PPV of LC-MS/MS methodology was significantly higher than RIA (52 vs.27%). Serum 21DF and steroid ratios (17OHP/cortisol; 17OHP+Δ4/cortisol; 17OHP+21DF/cortisol) presented similar FPR and PPV values in comparison to 17OHP by LC-MS/MS. Serum 21DF and steroid ratio values overlapped between affected NBs and those with FPR. Among asymptomatic NBs with persistently increased serum 17OHP levels, molecular analysis identified 2 with NC form.

Conclusions: N17OHP levels adjusted to P99.8th and to sample collection time improve the CAH-NBS by reducing the FPR rate without missing the diagnosis of classical forms. Although serum 17OHP by RIA is widely used as confirmatory test in our country, the use of 17OHP by LC- MS/MS significantly reduced the FPR. The 21DF and steroid ratio measurements did not provide higher accuracy than serum 17OHP by LC-MS/MS. Molecular analysis could be restricted for asymptomatic NBs with persistently increased 17OHP levels.


Nothing to Disclose: DFDC, GYH, MCDM, HPLV, ANLA, ADSR, LGG, GM, BBM, TAB

OR08-1 26282 1.0000 A Newborn Screening for Congenital Adrenal Hyperplasia: Improving the Effectiveness of the Neonatal 17OH-Progesterone and Serum Confirmatory Tests 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Pediatric Endocrinology Friday, April 1st 1:15:00 PM OR08 7755 11:45:00 AM Pediatric Endocrinology Oral

Uta Neumann1, Erwin Lankes2, Franziska Bathelt-Tok2, Richard J Ross3, Heiko Krude4 and Oliver Blankenstein*1
1Charité Universitätsmedizin Berlin, Berlin, Germany, 2Charité Universitätsmedizin Berlin, 3University of Sheffield, United Kingdom, 4Charité - Universitätsmedizin Berlin Campus Virchow, Berlin, Germany


Background: Treatment of CAH in children is compromised by the pharmacokinetic of available hydrocortisone (HC) preparations resulting in un-physiological early morning rise of ACTH followed by elevated androgens. Usually CAH-therapy is monitored by blood sampling at a randomly time point during the day with therapy following a fixed dosing scheme (50% - 25% - 25%). We investigated individualized optimization of HC-treatment using alternative dosing-schemes and monitoring of 17OHP in saliva.

Treatment protocol: HC-treatment in children with CAH is applied 8 hourly  with a 1st dose (D1) in the morning (6-8h) , 2nd dose (D2) in the early afternoon (14-16h) and 3rd dose(D3) late at night (22-24h). Therapy monitoring used saliva-17-OHP profiles collected at home directly prior to the HC medication (5 - 10fold concentrations of age-specific reference range used as target range). In case of out of range salivary 17-OHP profiles the HC dose prior to the sampling point was lowered or increased. Patients are followed clinically (and by saliva profile) every 3 months, blood-sampling takes place once a year at the clinic appointment. In a sub-cohort of children participating in the pharmacokinetic part of the TAIN-treatment trial blood sampling was timed exactly prior the morning dose of HC.

Methods: We describe the dosing scheme after individualized adaptation by 17OHP saliva profiles of 31 children aged 1-11 years (females n=15, males n=16) in a longitudinal period of 2 years. Growth indicators, dosing and treatment adaptation frequency at the different medication times as well as ACTH and androgen levels in randomly or timed blood sampling were used as outcome parameters.  8 infants started saliva sampling within the study period.

Results: The resulting dosing scheme based on our “pre-dose-at-home-saliva-sampling” is different from published recommendations (40 – 20 – 40%).  Dose adaptation frequency was 59/248 visits (30% D1, 29% D2, 41% D3). Single dose adaptation occurred in 65%, two-dose adaptation in 20% and three-dose adaptation in 15%.   Mean total dose at T0 (10,77 mg/sqm/d) / T-12 month (10,88 mg/sqm/d) / T-24 month (11,03 mg/sqm/d)  was declining during the study period. Blood androgen levels were different between randomly and timed sampling. Height SDS and weight SDS remained stable in saliva controlled patients.

Conclusions: Dose adaptation by saliva sampling combined with 8hourly dosing scheme results in a rise of the late night dose and lowering the afternoon doses. Total HC doses are within the recommended ranges for HC replacement-doses in CAH-children. Regularly saliva profiling is easy to perform and leads to an individualized treatment based on a more frequent dose-adaptation. Saliva sampling was well tolerated and without stress for the children. Random and timed blood androgen levels are of limited value for therapy optimization.


Disclosure: RJR: Consultant, Diurnal. Nothing to Disclose: UN, EL, FB, HK, OB

OR08-2 26929 2.0000 A Optimization of Hydrocortisone Treatment in Children with Congenital Adrenal Hyperplasia (CAH) with Home 17OHP-Saliva Sampling: Feasibility and Effectiveness 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Pediatric Endocrinology Friday, April 1st 1:15:00 PM OR08 7755 11:45:00 AM Pediatric Endocrinology Oral

Tero Varimo*1, Matti Hero1, Kirsi Tuuli Vaaralahti2, Leo Dunkel3, Paivi Miettinen4 and Taneli Raivio5
1Helsinki University Hospital (HUH), Helsinki, Finland, 2University of Helsinki, Faculty of Medicine, Helsinki, Finland, 3Centre for Endocrinology, William Harvey Research Institute, London, United Kingdom, 4Helsinki University Hospital (HUH), espoo, Finland, 5University of Helsinki, Helsinki, Finland


BACKGROUND: Makoring ring finger protein 3 (MKRN3) gene restrains the hypothalamic-pituitary-gonadal (HPG) axis and thereby controls the onset of puberty (1). In girls, peripheral levels of MKRN3 decline prior to the onset of puberty (2), whereas in boys the changes in serum MKRN3 levels before and during puberty have not been reported.

PATIENTS AND METHODS:  This randomized controlled study included 30 peripubertal boys (age range 9.1-14.2 yrs) with idiopathic short stature (ISS) (3). Sixteen boys were treated with letrozole (2.5mg/d) for 2 yrs and 14 received placebo. Boys were followed up for 3 yrs, hormonal and MKRN3 levels were obtained with 6 mo intervals for 2 yrs, and analyzed using summary measures.

RESULTS: The boys showed an age-dependent decline in serum MKRN3 levels (mean regression coefficient -6.6±7.2 pg/mL per year, P < 0.001), with no difference between the two groups. Importantly, MKRN3 levels declined before Tanner genital stage 2, but not thereafter (-29.3±27.5 vs -5.6±15.1 pg/mL per year) (P < 0.05). During Tanner genital stage 1, the rate of MKRN3 change correlated negatively with the rate of increases in testosterone (r= -0.4, n=28, P < 0.05), LH (r= -0.5, n=26, P < 0.01) and inhibin B (r= -0.44, n=26, P < 0.05) levels.

CONCLUSIONS: In boys, peripheral MKRN3 levels decrease prior to clinical onset of puberty in an inverse association with circulating markers of HPG axis activity. Inhibition of estrogen biosynthesis has no effect on circulating MKRN3 levels.


Nothing to Disclose: TV, MH, KTV, LD, PM, TR

OR08-3 26365 3.0000 A Peripheral Makorin Ring-Finger Protein-3 (MKRN3) Levels in Boys Decline before the Clinical Onset of Puberty 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Pediatric Endocrinology Friday, April 1st 1:15:00 PM OR08 7755 11:45:00 AM Pediatric Endocrinology Oral

Christine M. Burt Solorzano*1, Eleanor G. Hutchens2, Su Hee Kim2, John C. Marshall2 and Christopher R. McCartney1
1University of Virginia, Charlottesville, VA, 2University of Virginia Health System, Charlottesville, VA


Pulsatile LH (GnRH) secretion is primarily nocturnal (sleep-related) in early female puberty; but daytime (while awake) LH pulse frequency gradually increases, and it exceeds nighttime frequency by late puberty. Mechanisms regulating these changes are unknown. In early pubertal girls, exogenous progesterone (P) can suppress daytime pulses, but it does not acutely suppress nighttime LH pulses. Since P suppresses daytime LH pulse frequency, changing daytime LH frequency across puberty may involve decreasing sensitivity of the daytime GnRH pulse generator to restraint by ambient P. As androgen excess impairs GnRH feedback inhibition to P, sensitivity to P may decrease with rising levels of testosterone (T). We aimed to assess further the hypothesis that relative increases in T across puberty are related to the increase in daytime LH pulses in girls.

We have studied 61 normal/overweight girls (Tanner I-V) via q 10 min sampling that included the 1900 to 2300 h time block—a period of wakefulness in all cases. LH pulse count for this 4 h time block was determined. Sex steroids were sampled q 60 min. Simple and partial Spearman rank correlations were used to assess relationships between LH pulse count while awake and sex steroid levels. Since both P and T may influence pulse frequency, and since both may change across puberty, we assessed the correlation between daytime LH pulse count and the molar ratio between P and free T (P-to-T molar ratio, with each hormone in pmol/L). Subjects were also analyzed in groups defined by daytime LH pulse counts: 0 (“none,” n=10), 1-2 (“some,” n=13), and ≥3 (“more,” n=38).

Among the sex steroids assessed, daytime LH pulse count correlated best with free T (R=0.68, p<0.0001), versus estradiol (R=0.40, p=0.001) and P (R=0.34, p=0.007). Daytime LH pulse count also exhibited a significant negative correlation with P-to-T molar ratio (R=-0.67, p<0.0001). When correcting for bone age (BA) or Tanner stage (TS), most correlations lost statistical significance—except for free T corrected for TS (R=0.40, p=0.002), P corrected for BA (R=-0.47, p=0.01), and P-to-T molar ratio corrected for either BA (R=-0.45, p=0.003) or TS (R=-0.49, p=0.01). Girls with no daytime LH pulses (“none”) tended to have lower free T levels (2.7 ± 0.5 pmol/L [mean ± SEM]) compared to “some” (19.5 ± 8.0) and “more” (22.8 ± 3.6) groups (p=0.057 and p<0.0001, respectively). Likewise, girls in the “none” group had higher P-to-T molar ratios (291 ± 49 pmol/L / pmol/L) compared to “some” (138 ± 24) and “more” (90 ± 12) groups (p=0.02 and p=0.003, respectively).

These results indicate that both increasing free T and decreasing P-to-T molar ratio correlate with daytime LH pulse counts, even when corrected for pubertal status. These data are consistent with the notion that rising T levels across puberty antagonize the ability of P to restrain daytime GnRH secretion, thus allowing a gradual increase in daytime LH frequency.


Nothing to Disclose: CMB, EGH, SHK, JCM, CRM

OR08-4 26893 4.0000 A Increasing Testosterone to Progesterone Ratio Is Associated with Evolution of Daytime GnRH Pulse Secretion during Puberty in Girls 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Pediatric Endocrinology Friday, April 1st 1:15:00 PM OR08 7755 11:45:00 AM Pediatric Endocrinology Oral

Janie Benoit*1, Kasiani Myers1, Stella M Davies1, Michael Grimley1, Sonata Jodele1, Pooja Khandelwal1, Javier El-Bietar1, Rebecca Marsh1, Adam Nelson1, Gregory Wallace1, Christopher Dandoy1, Pauline Daniels1, Abigail Pate1, Lesley Breech1, Holly Hoefgen1, Susan R Rose2 and Jonathan Christian Howell2
1Cincinnati Children's Hospital Medical Center, 2Cincinnati Children's Hospital Medical Center, Cincinnati, OH


Background:  Reduced intensity conditioning (RIC) preparative regimens are increasingly used for hematopoietic stem cell transplantation (HSCT) with the goal of maintaining therapeutic efficacy while limiting toxicity associated with more traditional, myeloablative regimens. Current understanding of late endocrine effects using RIC HSCT is extremely limited. While the risk of infertility after myeloablative HSCT is known to be high (>80%), there are currently no data regarding risk of infertility with RIC HSCT.

Objective:  Longitudinally evaluate gonadal function and fertility potential in a pediatric and young adult population after RIC HSCT.

Methods:  Children and young adults ≥ 1 year after a single RIC HSCT regimen were followed in our prospective cohort study to evaluate late effects of HSCT on the endocrine system and fertility potential. Subjects were evaluated for pubertal development and hormonal status, and semen analysis was obtained for specifically consented males.  All patients received RIC regimen prior to HSCT, with the majority receiving Fludarabine and Melphalan +/- Campath, and none had total body irradiation. Five of 14 patients (36%) had received prior chemotherapy for malignancy.  The median age at time of HSCT was 13.6 years (range 3.4-24.3). The average age at time of semen analysis following HSCT was 20.3 years (range 15.6-25.2), with a median time from HSCT of 5.6 years (range 1.9-10.0).

Results:  Preliminary results were obtained from 14 male subjects >11 years of age after RIC HSCT.  Of those, 11 (79%) were pubertal.  Of the 9 pubertal subjects with available laboratory testing, only one (11%) had abnormally elevated gonadotropins (LH and FSH).  Similarly, only one (11%) had an abnormally low testosterone level.  The remainder had normal testing for Tanner stage.  Interestingly, 4 of 9 (44%) had abnormally low inhibin B levels, suggestive of reduced fertility and Sertoli cell dysfunction.  Semen analysis was abnormal in all 8 subjects tested: azoospermia was diagnosed in 88% (7/8) and oligoteratospermia in 12% (1/8).  

Conclusion:  Our analysis suggests that, despite its potential benefits, RIC HSCT may be associated with a high risk of impaired spermatogenesis and infertility, similar to the risk associated with traditional, myeloablative HSCT. However, gonadotropins and testosterone appear to be normal in most young male subjects after RIC HSCT. Therefore, normal pubertal development does not ensure fertility potential following RIC HSCT.  Whereas our data suggest inhibin B levels can be useful for screening those at risk for infertility, semen analysis is a more reliable, definitive measure.  Risk of infertility should be included in counseling about RIC HSCT, and fertility preservation should be discussed and offered to these patients prior to HSCT if possible. Additional studies to confirm these data in a larger cohort are underway.


Nothing to Disclose: JB, KM, SMD, MG, SJ, PK, JE, RM, AN, GW, CD, PD, AP, LB, HH, SRR, JCH

OR08-5 26892 5.0000 A High Risk of Male Infertility after Hematopoietic Stem Cell Transplantation in Children and Young Adults Despite Reduced Intensity Conditioning 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Pediatric Endocrinology Friday, April 1st 1:15:00 PM OR08 7755 11:45:00 AM Pediatric Endocrinology Oral

Sjoerd D. Joustra*1, Charlotte A. Heinen2, Nadia Schoenmakers3, Marco Bonomi4, Bart EPB Ballieux1, Marc-Olivier Turgeon5, Daniel J. Bernard5, Eric Fliers2, A. S. Paul van Trotsenburg6, Monique Losekoot7, Luca Persani8, Jan Maarten Wit9, Nienke R. Biermasz9, Alberto M. Pereira9 and W. Oostdijk1
1Leiden University Medical Center, Netherlands, 2Academic Medical Center, University of Amsterdam, Amsterdam, Netherlands, 3University of Cambridge, 4Istituto Auxologico Italiano IRC, Trezzo Sull'Adda, Italy, 5McGill University, Montreal, QC, Canada, 6Academic Medical Center Amsterdam, Amsterdam, Netherlands, 7Laboratory for Diagnostic Genome Analysis, Leiden, Netherlands, 8Istituto Auxologico Italiano, milan, Italy, 9Leiden University Medical Center, Leiden, Netherlands


Context: Loss-of-function of the immunoglobulin superfamily member 1 (IGSF1) gene causes the X-linked IGSF1 deficiency syndrome, characterized by central hypothyroidism, delayed pubertal testosterone rise, adult macroorchidism, variable prolactin deficiency, and occasionally transient partial GH deficiency. Since our first reports, we discovered 20 new families with 18 new pathogenic IGSF1 mutations.

Objective: We aimed to share data on the largest cohort of patients with the IGSF1 deficiency syndrome to date and formulate recommendations for clinical management.

Methods: We collected clinical and biochemical characteristics of 69 male patients (35 children, 34 adults) and 56 female carriers of an IGSF1 mutation (3 children, 53 adults) from 30 unrelated families according to a standardized clinical protocol. At evaluation, 89% of boys were treated with levothyroxine, as were 44% of adult males, and 5% of females.

Results: Several additional symptoms were identified in male patients. Thyroid gland volume was small in 74%, birth weight was high in 25%, and head circumference was large in 20%. Late adrenarche was observed in patients with prolactin deficiency, and adult DHEA levels were decreased in 40%. In general, the timing of pubertal testicular growth was normal or even advanced, in contrast to a late rise in pubertal testosterone levels. Hypocortisolism was documented in 6 of 28 evaluated newborns, although cortisol levels were normal later on. Male patients' waist circumference was increased in 60%, but blood lipids were normal. Female carriers showed low FT4 and low-normal FT4 in 18% and 60%, respectively, delayed age at menarche in 31%, mild prolactin deficiency in 22%, increased waist circumference in 57%, and a negative correlation between FTconcentrations and metabolic parameters. Based on the large number of patients identified in only three years, IGSF1 deficiency represents the most common genetic cause of relatively isolated central hypothyroidism. 

Conclusion: IGSF1 deficiency syndrome represents the most common genetic cause of central hypothyroidism and is associated with multiple other characteristics. The results allow the formulation of recommendations for mutational analysis, endocrine work-up, and long-term care.


Nothing to Disclose: SDJ, CAH, NS, MB, BEB, MOT, DJB, EF, ASPVT, ML, LP, JMW, NRB, AMP, WO

OR08-6 24338 6.0000 A The IGSF1 Deficiency Syndrome: Lessons from an Extensive Case Series 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Pediatric Endocrinology Friday, April 1st 1:15:00 PM OR08 7755 11:45:00 AM Pediatric Endocrinology Oral

Lucia Speroni*1, Maria Voutilainen2, Marja L Mikkola2, Skylar A Klager1, Cheryl M Schaeberle1, Carlos Sonnenschein1 and Ana M Soto1
1Tufts University, Boston, MA, 2Institute of Biotechnology, University of Helsinki, Helsinki, Finland


Fetal exposure to Bisphenol-A (BPA) causes alterations in mammary gland development increasing the risk of breast cancer later in adulthood (1). At embryonic day (E) 18, the mammary stroma shows increased adipocyte differentiation, altered organization of collagen fibers, decreased deposition of tenascin-C, while the epithelial tree is enlarged and the formation of ductal lumen is delayed (2). Although estrogen receptors are present in the stroma of the fetal mammary gland, it is yet unknown whether these effects are directly mediated by BPA and/or whether BPA is acting indirectly via the hypothalamic-pituitary-ovarian axis.

To address the question of whether BPA acts directly, we utilized an ex vivo culture method of the fetal mammary gland (3). In this method, the direct action of estrogen and estrogen-mimics can be tested during E14 to 19, a critical window of exposure. This method allows for the direct observation of development as it occurs.

Mammary buds of CD1 mice were dissected at E14 and cultured for 5 days. The explants were exposed to BPA or 17β-estradiol (E2). Morphometric analysis was performed on the explant whole-mounts and markers of epithelial and mesenchymal development were detected by immunofluorescence.

We show that BPA exerts a direct effect on the fetal mammary gland. Exposure of the explants to 10-9M BPA significantly increased ductal growth (89711 ± 6836 µm2; p=0.028; n=20) while 10 -6M BPA decreased it (33518 ± 3761 µm2; p=0.006; n=9) compared to controls (67615 ± 8806 µm2; n=15). Epithelial growth was significantly diminished in mammary buds exposed to 10-9 (48337 ± 5200 µm2; p=0.005; n=16), 10-11 (49441 ± 4604 µm2; p=0.000; n=39) and 10-13M (54414 ± 6404 µm2; p=0.031; n=15) E2 compared to controls (76802 ± 6305 µm2; n=43); lower doses had no effect.

Our findings show that fetal mammary gland development is altered by direct action of estrogenic compounds. BPA shows a non-monotonic dose response curve whereby low dose increases ductal development and high doses inhibit it. Moreover, increased ductal development was observed at doses comparable to those producing similar effects in vivo (2). In contrast, estradiol resulted in a monotonic inhibition of ductal growth. Finally, in addition to its usefulness to the understanding of the hormonal regulation of mammary gland development, this ex vivo culture system could serve as a bioassay to test the numerous new chemicals that are synthesized each year and released into the environment without proper assessment of hormonal action on critical targets like the mammary gland.


Nothing to Disclose: LS, MV, MLM, SAK, CMS, CS, AMS

OR04-1 26102 1.0000 A An Ex-Vivo Organ Culture Shows That BPA Directly Affects the Developing Mammary Gland 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Endocrine Disruptors Friday, April 1st 1:15:00 PM OR04 7761 11:45:00 AM Endocrine Disrupting Chemicals Oral

Ana Cheong*1, Xiang Zhang1, Yuk-Yin Cheung1, Jing Chen1, Mario Medvedovic1, Gail S. Prins2 and Shuk-Mei Ho1
1University of Cincinnati College of Medicine, Cincinnati, OH, 2University of Illinois-Chicago, Chicago, IL


Exposure to environmental xenoestrogens such as estradiol benzoate (EB) and bisphenol A (BPA) is associated with increased prostate cancer (PCa) risk. In humans, higher estradiol or urinary BPA levels are detected in PCa patients, but  the mechanistic link to PCa is unclear. We previously reported that exposing rat neonates to 2,500 µg/kg BW EB or 10 µg/kg BW BPA on postnatal day (PND) 1, 3, and 5 persistently upregulated phosphodiesterase type 4 variant 4 in PND10, 90, and 200 prostates through hypomethylation, which was associated with increased susceptibility to estradiol (E2)-driven carcinogenesis with aging. This initial data was proof of principle for epigenetic reprogramming as an underpinning for increased PCa risk.   In the present study, we sought to identify multiple epigenetically reprogrammed genes in the PND90 dorsal prostate as a function of neonatal exposures that prime the gland for E2-driven carcinogenesis initiated at day 90.  We performed methylated CpG island recovery assay-assisted methylation promoter array profiling and identified 111 EB-associated and 86-BPA associated genes. These genes were commonly associated with “cancer”, “cell-to-cell signaling and interaction, cell-mediated immune response, cellular growth and proliferation” and “nucleic acid metabolism, small molecule biochemistry, molecular transport”. To understand the underlying EB/BPA mechanisms, we used a 750 bp sliding window approach with stringent criteria and selected the nine most differentially methylated genes for studying the correlation of their promoter methylation status and gene expression in  dorsal prostates using bisulfite sequencing and qPCR analyses. We found seven out of nine genes had an inverse correlation between promoter methylation pattern and gene expression in the PND90 prostate. To determine if the differential methylation and gene expression patterns persist in PND200 when carcinogenesis was apparent, the top four genes studied in PND90 tissues were analyzed in PND200 dorsal prostates. Although some methylation patterns were shifted in PND200 and upon T+E2 treatment from PND90-200, gene expression was persistent. The fact that these differentially methylated promoters were associated with higher PIN incidence in our rodent model and with poor survival of PCa patients revealed that the epigenetically reprogramming effects of early-life EB or BPA exposure is genome-wide and is associated with higher PCa risk.


Nothing to Disclose: AC, XZ, YYC, JC, MM, GSP, SMH

OR04-2 26118 2.0000 A Epigenetic Reprogramming of Rat Prostate By Neonatal Exposure to Estradiol Benzoate and Bisphenol-a 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Endocrine Disruptors Friday, April 1st 1:15:00 PM OR04 7761 11:45:00 AM Endocrine Disrupting Chemicals Oral

Sheryl E Arambula*1, Scott M Belcher2, Stephen D Turner3, Keith L Gonzales4 and Heather B Patisaul1
1North Carolina State University, Raleigh, NC, 2University of Cincinnati, Cincinnati, OH, 3University of Virginia School of Medicine, Charlottesville, VA, 4Stirplate.io, Stirplate Inc., New York, NY


Bisphenol A (BPA) is an endocrine disrupting, high volume production chemical found in a wide variety of products including plastics, epoxy resins and thermal paper receipts. Human exposure is nearly ubiquitous, but higher in children than adults. Concern has been raised that exposure, even at doses below the current reference dose of 50 µg/kg bw/day can disrupt the developing brain. Studies by us and others have shown that developmental BPA exposure can alter the sex-specific expression of estrogen responsive genes in the neonatal rodent brain, including estrogen receptors, but a clear dose response curve remains uncertain and the full range of transcriptional effects remains poorly characterized, particularly in extra-hypothalamic regions.  To address this, the present studies were conducted as part of the CLARITY-BPA (Consortium Linking Academic and Regulatory Insights on BPA Toxicity) program and tested the hypothesis that prenatal BPA exposure induces transcriptional changes in the neonatal rat hypothalamus (critical for reproductive and affective behaviors) and hippocampus (critical for spatial navigation and memory). NCTR Sprague-Dawley dams were orally gavaged from gestational day 6 until parturition to BPA (2.5, 25, 250, 2500, or 25000 µg/kg bw/day), ethinyl estradiol as a reference estrogen (0.05 or 0.5 µg/kg bw/day), or vehicle. The brains of postnatal day 1 pups (n = 10 per sex per group) were flash frozen and the regions of interest dissected via micropunch. Total RNA was isolated from a subgroup (n = 4 per sex from vehicle, BPA 2.5 and BPA 2500 groups) and assessed via RNAseq. No effect of BPA was detected on the hippocampal transcriptome. In the hypothalamus, 34 genes were identified as putatively impacted in the male BPA 2500 group. Subsequent qRT-PCR on a larger subset (n = at least 5 per sex per group from all groups) revealed significant, sex-specific BPA-related effects on the hypothalamic expression of estrogen receptor α (ESR1), estrogen receptor β (ESR2), oxytocin (OXT), and GABA vesicular transporter (SLC32A1) at doses as low as 2.5 µg/kg bw/day, and hippocampal expression of OXT and ESR2. These data provide supporting evidence for the hypothesis that BPA exposure has the potential to alter the sex-specific transcriptome of the developing brain, even at doses below the current reference dose.


Nothing to Disclose: SEA, SMB, SDT, KLG, HBP

OR04-3 25513 3.0000 A Prenatal Exposure to Bisphenol a Alters Hypothalamic and Hippocampal Gene Expression in Neonatal Rats of Both Sexes 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Endocrine Disruptors Friday, April 1st 1:15:00 PM OR04 7761 11:45:00 AM Endocrine Disrupting Chemicals Oral

Shannon D. Whirledge*1, Matthew Quinn2 and John A Cidlowski3
1NIEHS, Durham, NC, 2NIEHS, RTP, NC, 3NIEHS/NIH, Research Triangle Park, NC


Developmental exposure to environmental estrogens results in many long-term consequences to the female reproductive tract. In rodent models, neonatal exposure to the phytoestrogen genistein results in infertility due to the inability of the female reproductive tract to support pre-implantation embryo development and implantation. The uterus is permanently altered by early exposure to genistein, including significant changes in the expression of immune response genes important in early pregnancy. Glucocorticoids are primary stress hormones with potent immunomodulatory actions. We have recently discovered that GR signaling within the uterus is critical for pregnancy. Uterine GR KO mice display a profound sub-fertile phenotype, which includes dysregulation of immune response genes and altered immune cell recruitment during early pregnancy. Disruption of GR signaling in the uterus by neonatal genistein exposure may underlie the infertility phenotype. To determine if the phytoestrogen genistein alters GR signaling in the mouse uterus, C57Bl/6 pups were injected subcutaneously on postnatal days 1-5 with 50 mg/kg genistein or vehicle (corn oil). Mice were then treated on Day 5 or as adults with the synthetic glucocorticoid dexamethasone, and gene expression was analyzed. The response to dexamethasone was equivicable in the uterus of genistein and vehicle treated mice at day 5. In contrast, neonatal genistein exposure resulted in blunted expression of Gilz and Fkbp5, two classic glucocorticoid-responsive genes, in the adult uterus. To determine if this difference in hormone responsiveness was reflected at the whole genome level, microarray analysis was performed on adult mice that were exposed to neonatal genistein or vehicle. The robust transcriptional response to glucocortcoids is significantly impaired by neonatal genistein exposure. In the uterus of adult control mice, 3382 genes are significantly regulated by dexamethasone treatment, but only 2353 genes are significantly regulated in the uterus of mice exposed to genistein during the neonatal period. There are also significant changes in the basal expression of many glucocorticoid target genes. These studies suggest that developmental exposure to environmental estrogens can lead to persistent alterations in glucocorticoid signaling in the adult uterus. These changes may be mediated by modifications to the chromatin architecture, since we discovered that neonatal exposure to genistein results in aberrant expression of DNA methyltransferases and histone deacetylases. Impairment to glucocorticoid signaling by early exposure to genistein results in defects in ability of dexamethasone to regulate the biological functions of the uterus. Thus disruption of glucocorticoid signaling in the uterus via environmental estrogen exposure may be directly responsible for the inability of the uterus to support implantation.


Nothing to Disclose: SDW, MQ, JAC

OR04-4 27109 4.0000 A Neonatal Genistein Exposure Permanently Alters Glucocorticoid Signaling in the Mouse Uterus 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Endocrine Disruptors Friday, April 1st 1:15:00 PM OR04 7761 11:45:00 AM Endocrine Disrupting Chemicals Oral

Lindsey S. Treviño*1, Charles E Foulds2, Philip F Lavere2, Jun Qin2, Bert W O'Malley2 and Cheryl Lyn Walker1
1Texas A&M Health Science Center, Houston, TX, 2Baylor College of Medicine, Houston, TX


Obesity has become a major health issue worldwide, and both human and experimental animal data point to a role for early-life exposures to endocrine-disrupting chemicals (EDCs) in promoting obesity later in adulthood.  We have utilized a model obesogen, tributyltin (TBT) to explore how engagement of nuclear receptors (NRs), primarily PPARγ/RXRα, by TBT can induce developmental reprogramming in the liver.  Our hypothesis is that select EDCs may alter adult liver function upon exposures during critical developmental windows by altering NR-coregulator (CoR) complex formation to perturb “classical” NR programming of the epigenome.

 We find that exposure of the developing liver to TBT, or treatment of liver cells in vitro, alters the expression of key genes involved in energy homeostasis, including Shp and Acox.  Altered expression of these genes correlates with increased H3K4me3, indicating that TBT-induced changes in transcription are associated with altered histone methyltransferase (HMT) activity.  To test the hypothesis that TBT engagement of PPARγ/RXRα is directly responsible for altered HMT recruitment/activity, we have developed a DNA pulldown assay employing recombinant PPARγ and RXRα proteins incubated with rat liver FAO cell nuclear extract (NE), DMSO vehicle or TBT, and a biotinylated Shp gene promoter containing an evolutionarily conserved PPAR/RXR binding site. After pulldown with streptavidin beads, bound CoR proteins were detected by mass spectrometry (MS). From the MS data, we identified proteins involved in chromatin remodeling, tethering/looping, and histone modifications (histone acetyltransferases (HATs) and HMTs) that were enriched at least 2-fold with TBT. Similar CoR recruitments were seen using a biotinylated 3xPPRE template derived from the rat Acox promoter.

Importantly, TBT recruited p300, DPY30, a core component of SET/MLL HMT complexes, and SRC-2 to PPREs in an RXR-dependent manner. Consistent with this finding, the RXR inhibitor HX531 reduced TBT-induced recruitment of these three CoRs from HeLa cell NE to 3xPPREs. Combined these data suggest that specific CoRs are recruited to target genes by TBT binding to RXR, and point to a model where TBT binding to the PPAR/RXR heterodimer recruits distinct histone “modifying” CoRs to “write” histone marks that will persist to reprogram TBT-target genes in the adult liver to a metabolic setpoint favoring obesity.

* LST and CEF contributed equally to this work.


Nothing to Disclose: LST, CEF, PFL, JQ, BWO, CLW

OR04-5 27151 5.0000 A Mechanistic Insights into How a Model Obesogen, Tributyltin (TBT), Affects Peroxisome Proliferator Activator Gamma (PPARg) and Retinoid X Receptor Alpha (RXRa) Coregulator Complexes on Target Genes 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Endocrine Disruptors Friday, April 1st 1:15:00 PM OR04 7761 11:45:00 AM Endocrine Disrupting Chemicals Oral

Sören Verstraete*1, Ilse Vanhorebeek1, Adrian Covaci2, Fabian Güiza3, Govindan Malarvannan2, Philippe G Jorens4 and Greet Van den Berghe3
1KU Leuven, Leuven, Belgium, 2University of Antwerp, Antwerp, Belgium, 3Katholieke Universiteit Leuven, Leuven, Belgium, 4Antwerp University Hospital, Edegem, Belgium



Background: The long-term legacy of pediatric critical illness comprises a severe and unexplained attention deficit.1 Phthalates, which may be neurotoxic, are used to soften plastic indwelling medical devices and can leach into the circulation.2, 3

Hypothesis: We hypothesized that in children treated in the pediatric intensive care unit (PICU), circulating phthalates leaching from indwelling medical devices contribute to their long-term attention deficit.

Methods: Circulating plasma concentrations of di(2-ethylhexyl)phthalate (DEHP) metabolites were quantified in 100 healthy children and 449 children who had been treated in PICU and were neurocognitively tested 4 years later. In a development patient cohort (N=228), a multivariable bootstrap study identified stable thresholds of exposure to circulating DEHP metabolites above which there was an independent association with worse neurocognitive outcome. Subsequently, in a second patient cohort (N=221), the observed independent associations were validated.

Results: Plasma concentrations of DEHP metabolites, that were virtually undetectable [0.029(0.027-0.031) µmol/l] in healthy children, were 4.41(3.76-5.06) µmol/l in critically ill children upon PICU admission (P<0.001). Plasma DEHP metabolite concentrations decreased rapidly but remained 18-times elevated until PICU discharge (P<0.001). After adjusting for baseline risk factors and duration of PICU stay, and further for PICU complications and treatments, exceeding the potentially harmful threshold for exposure to circulating DEHP metabolites was independently associated with the attention deficit (all P≤0.008) and impaired motor coordination (all P≤0.02). The association with the attention deficit was confirmed in the validation cohort (all P≤0.01). This phthalate exposure effect explained half of the attention deficit in post-PICU patients.

Conclusions and Relevance: Exposure to phthalates leaching from indwelling medical devices used for intensive medical care in children was independently and robustly associated with their long-term attention deficit. Development of alternative plasticizers for this application may be indicated.


Nothing to Disclose: SV, IV, AC, FG, GM, PGJ, GV

OR04-6 23885 6.0000 A Circulating Phthalates during Critical Illness in Children and Their Long-Term Attention Deficit Legacy: An Association Study of a Development and a Validation Cohort 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Endocrine Disruptors Friday, April 1st 1:15:00 PM OR04 7761 11:45:00 AM Endocrine Disrupting Chemicals Oral

Nozomi Takahashi*1, Miyuki Harada1, Yasushi Hirota1, Osamu Yoshino2, Osamu Hiraike1, Tomoyuki Fujii1 and Yutaka Osuga1
1The University of Tokyo, Tokyo, Japan, 2The University of Toyama, Toyama, Japan


Ovarian hyperstimulation syndrome (OHSS) is characterized by increased vascular permeability (VP), which is mediated by vascular endothelial growth factor (VEGF) overexpressed in granulosa-lutein cells (GLCs) (1). Recent studies reveal that the spliced form of X-box-binding protein 1 [XBP1(S)], an unfolded protein response (UPR) transcription factor activated by endoplasmic reticulum (ER) stress, regulates VEGF expression (2). In a previous study, we demonstrated that the granulosa cells of growing follicles express XBP1(S) mRNA, which is dependent on follicular stage: specifically, XBP1(S) mRNA is expressed in granulosa cells of the follicles in later stages (large secondary, antral, and pre-ovulatory) (3). We hypothesized that XBP1(S) induced in granulosa cells of follicles in later stages modulates LH/hCG-induced VEGF expression and plays a role in the pathophysiology of OHSS. To test this hypothesis, we first examined in vitro effect of ER stress on VEGF mRNA expression and protein secretion in cultured human GLCs by real-time quantitative PCR (qPCR) and ELISA, respectively. Treatment of human GLCs with tunicamycin (Tm), an ER stress inducer, increased VEGF and XBP1(S) mRNA expression. In GLCs treated with Tm + hCG, the VEGF mRNA level and protein secretion was higher, with a concomitant induction of XBP1(S) mRNA, than that in GLCs treated with hCG alone. To examine the intermediary role of XBP1(S) in the upregulation by Tm of hCG-induced VEGF production, we knocked down XBP1(S) mRNA by RNA interference. XBP1(S) expression was knocked down by 60%, and VEGF mRNA expression was reduced 20%. Treatment with tauroursodeoxycholic acid (TUDCA), an ER stress inhibitor already in clinical use, significantly reduced hCG-induced VEGF mRNA expression and protein secretion in Tm-pretreated GLCs to almost the same level as in GLCs without Tm treatment. Finally, we examined the in vivo effect of TUDCA on OHSS pathology in well-established rat model. OHSS was induced in three-week-old female wistar rats by injections of pregnant mare serum gonadotropin (PMSG) and hCG, and TUDCA was injected intraperitoneally during PMSG-hCG treatment. Forty-eight hours after hCG injection, VP was measured and ovaries were collected. Ovarian weight, the number of corpora lutea (CL), and VP were significantly higher in the OHSS group than in the control group. TUDCA treatment suppressed the increase in VP in the OHSS group, without affecting ovarian weight or the number of CL. The increase in expression of VEGF mRNA in the ovary observed in the OHSS group was downregulated by TUDCA treatment. Our results suggested that ER stress upregulates the hCG-induced VEGF production in human GLCs. TUDCA prevents development of OHSS by reducing VEGF production in granulosa cells. Thus, given its stimulatory effect on VEGF production in granulosa cells, ER stress represents a novel therapeutic target for preventing OHSS.


Nothing to Disclose: NT, MH, YH, OY, OH, TF, YO

OR09-1 24018 1.0000 A Endoplasmic Reticulum Stress Induces Vascular Endothelial Growth Factor Production in Granulosa Cells: Implications for a Novel Therapeutic Approach for Ovarian Hyperstimulation Syndrome 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Reproductive Endocrinology Friday, April 1st 1:15:00 PM OR09 7767 11:45:00 AM Role of Growth Factor Signaling in Fertility and Pregnancy Oral

Sha Li* and Mark Stephen Roberson
Cornell College of Veterinary Medicine, Ithaca, NY


The angiogenic factor human placental growth factor (hPGF) is abundantly expressed in placental trophoblasts. Low serum levels of hPGF have been associated with preeclampsia (PE), which is the leading cause for maternal and fetal morbidity and mortality. Further, hPGF has been proposed as a biomarker for PE. Despite the recognition of its pathophysiological importance during pregnancy, mechanisms of hPGF transcriptional regulation remain largely unknown.  Limited published data suggests that the hPGF promoter is regulated by Glial cell missing 1 (GCM1) in human choriocarcinoma cell lines. Further, the transcription factor Distal-less 3 (Dlx3) is a critical regulator of PGF gene expression in mouse trophoblasts based upon microarray analysis of the Dlx3 null placenta. Here we propose that DLX3 regulates hPGF expression in conjunction with GCM1 in human placental trophoblasts, based on the following experimental observations. Chromatin immunoprecipitation (ChIP) studies in choriocarcinoma cells (JEG3) revealed the binding of DLX3 at the promoter region of hPGF gene (p<0.05). Secondly, overexpression of DLX3 transactivated hPGF promoter in a luciferase reporter system (p<0.05), whereas GCM1 overexpression was observed to be a much stronger transactivator (p<0.05). Interestingly, co-expression of DLX3 and GCM1 lead to an antagonist effect on hPGF (p<0.05), suggesting a modulatory role of DLX3 together with GCM1 in regulation of hPGF. RT-qPCR and western blot studies in JEG3 cells showed a consistent patterns of changes in endogenous hPGF mRNA and protein induced by DLX3 and GCM1, respectively and combinatorially (p<0.05). Knock down of either DLX3 (p<0.05) or GCM1 (p<0.05) alone significantly reduced endogenous hPGF expression, while a combinational knockdown of the two regulators had an additive effect on hPGF downregulation (p<0.05). Based upon deletion mutagenesis studies, we were able to identify a ~50 nucleotide sequence upstream the transcription start site of the hPGF gene that is critical for basal expression and the regulatory responses to DLX3 and GCM1.  In addition, ChIP studies detected the enrichment of both GCM1 and DLX3 on this cis element (p<0.05), suggesting an interaction of DLX3 and GCM1 in this loci. To better understand this mechanism, GAL4/UAS one-hybrid studies were carried out using Gal4-GCM1 as bait.  These studies supported the conclusion that these two transcription factors functionally interact to mediate the antagonist effect on hPGF expression. Our study is the first to describe regulatory cross-talk between DLX3 and GCM1, and their cooperative function on hPGF gene regulation. These studies potentially provide fundamental insight into molecular events that occurs in human trophoblasts during pregnancy, and help to understand mis-regulation of transcriptional networks in the placenta associated disease conditions including PE.


Nothing to Disclose: SL, MSR

OR09-2 26954 2.0000 A Human Placental Growth Factor Is Regulated By a Cooperation of DLX3 and GCM1 in Human Trophoblast Cells 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Reproductive Endocrinology Friday, April 1st 1:15:00 PM OR09 7767 11:45:00 AM Role of Growth Factor Signaling in Fertility and Pregnancy Oral

Jessica L Brown*1, Jennifer L Sones1, Ulrich Boehm2 and Mark Stephen Roberson1
1Cornell College of Veterinary Medicine, Ithaca, NY, 2University of Saarland School of Medicine, Homburg, Germany


Extracellular-signal-regulated kinase (ERK) 1 and 2 signaling is an important regulatory pathway within the hypothalamic-pituitary-gonadal axis. To understand the specific role of ERK1/2 signaling in gonadotrope cells in the anterior pituitary, we created conditional knockout (ko) mice lacking ERK1 (null allele) and ERK2 (floxed allele) in cells expressing the GnRH receptor (GnRHR). As expected, conditional inactivation of ERK1/2 signaling in the double KO (ERKdko) animals resulted in loss of luteinizing and follicle-stimulating hormones (LH/FSH) and subsequent loss of ovulation.

Exogenous administration of FSH and LH rescued anovulation and lead to subsequent pregnancy in approximately one-third of the ERKdko animals. However, litter sizes from ERKdko mice were significantly smaller than controls (2.4±0.29 vs. 9±1 pups, n=7, n=3, respectively, p<0.01). Furthermore, 15/17 pups from ERKdko mothers recovered were stillborn, and the two live pups died within hours of birth. No ERKdko pups were alive by postnatal day 3 (0% vs. 85.7± 11.7% survival in control litters, n=7, n=3, respectively, p<0.01). Gestation length was significantly longer in ERKdko animals (23.9±0.55 days vs. 20.25±0.25 days, n=5 and n=4 respectively, p<0.01), and parturition lasted significantly longer (2.25±0.42 days vs. 1 day in control animals, n=4 each, p<0.05).

To dissect the mechanisms underlying this profound reproductive phenotype, we started to examine the placentas for potential GnRHR expression during gestation. We performed qRT-PCR on C57/Blk6 mouse implantation sites/placentas from embryonic day (e)5.5 (n=4), e10.5 (n=4), e12.5 (n=3), and e18.5 (n=1) and compared them to pituitary GnRHR mRNA expression levels. Strikingly, we found that placental GnRHR mRNA levels were comparable to those in the pituitary at e5.5 and e10.5, and 9-fold greater in the placenta at e18.5. These studies suggest that GnRHR-mediated Cre expression in the placenta may effectively inactivate the floxed ERK1/2 target genes in this tissue. We speculate that loss of ERK signaling in the placenta of ERKdko animals causes alterations of the maternal/fetal interface leading to disruption of initiation and/or progression of parturition, resulting in fetal mortality. Further, these studies reveal a potentially important model of placental-specific Cre recombinase.


Nothing to Disclose: JLB, JLS, UB, MSR

OR09-3 26531 3.0000 A Loss of ERK1/2 Signaling in Cells Expressing the GnRH Receptor Causes Prolonged Gestation, Dystocia, and Fetal Death 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Reproductive Endocrinology Friday, April 1st 1:15:00 PM OR09 7767 11:45:00 AM Role of Growth Factor Signaling in Fertility and Pregnancy Oral

Oran Yakubovsky*1, Irina Gurt2, Einav Cohen-Kfir1, Hanna Artsi2, Gera Verbun2 and Rivka Dresner-Pollak2
1Hebrew University - Hadassah Medical School, Jerusalem, Israel, 2Hadassah-Hebrew University Medical Center, Jerusalem, Israel


Menopause is a period of accelerated aging in women. It is associated with weight gain, blood pressure elevation and an increased risk of developing cardiovascular disease and dementia. Sirtuin1 (Sirt1), a NAD+-dependent deacetylase, is a cellular energy sensor and a major player in metabolism and aging. Sirt1 over-expression confers protection against diabetes, obesity and dementia in mice. We asked if Sirt1 plays a role in the metabolic changes that occur with the menopausal transition. We have previously reported that Sirt1 is decreased in bone in ovariectomized (OVX) mice (1).

To test the hypothesis that Sirt1 level or activity is altered with OVX, 9-week-old C57BL/6 female mice (n=10/group) were subjected to OVX or SHAM operation, and were left untreated for 6 weeks. OVX and SHAM mice were sacrificed 1 and 6 weeks post operation. The remaining OVX mice were treated for 6 weeks with either sc daily 10mg/kg 17-β estradiol (E2), SRT3025, a Sirt1 activator, administered by gavage kindly provided by Sirtris/GSK, at 100 mg/kg/day once daily or a vehicle. SHAM mice were left untreated. Mice were kept on standard chow diet, weighed weekly, and serum was collected 1,6,12 weeks post operation. Upon sacrifice uterine weight was determined to ensure a successful OVX, livers and brains were collected. In vitro experiments were conducted in a model cell line.

OVX induced a significant weight gain of 27% and a 61% decrease in uterine weight over the 12 week period. Treatment with SRT3025 blunted OVX-induced weight gain (1.9% vs 4.6% in SRT3025-treated and untreated OVX mice, respectively), whereas E2-treated mice had a weight gain of 3.9%. Uterine weight was similar in OVX untreated and OVX SRT3025-treated mice (36.3±21.04 mg and 45.7±22.0 mg, respectively vs. 97.6±19.5 mg in E2-treated mice), suggesting no uterine estrogen-like effect. Strikingly, a dramatic decrease of 40% in Sirt1 protein level was observed 6 weeks post operation in liver and brain obtained from OVX compared to SHAM mice. Liver Sirt1 mRNA expression was significantly elevated by 1.8 fold in OVX compared to SHAM mice. To elucidate underlying mechanisms, C3H10T1/2 cells were exposed to 10-100nM E2, 1-10uM fulvestrant or serum derived from OVX and SHAM mice 1 week post operation. While no effect in Sirt1 protein level was detected in E2- and fulvestrant-treated cells, serum derived from OVX mice induced a marked decrease in Sirt1 expression, suggesting that OVX-related humoral factors influence Sirt1 expression.

In conclusion, reduced Sirt1 is a possible contributor to OVX-induced weight gain. Reduced Sirt1 appears to result from indirect effects of estrogen withdrawal. Pharmacologic activation of Sirt1 blunted OVX-associated weight gain without inducing an undesired effect of increased uterine weight. Future studies are needed to evaluate the effects of Sirt1 activation on menopause-related metabolic derangements and co-morbidities.


Nothing to Disclose: OY, IG, EC, HA, GV, RD

OR09-4 24709 4.0000 A Menopause Is a Low Sirtuin1 State: The Ovariectomized Mouse Model 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Reproductive Endocrinology Friday, April 1st 1:15:00 PM OR09 7767 11:45:00 AM Role of Growth Factor Signaling in Fertility and Pregnancy Oral

Rachel A Ross*1, Caroline A. Maguire2, Anne M. J. Verstegen3, Ursula B. Kaiser4, Bradford Barr Lowell5 and Victor M. Navarro6
1Beth Israel Deaconess Medical Center, Massachusetts General Hospital and Harvard Medical School, Boston, MA, 2Brigham and Women's Hospital/Harvard Med School, Boston, MA, 3Beth Israel Deaconess Medical Center and Harvard Medical School, 4Brigham and Women's Hospital/Harvard Medical School, Boston, MA, 5Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA, 6Harvard Medical School and Brigham and Women's Hospital, Boston, MA


Leptin is known to play a critical role in control of metabolism and reproduction, but the mechanisms by which this occurs are not yet understood. Pituitary adenylate cyclase activating polypeptide (PACAP) is a neuromodulator implicated in human anxiety, feeding and reproductive behavior that colocalizes with glutamate in the brain. PACAP whole body knockout mice display decreased fertility. PACAP stimulates LH release directly at the level of the gonadotrophs in the pituitary and increases sensitivity to GnRH, but the origin of the neuropeptide, or its role at the hypothalamic level, remains unknown. Centrally, high expression of PACAP (Adcyap1) is found in the ventral premamillary nucleus of the hypothalamus (PMV) and the ventromedial hypothalamus (VMH), both regions known to be involved in leptin-related control of puberty and fertility, though the role of PACAP has not been investigated there. To investigate the role that central PACAP plays in leptin-driven metabolism and reproduction, we created lox-PACAP mice that possess loxP sites flanking the 2nd exon of the PACAP gene, allowing for deletion of functional PACAP in the presence of cre-recombinase. The PACAP-lox mice were bred with LepRb-cre mice, which express cre-recombinase under control of the promoter for the long form of the leptin receptor (LepRb), which is restricted to the brain. Mice were subjected to assays for energy homeostasis and fertility, and compared to littermate controls (PACAPfl/fl). Conditional knock out females lacking PACAP in LepRb-expressing neurons show slight protection against body weight gain seen in control animals when subjected to high fat diet. Interestingly, they have significantly delayed puberty onset, as determined by delayed vaginal opening and first estrus. Females show irregular (prolonged) estrous cycles and decreased litter size. There is no change in male body weight, onset of puberty, or fertility. Dual-fluorescent immunohistochemistry and in situ hybridization revealed co-localization of LepR activity with PACAP expression in the ventral premammillary nucleus and the central part of the ventromedial nucleus of the hypothalamus. Therefore, in females, deletion of PACAP from leptin receptor expressing neurons in these two discrete regions of the hypothalamus involved in the regulation of the gonadotropic axis leads to fertility impairments similar to those seen in whole body PACAP knockouts. Based on these findings, we propose a new, sex specific role for the PACAP-containing leptin-responsive neurons of the hypothalamus in signaling nutritional state to regulate GnRH release, indicating that a subset of glutamatergic neurons may be involved in fine-tuning leptin’s action on reproductive function.


Nothing to Disclose: RAR, CAM, AMJV, UBK, BBL, VMN

OR09-5 25379 5.0000 A The Effect of PACAP on Fertility Is Relayed through a Subset of Hypothalamic Leptin Receptor Expressing Neurons in the Female Mouse 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Reproductive Endocrinology Friday, April 1st 1:15:00 PM OR09 7767 11:45:00 AM Role of Growth Factor Signaling in Fertility and Pregnancy Oral

Samantha Weaver*, Laura L Hernandez and Chad Vezina
University of Wisconsin-Madison, Madison, WI


Selective serotonin reuptake inhibitors (SSRI) are the class of antidepressants most commonly prescribed during pregnancy and lactation. Chronic use of SSRI has been demonstrated to result in decreased bone mineral density and increased fracture risk across all ages and sexes. Lactation is also characterized by increased bone resorption to free calcium stores available to move into milk. The action of serotonin on the mammary gland plays a prominent role in initiating hormonal cascades responsible for bone demineralization during lactation. We hypothesize that the use of SSRI during lactation causes excessive bone resorption from which women may be unable to recover. High doses of folic acid may work to reverse bone loss by mediating serotonin’s action on the mammary gland. To examine the effects of SSRI and folic acid administration on bone loss during lactation, we treated C57B6/J dams with the SSRI fluoxetine hydrochloride (20 mg/kg) or saline daily beginning on d 13 of pregnancy through d 10 of lactation. Beginning two weeks before breeding, mice were also exposed to either a breeder diet, or a diet supplemented with 20 mg/kg of folic acid, creating the following groups: breeder diet/fluoxetine (BF), breeder diet/saline (BS), folic acid/fluoxetine (FF), and folic acid/saline (FS). Milk yield was recorded daily using the weigh-suckle-weigh method. Blood samples were taken before beginning injections, and on d 1 and d 10 of lactation. On d 10 of lactation, mice were sacrificed and mammary glands and femurs were collected. There was an overall treatment effect on circulating serotonin levels (P<0.05). On d 1 of lactation, BS mice had higher serum serotonin levels than BF mice (7,795 ± 1,530 ng/mL vs. 743 ± 80) and FS mice had elevated serotonin compared to FF mice (13,835 ± 4,135 ng/mL vs. 1,378 ± 870). A similar pattern was observed on d 10 of lactation. BF mice produced significantly more milk than all other groups throughout the entire lactation (P<0.0001; 0.16 ± 0.09 g/pup vs. 0.09 ± 0.05). BF mice had higher levels of circulating calcium than all other groups on d 1 of lactation (P<0.05), but not on d 10. Finally, mRNA expression of serotonin transporter SERT in mammary gland and bone-building osteocalcin in femur was higher in BS and FS groups compared to BF and FF groups, respectively (P<0.05), and expression of bone breakdown M-CSF in femur was increased in BF and FF groups compared to BS and FS mice (P<0.05). In femur, folic acid tended to reverse the effects of SSRI on osteocalcin and M-CSF expression. Taken together, these data suggest that SSRI use during pregnancy and lactation induces changes in the serotonergic-calcium axis. Specifically, fluoxetine increases serotonin reuptake by the mammary gland, stimulating downstream signaling cascades responsible for bone resorption. Bone loss during lactation with use of SSRI may be too extensive for women to recover and be mediated by administration of folic acid.


Nothing to Disclose: SW, LLH, CV

OR09-6 25971 6.0000 A Use of Selective Serotonin Reuptake Inhibitors Promotes Bone Resorption during Lactation 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Reproductive Endocrinology Friday, April 1st 1:15:00 PM OR09 7767 11:45:00 AM Role of Growth Factor Signaling in Fertility and Pregnancy Oral

Bin Xu*1, Jeffrey O'Donnell2, Michael O'Donnell3, Jingcheng Yu4 and Ronald J Koenig5
1University of MIchigan, Ann Arbor, MI, 2University of MIchigan, 3University of Michigan, 4University of Michigan, Ann Arbor, MI, 5University of Michigan Medical Center, Ann Arbor, MI


Approximately 35% of follicular thyroid carcinomas and a small fraction of follicular adenomas are associated with a t(2;3)(q13;p25) chromosomal translocation that fuses paired box gene 8 (PAX8) with the peroxisome proliferator-activated receptor-gamma gene (PPARγ), resulting in expression of a PAX8-PPARγ fusion protein, PPFP. We previously generated a transgenic mouse model that combines Cre-dependent PPFP expression (PPFP;Cre) with homozygous deletion of floxed Pten (PtenFF;Cre), both thyroid specific since Cre expression is driven by the TPO promoter. We have shown that the combined PPFP;PtenFF;Cre mice develop metastatic thyroid cancer, but feeding the PPARγ agonist pioglitazone decreased the size of the primary tumor and prevented metastatic disease in vivo. The antitumor effect correlates with the fact that pioglitazone turns PPFP into a strongly PPARγ-like molecule, resulting in trans-differentiation of the thyroid cancer cells into adipocyte-like cells that lose malignant character as they become more differentiated. To further study this process, we created cell lines from the PPFP;PtenFF;Cre mouse thyroids and characterized the role of pioglitazone in their trans-differentiation into adipocyte-like cells in vitro. Our data show that pioglitazone induced cellular lipid accumulation and the expression of adipocyte marker genes such as FABP4, Plin1, CD36 and LPL. Knockdown of PPFP by shRNA eliminates the effect of pioglitazone on promoting adipogenic differentiaion with significant down regulation of a subset of adipocyte marker genes. The thyroid transcription factor 1 (TTF-1; NKX2-1) is highly expressed in thyroid cells, and is a critical transcription factor that regulates thyroid specific gene expressions.  We have found that PPFP and TTF-1 physically interact. To study the functional significance of this interaction, we used shRNA to silence TTF-1 expression in rat PCCL3 thyroid cells that stably express PPFP. We found that TTF-1 knockdown enhanced the adipogenic differentiation of these cells by pioglitazone, as assessed both by lipid content and expression of adipogenic genes. Silencing of TTF-1 was found to up regulate a broad subset of PPFP target genes in a PPFP- and pioglitazone-dependent manner. Our data suggest that the ability of pioglitazone/PPFP to promote thyrocyte/cancer cell adipogenic differentiation may be inhibited by TTF-1.


Nothing to Disclose: BX, JO, MO, JY, RJK

OR11-1 24619 1.0000 A Adipogenic Differentiation of Thyroid Cancer Cells through the Pax8-Pparg Fusion Protein Is Regulated By Thyroid Transcription Factor 1 (TTF-1) 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Thyroid/HPT Axis Friday, April 1st 1:15:00 PM OR11 7774 11:45:00 AM Thyroid Hormone Action, Cancer and Clinical Thyroid Oral

An Yang*1, Kaitlyn Liu2, Laura Perin3, Sheue-Yann Cheng4, Gregory A Brent5 and Anna Milanesi1
1Department of Medicine, Veteran Affair Greater Los Angeles Healthcare System and Department of Medicine and Physiology, David Geffen School of Medicine at UCLA, Los Angeles, CA, 2David Geffen School of Medicine at UCLA and Department of Medicine, Veteran Affair Greater Los Angeles Healthcare System, Los Angeles, CA, 3Department of Urology, Children’s Hospital Los Angeles, Los Angeles, CA, 4NIH - Nat'l Cancer Inst, Bethesda, MD, 5Veteran Affair Greater Los Angeles Healthcare System and David Geffen School of Medicine at UCLA, Los Angeles, CA


Myopathic changes, including muscular atrophy and weakness, are commonly described in hypothyroid and hyperthyroid patients. The genomic actions of triiodothyronine (T3) are mediated by thyroid hormone nuclear receptors (TRs) which are ligand-inducible transcription factors found in all tissues.   Myofiber-associated mononuclear satellite cells (SCs) are stem cells located in close proximity to muscle fibers beneath the basal lamina and play a major role in skeletal muscle maintenance and regeneration after injury. The self-renewing and proliferation of SCs lead to new myofiber formation and is crucial for the maintenance of the SC pool.  Paired box transcription factor 7 (Pax7) coordinates  self-renewal and maintenance of the SC niche and is required to instruct the myogenic fate of SCs. We recently reported impaired skeletal muscle regeneration in a mouse model of Resistance to Thyroid Hormone (RTH)-TRα1PV (a frame-shift mutation)1. One salient finding was a significant reduction of PAX7-positive SCs in the skeletal muscle of TRα1PV mice, suggesting that the exhaustion of the SC pool was the primary reason for the impaired skeletal muscle regeneration.      

To test this hypothesis, we analyzed the skeletal muscle of TRα1PV mice 4 days after cardiotoxin-induced muscle injury and compared with wild type (WT) animals. We injected TRα-knockdown C2C12 myoblasts into injured skeletal muscle and compared the in vivo behavior, 4 days after transplantation, with control C2C12 myoblasts. The cells were labeled with DiI to facilitate in vivo tracking.   

We found significant muscle inflammation in TRα1PV mice, 4 days after muscle injury, compared with WT mice.  The total number of PAX7-positive SCs in skeletal muscle of TRα1PV mice was significantly lower than control, both before and 4 days after muscle injury, indicating SC pool exhaustion. PAX7/Ki67 double staining after muscle injury showed significantly lower SC proliferation in TRα1PV mice. Ki67 was co-expressed in about 50% of the PAX7-expressing SCs in the control muscle, but less than 5% in the TRα1PV muscle. The mice injected with control-SCs showed efficient engraftment and migration of the donor cells in close proximity to the muscle fiber. Mice injected with TRα-knockdown SCs showed few donor cells localized in the sub laminar space.

Our data indicate that TRα plays an important role in maintenance of the SC niche. Impaired skeletal muscle regeneration in TRα1PV mice may be explained by insufficient SC activation and proliferation, and progressive loss of the satellite cell pool. Regulation of SC proliferation provide a therapeutic target  to enhance skeletal muscle regeneration.


Nothing to Disclose: AY, KL, LP, SYC, GAB, AM

OR11-2 26073 2.0000 A Thyroid Hormone Receptor Alpha Is Important for Maintenance of Skeletal Muscle Satellite Cell Niche In Vivo 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Thyroid/HPT Axis Friday, April 1st 1:15:00 PM OR11 7774 11:45:00 AM Thyroid Hormone Action, Cancer and Clinical Thyroid Oral

Sun Wook Cho*1, Young Shin Shong2, Hyun Jin Sun3, Do Joon Park2, Ka Hee Yi2 and Young Joo Park2
1Seoul National University College of Medicine, Seoul, 2Seoul National University College of Medicine, Seoul, Korea, Republic of (South), 3Seoul National University Hospital, Seoul, Korea, Republic of (South)


Thyroid stimulating hormone (TSH) suppression is a well-established prognostic factor for papillary thyroid cancer (PTC). However, the mechanism of TSH action on the initiation and progression of PTC remains elusive. The aim of this study was to investigate the role of TSH on PTC tumor growth, focusing on tumor microenvironment.

Tumorigenic clone of PTC cells (BHP10-3SCp) were implanted in nude mice, following recombinant human TSH (rhTSH) or saline for 3 weeks (n=10/each). Tumors of rhTSH group were bigger than saline group from day15 to day 20 (1733.4±793.5 vs 1148.8±471.1mm3, p<0.05). At day 20, 3 of 10 mice showed tumor bleeding in rhTSH group, while saline group showed none. Immunofluorescent staining of CD31 showed higher density of vasculature in rhTSH and saline group. Intravascular injection of FITC-labeled nanoparticles showed dilated tortious vascularity in rhTSH group compared to saline group. Moreover, tumors of rhTSH group showed higher density of F4/80-positive macrophages than that of saline group. To evaluate the mechanistic insight of the TSH actions on tumor angiogenesis, TSH were treated to BHP10-3SCp cells and vascular endothelial growth factor (VEGF) expressions were evaluated. mRNA expressions were up-regulated by 3-fold and 9-fold at 48 and 72 hrs, respectively. Conditioned medium (CM) of TSH-treated BHP10-3SCp cells showed increased VEGF concentrations than that of control-CM. Next, TSH-CMs were treated into microvascular endothelial cells (HMVEC) or macrophages (THP-1). TSH-CM treated group showed enhanced HMVEC migration and tube formation potentials than control-CM group, and these effects were attenuated with treatment of Bevacizumab, a humanized anti-VEGF antibody. Finally, TSH-CM also increased migration potentials in THP-1 cells.

In conclusion, TSH supported PTC tumor growth by enhancing tumor angiogenesis and macrophage recruitment into tumor microenvironment and its action on PTC tumor angiogenesis was partly mediated by VEGF, which may be a potential therapeutic target in TSH-dependent PTC progressions.


Nothing to Disclose: SWC, YSS, HJS, DJP, KHY, YJP

OR11-3 27535 3.0000 A Thyroid Stimulating Hormone Promotes Tumor Growth By Modulating Angiogenesis and Macrophage Recruitment in Papillary Thyroid Cancer Microenvironment 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Thyroid/HPT Axis Friday, April 1st 1:15:00 PM OR11 7774 11:45:00 AM Thyroid Hormone Action, Cancer and Clinical Thyroid Oral

Jeongwon Park*, Won Gu Kim, Li Zhao, Keisuke Enomoto, Mark Willingham and Sheue-Yann Cheng
NIH - Nat'l Cancer Inst, Bethesda, MD


Compelling epidemiologic evidence indicates that obesity is associated with a high risk of human malignancies, including thyroid cancer. In recent decades, the incidence of thyroid cancer has dramatically increased as have obesity rates. Large population studies further suggest that high body mass index is closely linked to the risk of advanced thyroid cancer. However, the underlying molecular mechanisms by which obesity could contribute to thyroid cancer progression remain unclear. We previously demonstrated that a high fat diet (HFD) effectively induced the obese phenotype in a mouse model of aggressive follicular thyroid cancer (ThrbPV/PVPten+/- mice). We showed that HFD promotes cancer progression through aberrant activation of the leptin-JAK2-STAT3 signal pathway. HFD promotion of thyroid cancer progression allowed us to test other molecular targets for therapeutic opportunity for obesity-induced thyroid cancer. Metformin is a widely used drug to treat patients with type 2 diabetes, especially in over weight patients. Metformin has been shown to reduce incidences of neoplastic diseases and cancer mortality in 2 diabetes patients. Our study aimed to test whether metformin could be a therapeutic for obesity-induced thyroid cancer.  Accordingly, ThrbPV/PVPten+/- mice were treated with HFD together with metformin or vehicle-only as controls for 20 weeks. While ThrbPV/PVPten+/- mice fed with HFD decreased survival as compared with vehicle-treated mice, metformin had no effects on the survival of HFD-treated ThrbPV/PVPten+/- mice.  Thyroid tumor growth of HFD-treated ThrbPV/PVPten+/- mice trended lower by metformin treatment. The apparent decreased tumor growth was due to increased apoptosis as evidenced by greater nuclear cleaved caspase 3 activity and suppression of the mTOR downstream p70S6K and 4E-BP1 protein abundance to decrease protein synthesis.  Remarkably, metformin markedly decreased occurrence of capsular invasion and completely blocked vascular invasion and anaplasia in HFD-treated ThrbPV/PVPten+/- mice.  The impeded cancer progression was due to the inhibitory effect of metformin on STAT3-vimentin and ERK signaling to decrease tumor cell invasion and de-differentiation.  The present studies have provided additional molecular evidence to support the link of obesity to thyroid cancer risks. Importantly, our findings suggest that metformin could be a novel treatment strategy for obesity-induced thyroid cancer.


Nothing to Disclose: JP, WGK, LZ, KE, MW, SYC

OR11-4 25278 4.0000 A Metformin Blocks Progression of Obesity-Induced Thyroid Cancer in a Mouse Model 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Thyroid/HPT Axis Friday, April 1st 1:15:00 PM OR11 7774 11:45:00 AM Thyroid Hormone Action, Cancer and Clinical Thyroid Oral

Meng Zhao*, Lu Liu, Qingbo Guan, Haiqing Zhang and Jiajun Zhao
Shandong Provincial Hospital affiliated to Shandong University, Jinan, China


Background Dyslipidemia is a serious public health problem for its disastrous effects. Subclinical hypothyroidism (SCH), one of the risk factors of dyslipidemia, has attracted the public attention recently. At least 75% of the SCH patients are diagnosed as mild SCH (thyrotropin≤10 mIU/L). However, it is still controversial in the management of mild SCH. It remains a debatable problem whether mild SCH patients could benefit from L-thyroxine replacement therapy on serum lipid profiles.

Methods This open-label, randomized, controlled trial was performed in Ningyang County, Shandong Province, China. Mild SCH patients who were diagnosed with twice thyroid function tests were randomly assigned to receive L-thyroxine replacement therapy (25 μg once daily initially, adjusted according to the reevaluation results until thyroid function returned normal) or no treatment for 15 months. The primary outcome was the change in serum total cholesterol (TC) concentrations analyzed in the per-protocol population. Subgroup analyses were also performed in subjects with different thyrotropin or TC concentrations at baseline. This study is registered with ClinicalTrials.gov, number NCT01848171.

Results Between July and December 2013, 378 subjects were enrolled and 369 subjects were included in the final analysis (210 in the intervention group and 159 in the control group). With the L-thyroxine replacement therapy, serum TC levels were decreased by 0.41 mmol/L (p<0.001) in the intervention group, while the decline was 0.17 mmol/L (p=0.019) in the control group. The decrease was more obvious in the intervention group than the control group (p=0.012). Subgroup analyses showed that in the subjects with different thyrotropin concentrations at baseline, L-thyroxine resulted in similar decreases of TC levels in each subgroup (all declines were approximately 0.41 mmol/L, p<0.001). In addition, L-thyroxine replacement therapy could also benefit the subjects with different TC concentrations at baseline. Even in the subjects with absolutely normal TC levels (less than 5.18 mmol/L), serum TC concentrations were kept unchanged in the intervention group (p=0.936), while they were increased by 0.35 mmol/Lin the control group (p=0.004). Variation trend for serum low-density lipoprotein cholesterol levels was similar with that for serum TC levels.

Conclusion Mild SCH patients could benefit from L-thyroxine replacement therapy on serum lipid profiles. Our study might provide reliable and important evidence for evidence-based medicine and help clinicians offer effective treatment advices to the mild SCH patients.


Nothing to Disclose: MZ, LL, QG, HZ, JZ

OR11-5 25826 5.0000 A Effects of L-Thyroxine Replacement Therapy on Serum Lipid Profiles in Patients with Mild Subclinical Hypothyroidism: An Open-Label, Randomized, Controlled Trial 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Thyroid/HPT Axis Friday, April 1st 1:15:00 PM OR11 7774 11:45:00 AM Thyroid Hormone Action, Cancer and Clinical Thyroid Oral

Paige Meizlik*1, Alice Arnold2, Michelle Carlson3, John Robbins4, Bruce Psaty2 and Anne R Cappola5
1Hospital of the University of Pennsylvania, Philadelphia, PA, 2University of Washington, Seattle, WA, 3Johns Hopkins University, Baltimore, MD, 4UC Davis Health System, Sacramento, CA, 5University of Pennsylvania, Philadelphia, PA


Background:Thyroid hormone use is common in older people, and overreplacement occurs frequently. Endogenous subclinical hyperthyroidism is associated with increased risk of fracture and atrial fibrillation. However, at similar TSH concentrations, T4 levels are higher and T3 levels are lower in levothyroxine users, and extrapolation of findings from untreated endogenous thyroid dysfunction to thyroid hormone use may not be appropriate. We sought to determine the relationship between thyroid function and adverse outcomes in older adults taking levothyroxine.

Methods:We examined associations between TSH (mIU/L), free T4 (pmol/L), and total T3 (nmol/L) and incidence of hip fracture, atrial fibrillation, dementia, coronary heart disease (CHD), heart failure, and total mortality in 360 US community-dwelling men and women aged 65 years and over who were enrolled in the Cardiovascular Health Study and taking levothyroxine. Cox proportional hazard models were used to examine the relationship between each thyroid test and incident events; TSH was log-transformed. All models were adjusted for age, sex, race, and outcome specific covariates.

Results:Mean age was 74.8 years and 79% were women. Only 51% of these levothyroxine users were euthyroid; 13% had overt hyperthyroidism, 16% had subclinical hyperthyroidism, and 20% had an elevated TSH.  There were 28 cases of incident hip fracture. Higher TSH levels were associated with a lower risk of hip fracture (HR 0.84; 95% CI 0.71-0.98, p=0.03 for lnTSH) and, concordantly, higher free T4 levels were associated with a higher risk of hip fracture (HR 2.63, 95% CI 1.26-5.50, p=0.01). There were 31 cases of incident dementia and 57 cases of incident CHD. Higher free T4 levels were associated with a higher risk of dementia (HR 2.57, 95% CI 1.18-5.56), p=0.02) and a lower risk of CHD (HR 0.42, 95% CI 0.19-0.93, p=0.03). There were 57 cases of incident atrial fibrillation and 70 cases of incident heart failure. There were no associations between any thyroid function test and atrial fibrillation or heart failure. There were 122 deaths. Higher total T3 levels were associated with lower risk of mortality (HR 0.35, 95% CI 0.19-0.64, p=0.001), consistent with nonthyroidal illness syndrome.

Conclusions: Higher free T4 concentrations are associated with a higher risk of hip fracture and dementia, and a lower risk of CHD, in older levothyroxine users. Clinical trials are needed to evaluate the optimal target concentrations for thyroid hormone replacement in older people.


Nothing to Disclose: PM, AA, MC, JR, BP, ARC

OR11-6 24968 6.0000 A Thyroid Function Testing in Older Adults Taking Levothyroxine and Risk of Adverse Outcomes 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Thyroid/HPT Axis Friday, April 1st 1:15:00 PM OR11 7774 11:45:00 AM Thyroid Hormone Action, Cancer and Clinical Thyroid Oral

Carmela Maniero*1, Junhua Zhou1, Elena AB Azizan2, Giles S H Yeo3, Brian Y H Lam3, Sudeshna G Neogi4, Wanfeng Zhao5 and Morris J Brown1
1University of Cambridge, Cambridge, United Kingdom, 2The National University of Malaysia (UKM) Medical Centre, Kuala Lumpur, Malaysia, 3University of Cambridge Metabolic Research Laboratories, Cambridge, United Kingdom, 4Genomics CoreLab, Cambridge National Institute for Health Research (NIHR) Biomedical Research Centre (BRC), Cambridge, United Kingdom, 5Cambridge University Hospitals NHS Foundation Trust, Cambridge, United Kingdom


Objective: The commonest curable cause of hypertension, aldosterone producing adenomas (APAs) vary in their clinical presentation, and underlying molecular and pathological phenotype. We have investigated the molecular basis of variable aldosterone response to metoclopramide, a dopamine receptor 2 antagonist used to increase the sensitivity of adrenal vein sampling in diagnosing lateralization of aldosterone overproduction.

Design and Method: We compared the transcriptome of zona glomerulosa (ZG), zona fasciculata (ZF), and APAs in 13 human adrenals, finding 28 genes >5-fold over-expressed in ZG vs ZF1. Expression of neurofilament medium (NEFM), a regulator of dopamine receptors, was 14.8-fold higher (p= 9.16E-12) in ZG than ZF, and 4-fold down-regulated in ZF-like APAs carrying KCNJ5 mutations in comparison to small ZG-like APAs (p=-4.35E-03). NEFM protein expression and subcellular localisation were evaluated by immunohistochemistry of human adrenals, and immunofluorescence microscopy in H295R and HEK293 cells. Aldosterone production and secretion and the dopamine receptors gene expression were investigated by silencing NEFM in adrenocarcinoma H295R cells. Aldosterone response to dopamine receptor 2 antagonist metoclopramide (10-7M) was measured in H295R cells transfected with mutant DelI157 KCNJ5 constructs.

Results: ZG selective expression of NEFM was confirmed by qPCR at 200-fold (cf. ZF). Immunohistochemistry showed ZG selectivity for NEFM staining, which was cytoplasmic and peri-membranous in normal ZG and small ZG-like APAs but absent in the normal ZF and ZF-like APAs. NEFM fluorescence staining in H295R was not filamentous (as in neurons and in HEK293) but present in nuclei and cytoplasm. Silencing of NEFM in H295R cells caused significant down-regulation of CYP11B2 and NR4A2by 40% and 70%, respectively (p= 0.01 and 0.05), and 40% increase  of aldosterone secretion after 72 hrs (from 205 to 280 pM/ug protein, p= 0.004). Moreover, dopamine receptor 1 and 2 were down-regulated by 45% and 70%, respectively (p= 0.007 and 0.02).

Aldosterone increase in response to dopamine receptor 2 blockade with metoclopramide was blunted in H295R cells transfected with KCNJ5 mutant construct whereas it increased by 200% in controls (p= 0.0159).

Conclusions: We postulate that NEFM plays a role in regulating basal aldosterone production and its response to dopamine. Down-regulation of NEFM in ZF-like APAs could explain the low expression levels of dopamine receptor 2 observed in some APAs, and the variable responses of aldosterone secretion to metoclopramide.2


Nothing to Disclose: CM, JZ, EAA, GSHY, BYHL, SGN, WZ, MJB

PP02-1 27402 1.0000 FRI 572 A Nefm Is a Zona Glomerulosa Selective Gene Involved in Regulating Aldosterone Secretion Response to Dopamine 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 3:15:00 PM FRI 572-594 7662 1:15:00 PM Molecular Regulation and Diagnosis of Hyepraldosteronism (posters) Poster

Yoshiyu Takeda*1, Shigehiro Karashima2, Mitsuhiro Kometani2, Yoshimichi Takeda2, Masashi Demura2, Yusuke Wakabayashi2 and Takashi Yoneda2
1Kanazawa University, Ishikawa, Japan, 2Kanazawa University, Kanazawa, Japan


PurposeFWe found the hypomethylation status of CYP11B2 gene in the aldosterone-producing adenomas compared with normal adrenal glands or non-functioning adrenal adenomas and reported that the methylation of promoter region of this gene influenced the mRNA expression in the adrenal cells. Angiotensin II or potassium influenced the methylation status of CYP11B2 gene. In order to clarify the effect of salt intake on the methylation status of CYP11B2 gene, rats were fed with high or low salt diet and the methylation status of CYP11B2 gene in the adrenal gland was examined.     

MethodsFFive Wister rats were fed with high, normal and low salt diet for 8 weeks. Plasma and urinary aldosterone, PRA were measured by RIA. The gene expression of CYP11B2 was measured by real time quantitative PCR in the adrenal glands. Isolated DNAs from rat adrenal glands were treated with bisulfite and amplified using primers specific for the human CYP11B2 promoter regions.

ResultsFPlasma and urinary aldosterone and PRA were significantly decreased by high salt diet compared with normal or low salt diet (p<0.05). High salt diet significantly decreased mRNA levels of CYP11B2 gene and increased metylation ratio of this gene (p<0.05).

ConclusionsFHigh salt diet may influence the methylation ratio of CYP11B2 gene and regulate aldosterone biosynthesis.


Nothing to Disclose: YT, SK, MK, YT, MD, YW, TY

26457 5.0000 FRI 576 A Epigenetic Control of CYP11B2 Gene By High Salt Diet 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 3:15:00 PM FRI 572-594 7662 1:15:00 PM Molecular Regulation and Diagnosis of Hyepraldosteronism (posters) Poster

Giacomo Rossitto*1, Diego Miotto2, Michele Battistel2, Giulio Barbiero2, Valeria Bisogni1, Giuseppe Maiolino1 and Gian Paolo Rossi3
1Padua University, Hypertension Unit, Italy, 2Padua University, Institute of Radiology, Italy, 3Padue University - Hypertension Unit, Padova, Italy


Objective: As metoclopramide exerts a secretagogue effect on aldosterone, we investigated its effect on the relative aldosterone secretion index [RAI: (Aside/Cside)/(AIVC/CIVC)] of each adrenal gland and its impact on the diagnostic accuracy of adrenal vein sampling (AVS) in primary aldosteronism (PA).

Method: in a within-patient design, we compared baseline and post-metoclopramide values of lateralization index (LI) and relative aldosterone secretion index (RAI) of  each adrenal gland in 103 consecutive patients undergoing AVS, using a conclusive diagnosis of aldosterone-producing adenoma (APA) as gold standard.

Results: Metoclopramide increased aldosterone in inferior vena cava (IVC) and in adrenal vein blood of both APA and the contralateral side. Post-metoclopramide LI provided an accurate identification of APA (p < 0.001), but did not improve the diagnostic accuracy over baseline LI, because of a similar increase of RAI of both sides (p<0.005). The baseline non-dominant RAI  showed low accuracy for the identification of a contralateral APA. By contrast, the post metoclopramide RAI values raised consistently >1.00 on the APA side, and bilaterally in all but one (non-dominat RAI = 0.91) non-APA patients; at variance, it failed to do so in the side contralateral to the tumor in 46% of the APA patients.

Conclusions: albeit increasing aldosterone secretion, metoclopramide did not improve the assessment of lateralization. However, a post-metoclopramide RAI < 0.91 offered 46% sensitivity for identifying an APA contralaterally and 100% specificity. Thus post metoclopramide AVS can allow a diagnosis in many cases which would have been non-diagnosed because of non bilaterally-selective AVS studies.


Nothing to Disclose: GR, DM, MB, GB, VB, GM, GPR

26601 6.0000 FRI 577 A Effect of Metoclopramide on  Adrenal Vein Aldosterone and Cortisol Levels:  Implications for the Subtyping of Primary Aldosteronism 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 3:15:00 PM FRI 572-594 7662 1:15:00 PM Molecular Regulation and Diagnosis of Hyepraldosteronism (posters) Poster

Mitsuhiro Kometani1, Takashi Yoneda*1, Mikiya Usukura2, Shigehiro Karashima1 and Yoshiyu Takeda3
1Kanazawa University, Kanazawa, Japan, 2Kanazawa University, KANAZAWA, Japan, 3Kanazawa Univ Sch of Med, Ishikawa, Japan


Background: Adrenal venous sampling (AVS) is the most reliable procedure to subtype primary aldosteronism (PA). However, AVS procedures, especially for use of adrenocorticotropic hormone (ACTH) stimulation and the diagnostic indices vary between centers.

Objective: We evaluated diagnostic discrepancy in subtyping of PA by different criteria based on the PA guideline of both Endocrine Society (ENDO) and Japan Endocrine Society (JES).

Methods: We evaluated 105 consecutive PA patients undergoing AVS from 2011 to 2014. The selectivity index (SI) was PCCadrenal/PCCIVC, AVS was considered bilaterally successful when the SI on both sides was >3 without ACTH stimulation, and >10 with ACTH stimulation, respectively. Lateralization index (LI) and contralateral index (CI) was calculated as PACdominant/PCCdominant/PACnon-dominant/PCCnon-dominant and PACnondominant/PCCnondominant /PACIVC/PCCIVC.  We used several criteria for unilateral PA as follows: 1st criteria was LI>2 and 2nd criteria was PACdominant>14000 pg/mL and 3rd criteria was LI >2.6 and 4th criteria was LI >4 after ACTH stimulation.

Results: Seventy-five out of 105(71%) cases were successful in baseline AVS and ACTH stimulated AVS at the same time. In baseline AVS, the prevalence of unilateral PA in 1st criteria were 38/75 (51%). Moreover, in ACTH stimulated AVS, the unilateral PA prevalence in 2nd, 3rd and 4th criteria were 28/75 (37%), 15/75 (20%) and 6/75 (8%), respectively. In every criteria, diagnostic discrepancy between baseline and ACTH stimulated AVS were observed in approximately 16-45% cases. In most of cases demonstrating diagnostic discrepancy, diagnoses were changed from unilateral to bilateral PA by ACTH injection. Furthermore, ACTH stimulation significantly decreased LI in all unilateral PA diagnosed on 1st criteria (p<0.05).

Conclusion: The criteria changed the diagnosis of PA subtype. Especially, ACTH stimulation increased the prevalence of bilateral PA. Further studies are necessary to clarify which AVS procedure is more accurate.

Abbreviations; PCC: plasma cortisol concertation, PAC: plasma aldosterone concertation, IVC: inferior vena cava


Nothing to Disclose: MK, TY, MU, SK, YT

25240 7.0000 FRI 578 A Diagnostic Discrepancy Between Baseline and ACTH Stimulated Adrenal Venous Sampling for Primary Aldosteronism 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 3:15:00 PM FRI 572-594 7662 1:15:00 PM Molecular Regulation and Diagnosis of Hyepraldosteronism (posters) Poster

Kohei Kamemura*1, Norio Wada2, Takamasa Ichijo3, Yuichi Matsuda4, Yuichi Fujii5, Tatsuya Kai6, Tomikazu Fukuoka7, Ryuichi Sakamoto8, Atsushi Ogo8, Tomoko Suzuki9, Hironobu Umakoshi10, Kazutaka Nanba10, Mika Tsuiki10 and Mitsuhide Naruse10
1Akashi Medical Center, Akashi, Japan, 2Sapporo City General Hospital, Sapporo, Japan, 3Saiseikai Yokohamashi Tobu Hospital, Yokohama, Japan, 4Sanda City Hospital, Sanda, Japan, 5Hiroshima General Hospital of West Japan Railway Company, Hiroshima, Japan, 6Saiseikai Tondabayashi Hospital, Japan, 7Matsuyama Red Cross Hospital, Matsuyama, Japan, 8Kyushu Medical Center, Fukuoka, Japan, 9Kitasato University, 10National Hospital Organization Kyoto Medical Center, Kyoto, Japan


Backgrounds: Although adrenal CT scan is the initial method to classify the subtypes in patients with primary aldosteronism (PA), laterality by the CT is not always concordant with that by adrenal vein sampling (AVS). AVS is however an invasive and a difficult procedure and whether all the patients with a diagnosis of PA should undergo AVS remains to be established.

Objectives: To investigate the accuracy of adrenal CT in subtype diagnosis and to develop a prediction score for bilateral subtype in PA patients with no adrenal tumor on CT.

Methods: WAVES-J database of PA patients underwent AVS between 2006 and 2013 at nine referral centers in Japan were studied. Total 393 patients who had undergone successful AVS with cosyntropin stimulation were included in this study. Concordant rate between the adrenal CT and AVS was evaluated. Variables for bilateral subtype were identified by multivariate logistic regression analysis and a prediction score was constructed. Receiver operating characteristic (ROC) curve was used to evaluate the diagnostic ability of the score.

Results: Forty two % (165/393) of the patients showed adrenal tumor(s) (a unilateral tumor in 40%, bilateral tumors in 2%), while no tumor was detected in the rest of the patients. The subtype diagnosis by CT was concordant with that by AVS in 68% (269/393): 38% (68/156) of patients with a unilateral tumor, 56% (5/9) of patients with bilateral tumors and 89% (204/228) of patients without a tumor were concordant with the subtype diagnosis by the AVS. Multivariate logistic regression analysis in patients without a tumor revealed that female gender (β=1.476, p=0.005), plasma aldosterone concentration (pg/ml) to plasma renin activity ratio ≤550 (β=1.115, p=0.023) and serum potassium ≥3.8 mEq/l (β=1.970, p=0.001) were the independent predictors of bilateral subtype. The prediction score for bilateral subtype based on 3 variables, which each one point was attributed to, showed that a score of 3 points had 40% sensitivity and 96% specificity, and that of 2 points had 82% sensitivity and 63% specificity in ROC curve analysis.

Conclusions: This study demonstrated that concordant rate of the subtype diagnosis by adrenal CT imaging and AVS in PA patients were high in those without adrenal tumor but not in those with adrenal tumors. Our prediction score could be useful in selecting patients with bilateral lesions who don’t need to undergo AVS, especially if no tumor was detected on CT.


Nothing to Disclose: KK, NW, TI, YM, YF, TK, TF, RS, AO, TS, HU, KN, MT, MN

24225 8.0000 FRI 579 A Significance of Adrenal CT Scan in Predicting Laterality and Indicating Adrenal Vein Sampling in Primary Aldosteronism 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 3:15:00 PM FRI 572-594 7662 1:15:00 PM Molecular Regulation and Diagnosis of Hyepraldosteronism (posters) Poster

Yui Shibayama*1, Norio Wada1, Hironobu Umakoshi2, Takamasa Ichijo3, Yuichi Fujii4, Kohei Kamemura5, Tatsuya Kai6, Ryuichi Sakamoto7, Atsushi Ogo7, Yuichi Matsuda8, Tomikazu Fukuoka9, Mika Tsuiki2, Tomoko Suzuki10 and Mitsuhide Naruse11
1Sapporo City General Hospital, Sapporo, Japan, 2National Hospital Organization Kyoto Medical Center, Kyoto, Japan, 3Saiseikai Yokohamashi Tobu Hospital, Yokohama, Japan, 4Hiroshima General Hospital of West Japan Railway Company, Hiroshima, Japan, 5Akashi Medical Center, Akashi, Japan, 6Saiseikai Tondabayashi Hospital, Japan, 7Kyushu Medical Center, Fukuoka, Japan, 8Sanda City Hospital, Sanda, Japan, 9Matsuyama Red Cross Hospital, Matsuyama, Japan, 10Kitasato University, 11Kyoto Medical Center, Kyoto, Japan


In primary aldosteronism (PA), adrenal vein sampling (AVS) is recognized as the most reliable method of distinguishing unilateral from bilateral disease. If aldosterone/cortisol (A/C) ratio in one adrenal vein is lower than that in the inferior vena cava (IVC), the result of AVS is determined as unilateral disease of the other side. In some of the patients, however, A/C ratio is bilaterally lower than that in the IVC (bilateral aldosterone suppression; BAS) despite of successful AVS confirmed by selectivity index of cortisol. Lateralization of aldosterone production is uninterpretable in such cases. The aim of this study was to investigate clinical significance of BAS in the subtype diagnosis of PA. This study was conducted as a part of multi-center collaborative study on AVS in Japan (WAVES-J study). Data of 491 patients with PA (222 male, 269 female, mean age 54.3 ± 11.3 yrs.) from 9 centers were analysed. Cannulation was confirmed as successful in 304 patients by selectivity index more than 2 before ACTH stimulation. BAS was seen in 29/304 (9.5%) patients. PRA was significantly lower in patients with BAS (median: 0.2; 25th-75th percentiles: 0.1-0.3) than those without BAS (0.3, 0.1-1.5) (p<0.01). PAC, aldosterone renin ratio (ARR), and prevalence of adrenal tumor on CT did not differ between the patients with and without BAS. After ACTH stimulation, in 29 patients with BAS, cannulation was sufficiently confirmed in 24 patients. BAS was disappeared in 22/24 patients although it was newly observed in 5 patients without BAS. ACTH loading significantly reduced the prevalence of BAS in AVS (p<0.05). Ordinary we sampled blood from left adrenal vein, which is distal to the junction of the inferior phrenic vein. In 11 patients, we additionally sampled it from the junction of the inferior phrenic vein and left adrenal vein as a common trunk. Consequently BAS was lost in one patient. Of 29 patients with BAS, 9 patients were performed unilateral adrenal resection. Of 9 patients, 7 patients accomplished ARR<200. This study showed BAS occurred in unexpected cases in AVS, and the patients with BAS sometimes appeared to have a unilateral disease. ACTH stimulation as well as blood sampling in the common trunk rather than the central vein could be useful in reducing this unfavorable phenomenon for subtype diagnosis in PA.


Nothing to Disclose: YS, NW, HU, TI, YF, KK, TK, RS, AO, YM, TF, MT, TS, MN

24848 9.0000 FRI 580 A Prevalence of Bilateral Aldosterone Suppression in Adrenal Vein Sampling for Patients with Primary Aldosteronism: A Multi-Center Collaborative Study in Japan (WAVES-J study) 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 3:15:00 PM FRI 572-594 7662 1:15:00 PM Molecular Regulation and Diagnosis of Hyepraldosteronism (posters) Poster

Hironobu Umakoshi*1, Mitsuhide Naruse2, Norio Wada3, Takamasa Ichijo4, Kohei Kamemura5, Yuichi Matsuda6, Yuichi Fujii7, Tatsuya Kai8, Tomikazu Fukuoka9, Ryuichi Sakamoto10, Atsushi Ogo10, Tomoko Suzuki11, Kazutaka Nanba2 and Mika Tsuiki1
1National Hospital Organization Kyoto Medical Center, Kyoto, Japan, 2Kyoto Medical Center, Kyoto, Japan, 3Sapporo City General Hospital, Sapporo, Japan, 4Saiseikai Yokohamashi Tobu Hospital, Yokohama, Japan, 5Akashi Medical Center, Akashi, Japan, 6Sanda City Hospital, Sanda, Japan, 7Hiroshima General Hospital of West Japan Railway Company, Hiroshima, Japan, 8Saiseikai Tondabayashi Hospital, Japan, 9Matsuyama Red Cross Hospital, Matsuyama, Japan, 10Kyushu Medical Center, Fukuoka, Japan, 11Kitasato University


Background: Adrenal venous sampling (AVS) is considered to be the most reliable diagnostic procedure to lateralize aldosterone excess in primary aldosteronism (PA). However, normative criteria have not been established mainly because of a lack of data in non-PA hypertensive patients.

Objective: Aim of the study was to investigate aldosterone concentration and its gradient in the adrenal vein of non-PA hypertensive patients.

Design, Setting and Participants: This retrospective study was conducted as a multi-center collaborative study involving nine referral centers in Japan. We studied the results of cosyntropin-stimulated AVS in 41 hypertensive patients who showed negative results in 2 confirmatory tests, captopril challenge test and saline infusion test. AVS was conducted by the decision of respective attending physicians based on various clinical settings.

Main outcome measures: Plasma aldosterone concentration (PAC), aldosterone to cortisol (A/C) ratio, its higher to lower ratio (lateralized index: LI) in the adrenal vein with cosyntropin stimulation were measured.

Results: Median PAC in the adrenal vein was 25737 pg/ml (range: 5154-69920) in the higher side and 13347 pg/ml (range: 1866-36190) in the lower side (P<0.001). There was a significant gradient in A/C ratio between the higher and lower sides [27.2 pg/ml/µg/dl (5.4-66.0) vs 17.3 pg/ml/µg/dl (4.0-59.0); P<0.001] with LI ranging from 1.01 to 3.87. The aldosterone lateralization gradient was between 1 to 2 in 33 patients and 2 to 4 in 8 patients. None of the patients showed LI ≥4.

Conclusions: The present study demonstrated that PAC in the adrenal veins showed significant variation and lateralization gradient even in non-PA hypertensive patients. AVS aldosterone lateralization gradients between 2 and 4 should be interpreted with caution in patients with PA because these gradients can be found in patients without PA.


Nothing to Disclose: HU, MN, NW, TI, KK, YM, YF, TK, TF, RS, AO, TS, KN, MT

24686 10.0000 FRI 581 A Adrenal Venous Sampling in Patients without Primary Aldosteronism 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 3:15:00 PM FRI 572-594 7662 1:15:00 PM Molecular Regulation and Diagnosis of Hyepraldosteronism (posters) Poster

Yuto Yamazaki*1, Yasuhiro Nakamura2, Kazue Ise1, Kei Omata3, Yoshikiyo Ono3, Yuta Tezuka1, Ryo Morimoto3, Fumitoshi Satoh1 and Hironobu Sasano1
1Tohoku University Graduate School of Medicine, Sendai, Japan, 2Tohoku University Graduate School of Medicine, Sendai Miyagi, Japan, 3Tohoku University Hospital, Sendai, Japan


Primary aldosteronism (PA) is one of the main causes of secondary hypertension. The frequency of tumor undetected (CT negative) PA, including UAH (Unilateral Adrenal Hyperplasia), UMN (Unilateral adrenocortical micronodules) and IHA (Idiopathic hyperaldosteronism) constitutes approximately 20% of these cases. However, histological classification of these lesions has not necessarily been well-characterized. Recently, aldosterone producing cells can be readily identified by immunohistochemistry of CYP11B2, but the status of aldosterone biosynthesis of these CT negative PA has remained virtually unexplored. Therefore, in this study, we evaluated 15 cases clinically diagnosed as CT negative PA and analyzed the aldosterone biosynthesis using immunohistochemistry. Immunoreactivity of steroidogenic enzymes were evaluated by HSCORE. CYP11B2 (B2) immunohistochemistry revealed that 15 cases were tentatively classified into three histological diagnosis; 7 cases of unilateral adrenocortical micronodules (UMN), 5 cases of diffuse hyperplasia of ZG with multiple micronodules (DHMN) and 3 cases of micro aldosterone producing adenoma (micro APA) with aldosterone producing cell clusters (APCCs). UMN was defined as B2 positive micronodular hyperplasia with the adjacent ZGs negative for B2 and DHMN as the diffuse hyperplastic B2 positive ZG with micronodular hyperplasia also positive for B2. In addition, HSD3B1 immunoreactivity was siginificantly higher both in the area of hyperplastic nodules (P=0.04<0.05) and in the adjacent ZGs (P=0.0128<0.05) in UMN than in DHMN. HSD3B2 immunoreactivity was significantly higher in the area of the adjacent ZGs in DHMN than in UMN (P=0.0192<0.05). Results of this study revealed that hyperplastic lesion of CT negative PA can be classified into two groups according to the patterns of B2 immunolocalization; UMN (focal) and DHMN (diffuse). In addition, patterns of immunolocalization of HSD3B isoforms were different between these two histological subtypes. B2 immunostaining could also identify microAPA which cannot be discerned in routine histology. However, definitive differential diagnosis between microAPA with APCCs and UMN still requires further evaluation.


Nothing to Disclose: YY, YN, KI, KO, YO, YT, RM, FS, HS

24692 11.0000 FRI 582 A New Histological Classification of Primary Aldosteronism ~CYP11B2 Localization in CT Negative PA~ 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 3:15:00 PM FRI 572-594 7662 1:15:00 PM Molecular Regulation and Diagnosis of Hyepraldosteronism (posters) Poster

Tsutomu Abe*1, Mitsuhide Naruse2, William F Young Jr.3, Nobuya Kobashi1, Yoshihiro Doi1, Akihiro Izawa1, Kei Akama1, Yuki Okumura1, Miho Ikenaga1, Hiroyuki Kimura4, Hideo Saji4, Kuniaki Mukai5 and Hiroki Matsumoto1
1Nihon Medi-Physics Co. Ltd., Sodegaura, Japan, 2Kyoto Medical Center, Kyoto, Japan, 3Mayo Clinic, Rochester, MN, 4Kyoto University, Kyoto, Japan, 5School of Medicine, Keio University, Tokyo, Japan


Context: Primary aldosteronism (PA) is the most common and surgically curable form of endocrine hypertension. In patients with unilateral aldosterone hypersecretion, e.g., aldosterone-producing adenoma (APA), hypertension and hyperaldosteronism can be cured by unilateral adrenalectomy. Therefore, an accurate method for distinguishing unilateral from bilateral adrenal disease is critical. Adrenal vein sampling (AVS) is the standard method to distinguish unilateral from bilateral forms of PA. However, AVS is an invasive and technically difficult procedure. 11C-metomidate (MTO)-positron emission tomography (PET) was reported as a potential replacement for AVS. However, because MTO has low selectivity for CYP11B2 (aldosterone synthase) over CYP11B1 (11-beta-hydroxylase), dexamethasone pretreatment is essential for the imaging. In addition, PET imaging agents labeled with carbon-11 have limited utility because of the very short half-life of carbon-11. Therefore, longer half-life fluorine-18 or iodine-123-labeled imaging agents with a higher selectivity for CYP11B2 over CYP11B1 is desirable for APA imaging.

Objective: This study aimed to determine the selectivity of the new imaging agents 18F-CDP2230 (PET) and 123I-CDP2440 (single photon emission computed tomography: SPECT) for CYP11B2 over CYP11B1 and determine whether the biodistribution profiles are favorable for imaging CYP11B2.

Methods: The half maximal inhibitory concentrations (IC50) of CDP2230 and CDP2440 for the enzymatic activities of CYP11B2 and CYP11B1 were determined using cells with stable expression of either enzyme. In vitro autoradiography (ARG) of human adrenal sections with APA was performed to confirm the specific binding ability of the new imaging agents to CYP11B2-expressing regions. Biodistribution studies were performed in rats.

Results: The selectivity of CDP2230 and CDP2440 for CYP11B2 over CYP11B1 was higher than that of metomidate analogues. In vitro ARG revealed that the binding of the new imaging agents to CYP11B2-expressing regions in the adrenal gland were more specific than that of the metomidate analogue. Moreover, the biodistribution study showed that the new imaging agents accumulated in adrenal glands with low background uptake.

Conclusions: Our study revealed high selectivity of the new imaging agents for CYP11B2 over CYP11B1 with favorable biodistribution for imaging CYP11B2. 18F-CDP2230 and 123I-CDP2440 are promising imaging agents for the subtype diagnosis of PA.


Disclosure: TA: Employee, Nihon Medi-Physics Co. Ltd.. MN: Researcher, Research Center, Nihon Medi-Physics Co. Ltd.. WFY Jr.: Consultant, Research Center, Nihon Medi-Physics Co. Ltd.. NK: Employee, Nihon Medi-Physics Co. Ltd.. YD: Employee, Nihon Medi-Physics Co. Ltd.. AI: Employee, Nihon Medi-Physics Co. Ltd.. KA: Employee, Nihon Medi-Physics Co. Ltd.. YO: Employee, Nihon Medi-Physics Co. Ltd.. MI: Employee, Nihon Medi-Physics Co. Ltd.. HK: Researcher, Nihon Medi-Physics Co. Ltd.. HS: Researcher, Research Center, Nihon Medi-Physics Co. Ltd.. HM: Employee, Nihon Medi-Physics Co. Ltd.. Nothing to Disclose: KM

24419 12.0000 FRI 583 A A Novel CYP11B2-Specific Imaging Agent for Detection of Aldosterone-Producing Adenomas 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 3:15:00 PM FRI 572-594 7662 1:15:00 PM Molecular Regulation and Diagnosis of Hyepraldosteronism (posters) Poster

Takashi Yoneda*1, Masashi Demura2, Shigehiro Karashima2, Mitsuhiro Kometani2, Mikiya Usukura3, Masakazu Yamagishi2 and Yoshiyu Takeda1
1Kanazawa University, Ishikawa, Japan, 2Kanazawa University, Kanazawa, Japan, 3Kanazawa University, KANAZAWA, Japan


Background: Adrenal vein sampling (AVS) is essential for identifying a surgically curable form of primary aldosteronism (PA). However, the AVS success rate remains low, especially for non-ACTH stimulated AVS, because accurate placement of the sampling catheter is technically challenging. Intraprocedural cortisol measurement can confirm the catheter’s position, thereby increasing the AVS success rate.

Methods: We developed a novel quick cortisol assay (QCA) that uses immunochromatography and gold nanoparticles, and that can be performed either semi-quantitatively or quantitatively (United States Patent No. 8927219, Japan Patent No.5590651). The assay was evaluated during non-ACTH stimulated AVS in two prospective randomized, controlled studies. In a single-center study, PA patients were assigned to undergo AVS incorporating the semi-quantitative QCA (n=30), quantitative QCA (n=30), or without the QCA (n=30); and the rates of successful AVS were determined. The multicenter study was performed at seven medical centers, including Kanazawa University Hospital(K)(Kanazawa, Japan), Houju Memorial Hospital(H)(Nomi, Japan), Sanda Municipal Hospital(S)(Sanda, Japan), Akashi Medical Center(A)(Akashi, Japan), Ishikawa Prefectural Hospital(I)(Kanazawa, Japan), Saiseikai Ishikawa Hospital(SI)(Kanazawa, Japan), and Takaoka Municipal Hospital(T)(Takaoka, Japan) from March 2012 to March 2015. In a multicenter study, the success rates of AVS performed with (n=148) or without the semi-quantitative QCA (n=145) were determined.

Results: Cortisol concentrations were measured during AVS within 6 min in the radiology suite, without additional technical assistance, and excellently correlated with a conventional reference assay (r=0·997, p<0·001). In the single-center study, there was no difference in the AVS success rates using the semi-quantitative (93%) and quantitative QCAs (93%); however, both rates were significantly higher than the rate of AVS without QCA (63%; p<0·001). The success rate of AVS performed in the multicenter study was 94% for the semi-quantitative QCA, which was significantly higher than the rate for the control patients (56%; p<0·001).

Conclusions: Our novel QCA was rapidly and easily performed at the point-of-care, and improved the AVS success rate.


Disclosure: TY: Patent owner, Trust Medical C.O.. Nothing to Disclose: MD, SK, MK, MU, MY, YT

25736 13.0000 FRI 584 A Impact of New Quick Gold Nanoparticle-Based Cortisol Assay during Adrenal Vein Sampling for Identifying Surgically Curable Primary Aldosteronism 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 3:15:00 PM FRI 572-594 7662 1:15:00 PM Molecular Regulation and Diagnosis of Hyepraldosteronism (posters) Poster

Alejandra Tapia-Castillo*1, Jaime I Lizama2, Carolina P Valdivia2, Antonio Zapata2, Virginia Iturrieta2, Fidel Allende2, Sandra Solari2, Paula Villarzu2, Carmen Campino2, Alejandro Martínez-Aguayo2, Hernan Garcia2, Doris Muñoz2, Maria Paulina Rojas2, Andrea Vecchiola2, Gabriela Repetto1, Rene Baudrand2, Carlos E Fardella2 and Cristian A Carvajal2
1Universidad del Desarrollo, Santiago, Chile, 2Pontificia Universidad Catolica de Chile, Santiago, Chile


About 15% of essential hypertensives may suffer an impairment of the 11beta-hydroxysteroid dehydrogenase type 2 (11βHSD2) enzyme, which inactivates cortisol (F) to cortisone (E). However, the frequency of mutations in the HSD11B2 gene is extremely low to explain this prevalence. Thus, epigenetic mechanisms could explain the impairment seen in 11βHSD2 activity. Few studies reveal the role of miRNA on HSD11B2 expression. Tissue-specific exosomes containing miRNAs and proteins, may be a useful tool to get insight about pathophysiologic mechanisms involved in essential hypertension.

Aim: To identify, evaluate and compare the expression levels of miRNAs with potential binding sites on the 3'-UTR of the human HSD11B2 mRNA, in urinary exosomes and peripheral leukocytes  (PBMC).

Subjects and Methods: Subjects were classified in two groups according their serum F/E ratio: high F/E ratios (> percentil 90) and subjects with normal F/E ratio. Total RNA was isolated from PBMC (n=94) and in a subset spot-urinary exosomes (n=20). To identify exosomes we used electronic microscopy and aquaporin 2 (AQP2) expression. Bioinformatic analyses were performed by MirWalk 2.0, which identified potential miRNAs binding to HSD11B2 RNA. Expression of HSD11B2, miRNAs, AQP2 and RNU6 (housekeeping gene) were evaluated by qRT-PCR. The results were expressed as relative units (RU). Comparisons were performed by Kruskal-Wallis and Dunn analyses. 

Results: Bioinformatic analyses identified 12 miRNAs that potentially bind to the 3'UTR region of HSD11B2. We successfully amplified 4 miRNA in leukocytes (miR-101, 488, 615 and 1205) and 2 miRNA in exosomes by qRT-PCR, being miR-488 and miR-615 expressed in both samples. AQP2 expression was only expressed in urinary exosomes and not in PBMC. In the total group, the expression in PBMC of miR-1205 (8.9 [4.7 - 19.0] RU) was higher than expression of miR-101 (0.3 [0.1- 1.3] RU; p <0.05), miR-615 (2.1 [0.7- 5.1] RU; p <0.05) and miR-488 (0.4 [0.1- 1.3] RU; p <0.05). In the total group, we observed an increased expression of miR-615 and miR-488 in exosomes than PBMC (241.2 [27.6-4143] vs 2.1 [0.7-5.1] UR; p <0.0001) and (630.9 [91.2-1080] vs 0.4 [0.1-1.3] RU; p 0.0015), respectively. In urinary exosomes, the expression of miR-488 was about 6 times higher in subjects with high F/E ratios (n = 10) vs normal F/E ratios (n = 10) (1025 [915.1-1134] vs 176.4 [6.1 to 346 7] RU; p <0.0001), and the expression of HSD11B2 tended to be lower in the group subject to high F/E vs normal F/E (1150 [376-1968] vs 4730 [1312-8108] RU; p = 0.1). We found no differences in expression of miRNA in leukocytes.

Conclusion: We identified a high expression of miR-488 and miR-615 that could modify the expression of HSD11B2 when evaluated in urinary exosomes from subjects with high F/E ratio. We provide new evidence regarding the usefulness of exosomes and their content of miRNA as regulators and potential biomarkers of 11βHSD2 activity.


Nothing to Disclose: AT, JIL, CPV, AZ, VI, FA, SS, PV, CC, AM, HG, DM, MPR, AV, GR, RB, CEF, CAC

24594 14.0000 FRI 585 A Mir-488 and Mir-615 Identified in Urinary Exosome As Potential Modulators of HSD11B2 Expression 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 3:15:00 PM FRI 572-594 7662 1:15:00 PM Molecular Regulation and Diagnosis of Hyepraldosteronism (posters) Poster

Dai Suzuki*1, Akiko Saito-Hakoda2, Ryo Ito2, Kyoko Shimizu2, Naotaka Kogure2, Hiroyuki Kagechika3, Shigeo Kure1, Atsushi Yokoyama2 and Akira Sugawara2
1Tohoku Univ Grad Sch of Med, Sendai, Japan, 2Tohoku Univ Grad School of Med, Sendai, Japan, 3Tokyo Med Dent Univ


Background: Aldosterone secreted from adrenal gland is one of the most important hormones involved in the blood pressure regulation, and its increase contributes to the progression of resistant hypertension. The purpose of the study is to examine the effect of RXR agonist PA024 on adrenal aldosterone synthase gene (CYP11B2) expression as well as aldosterone secretion, and elucidate its molecular mechanisms for the future innovation of novel anti-hypertensive/anti-arteriosclerotic drugs.

Methods and Materials: Angiotensin (A) II and RXR pan-agonist PA024 were used as reagents. We added various concentrations of PA024 on H295R cells derived from human adrenocortical carcinoma. We thereafter analyzed CYP11B2 mRNA expression by quantitative real-time PCR, CYP11B2 promoter activity by luciferase assay, aldosterone secretion by enzyme immunoassay, and intracellular Ca2+ level by Calcium Kit-Fluo 4, respectively.

Results: PA024 dose-dependently suppressed AII-induced CYP11B2 mRNA expression and CYP11B2 promoter activity. Additionally, high-dose PA024 significantly decreased aldosterone secretion. The suppression of CYP11B2 promoter activity by PA024 was observed in the deletion mutants analyses using the region from -1521 (full length) to -135 including NBRE-1, Ad4, and Ad5 elements. Therefore, transcription factors binding to these elements may be involved in the suppression. PA024 also suppressed mRNA expression of transcription factors Nurr1 and NGFIB in a dose-dependent manner. Since PA024-mediated suppression of CYP11B2 promoter activity was rescued by Nurr1 overexpression, decrease of Nurr1 expression may contribute to the suppression. On the other hand, PA024 did not affect on intracellular Ca2+ concentrations, cellular proliferation, and apoptosis. PA024 also inhibited mRNA expression of StAR, HSD3β2, and CYP21A2. Therefore, PA024-mediated suppression of these enzymes (and protein) may also be involved in the inhibition of aldosterone synthesis pathway.

Conclusions: These data indicate that RXR pan-agonist PA024 may possibly be a candidate of novel anti-hypertensive/anti-arteriosclerotic drugs via the suppression of aldosterone synthesis/secretion. We are currently investigating its in vivo effect using transgenic Tsukuba hypertensive mice (hRN8-12 x hAG2-5) which demonstrate AII-mediated CYP11B2 overexpression.


Nothing to Disclose: DS, AS, RI, KS, NK, HK, SK, AY, AS

25208 15.0000 FRI 586 A Inhibitory Effects of Retinoid X Receptor Pan-Agonist PA024 on Adrenal CYP11B2 Expression and Aldosterone Secretion 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 3:15:00 PM FRI 572-594 7662 1:15:00 PM Molecular Regulation and Diagnosis of Hyepraldosteronism (posters) Poster

Hiroki Kobayashi*, Akira Haketa, Takahiro Ueno, Sho Tanaka, Yoshinari Hatanaka and Masayoshi Soma
Nihon University School of Medicine, Tokyo, Japan


Context: Currently, adrenal venous sampling (AVS) is the only reliable method to distinguish unilateral hyperaldosteronism (UHA) from bilateral hyperaldosteronism (BHA) in primary aldosteronism (PA). However, AVS is highly invasive and time consuming. Considering the limited availability of AVS compared with the high prevalence of PA, it would be helpful if we could gain additional information from routine clinical practice to select patients who should undergo AVS. The objective of this study is to compare the differences in circadian variation of serum adrenocortical hormone levels (i.e., aldosterone, cortisol, and ACTH) and 24-h urinary aldosterone level in UHA and BHA, and assess the diagnostic ability of plasma aldosterone concentration in different blood collecting times and other adrenocortical hormone levels for discriminating between UHA and BHA.

Setting and Design: This is a retrospective analysis of 64 patients who were diagnosed with PA and underwent AVS. Of these, 35 and 23 patients were diagnosed with UHA and BHA, respectively, including 25 patients pathologically confirmed as having APA. Their plasma adrenocortical hormone levels at 0:00, 6:00, 12:00, and 18:00 and 24-h urinary aldosterone level under the condition of 6 g daily dietary salt intake were measured.

Results: Serum sodium; PAC at 0:00, 6:00, 12:00 and 18:00; PFC at 12:00 and 18:00; and 24-h urinary aldosterone levels were statistically significantly higher in those with UHA than those with BHA. In contrast, the UHA group had lower levels of serum potassium and ACTH at 0:00. The area under the ROC curve for PAC at 0:00, 6:00, 12:00, and 18:00 and 24-h urinary aldosterone to discriminate UHA and BHA was 0.840 [95% confidence interval (CI); 0.73−0.95], 0.936 (95% CI; 0.87−1.00), 0.882 (95% CI; 0.79−0.98), 0.824 (95% CI; 0.71−0.94), 0.873 (95% CI; 0.78−0.97), respectively.

Conclusions: We firstly demonstrated the diagnostic value of PAC in different blood collecting times and 24-h urinary aldosterone level to discriminate between UHA and BHA. This test can be performed when we screen PA, and it is far less expensive than AVS. Thus, by omitting unnecessary AVS procedures, we can reduce the associated medical costs.


Nothing to Disclose: HK, AH, TU, ST, YH, MS

24079 16.0000 FRI 587 A Importance of Circadian Variation of Adrenocortical Hormone Levels and 24-h Urinary Aldosterone Concentration in the Subtype Evaluation of Primary Aldosteronism 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 3:15:00 PM FRI 572-594 7662 1:15:00 PM Molecular Regulation and Diagnosis of Hyepraldosteronism (posters) Poster

Hisae Ando*, Yoshinori Ozeki, Masahide Okamoto, Mitsuhiro Okamoto, Asami Sato, Yuichi Yoshida, Manabu Anai, So Ueda, Koro Gotoh, Takayuki Masaki, Tetsuya Kakuma and Hirotaka Shibata
Oita University, Yufu-city, Japan


Background: The widespread use of aldosterone to renin ratio results in increased prevalence of primary aldosteronism (PA).  To find surgically-curable cases, confirmatory tests which are followed by subtype diagnosis with adrenal venous sampling (AVS) are mandatory.  To investigate whether either confirmatory test can predict subtype diagnosis which was confirmed by AVS, we retrospectively investigated correlation between confirmatory test and AVS-confirmed laterality.

Methods: Seventy one hypertensive patients (male:female=33:38) were diagnosed as PA by confirmatory tests including captopril challenge, furosemide-upright posture, oral salt loading and rapid ACTH tests, and subtype diagnosis was performed by AVS (unilateral (UNI):bilateral  (BIL)=27:44).  The cut-off value for UNI was determined by ROC analysis and compared usefulness of confirmatory tests for subtype diagnosis.

Results: Blood pressure, pulse rate and BMI were comparable between UNI and BIL.  UNI PA patients present with younger age, lower serum potassium, higher plasma and urine aldosterone excretion.  Among 4 confirmatory test, rapid ACTH test was shown to be most useful to predict subtype diagnosis.  For UNI diagnosis, the cut-off value of peak aldosterone was 39.8ng/dL (83% sensitivity, 75% specificity, 59% positive predictive value, 91% negative predictive value).  The cut-off value of peak aldosterone/cortisol ratio was 1.63 (83% sensitivity, 71% specificity, 56% positive predictive value, 91% negative predictive value).

Conclusions: Rapid ACTH test may be an useful test for PA subtype diagnosis with a cut-off value of peak plasma aldosterone 39.8ng/dL or a cut-off value of peak plasma aldosterone/cortisol ratio 1.63.  When peak aldosterone or aldosterone/cortisol ratio is lower than cut-off values in rapid ACTH test, AVS can be skipped due to high probability of BIL PA.


Nothing to Disclose: HA, YO, MO, MO, AS, YY, MA, SU, KG, TM, TK, HS

25575 17.0000 FRI 588 A Usefulness of Rapid ACTH Test in Subtype Diagnosis of Primary Aldosteronism 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 3:15:00 PM FRI 572-594 7662 1:15:00 PM Molecular Regulation and Diagnosis of Hyepraldosteronism (posters) Poster

Xiaomu Li*1, Yan Ling2 and Xin Gao1
1Zhongshan Hospital, Fudan University, Shanghai, China, 2Department of Endocrinology, Zhongshan Hospital, Fudan University, Shanghai, China


Background:Long-term exposure to the high aldosterone levels may cause the renal damage in primary aldosteronism patients. Considering the possible associations of plasma renin activity (PRA) and plasma aldosterone concentration (PAC) with glomerular filtration rate (GFR), directly calculated aldosterone to renin ratio (ARR) levels, which is widely used as the screening criteria, may be significantly affected by GFR, especially in patients with long history of primary aldosteronism and renal damage. The aim of this study was to investigate the effect of estimated glomerular filtration (eGFR) to PRA, PAC and ARR levels in primary aldosteronism patients, and establish the diagnostic criteria of ARR by stratified eGFR.

Methods:One hundred and twenty-three patients with primary aldosteronism (PA) and one hundred and thirty-six patients with essential hypertension were consecutively recruited from Zhongshan hospital, Fudan university, 2012-2015. Statistical analysis was conducted with the Medcalc software.

Results:The results suggested that patients with primary aldosteronism had similar eGFR (P>0.05), higher PAC (P<0.001), lower PRA (P<0.001), and lower serum potassium levels (P<0.001), than patients with essential hypertension. eGFR was correlated inversely with PRA, and positively with calculated ARR, both in the patients with primary aldosteronism or essential hypertension. Primary aldosteronism patients from the lowest tertile of ARR (<17.8 pg/dl per ng/ml.h) or highest tertile of PRA (>1.4 ng/ml.h) were associated with higher BP, plasma potassium, and albuminuria and lower eGFR than other patients (P<0.01). In the whole study population, the optimal cut- off point according to Youden’s J statistic was an ARR of 25.2 pg/ml per ng/ml.h (AUC=0.845, 95% Confidence interval: 0.730 to 0.908, P<0.001); in subjects with eGFR higher than 90 ml/min/1.73m2, the optimal cut- off point was an ARR of 42.0 pg/dl per ng/ml.h (AUC=0.891, 95% Confidence interval: 0.789 to 0.972, P<0.001); in subjects with eGFR lower than 90 ml/min/1.73m2, the optimal cut- off point was an ARR of 19.3 (AUC=0.820, 95%CI: 0.714 to 0.911, P<0.001).

Conclusions: Unsuppressed renin and lower ARR levels were associated with more severe renal damage and decreased eGFR in patients with primary aldosteronism. Diagnostic criteria of ARR by stratified eGFR may be an optimal strategy for the screening of primary aldosteronism.


Nothing to Disclose: XL, YL, XG

24002 18.0000 FRI 589 A Effect of Estimated Glomerular Filtration Rate on Aldosterone to Renin Ratio in the Screening of Primary Aldosteronism 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 3:15:00 PM FRI 572-594 7662 1:15:00 PM Molecular Regulation and Diagnosis of Hyepraldosteronism (posters) Poster

Norio Wada*1, Yui Shibayama1, Hironobu Umakoshi2, Takamasa Ichijo3, Yuichi Fujii4, Kohei Kamemura5, Tatsuya Kai6, Ryuichi Sakamoto7, Atsushi Ogo7, Yuichi Matsuda8, Tomikazu Fukuoka9, Mika Tsuiki2, Tomoko Suzuki10 and Mitsuhide Naruse11
1Sapporo City General Hospital, Sapporo, Japan, 2National Hospital Organization Kyoto Medical Center, Kyoto, Japan, 3Saiseikai Yokohamashi Tobu Hospital, Yokohama, Japan, 4Hiroshima General Hospital of West Japan Railway Company, Hiroshima, Japan, 5Akashi Medical Center, Akashi, Japan, 6Saiseikai Tondabayashi Hospital, Japan, 7Kyushu Medical Center, Fukuoka, Japan, 8Sanda City Hospital, Sanda, Japan, 9Matsuyama Red Cross Hospital, Matsuyama, Japan, 10Kitasato University, 11Kyoto Medical Center, Kyoto, Japan


Hyperkalemia is an important complication of adrenalectomy for unilateral aldosterone excess mainly caused by aldosterone producing adenoma (APA). The prevalence of postoperative hyperkalemia was ranged from 10.5 % to 29.1 % in recent reports. However, there is no information on the prevalence of hyperkalemia in patients with primary aldosteronism (PA) underwent medical treatment. In this study, the prevalence and the risk factors of hyperkalemia after treatment including adrenalectomy and medication from data of a multi-center collaborative study in Japan (WAVES-J study) were investigated. Data from 424 patients with PA (185 male, 239 female, mean age 55 ± 11 yrs.) who underwent adrenal vein sampling (AVS) and were followed up after treatment in 9 centers were analyzed retrospectively. Of 424 patients, 142 patients were performed unilateral adrenalectomy (group A) and 282 patients were treated by medication (group B). In group B, 91% of patients took mineralocorticoid receptor antagonist after diagnosis. The prevalence of hyperkalemia (serum potassium >5.0 mEq/L) after treatment was significantly higher in group A (14/142, 9.9 %) than that in group B (9/282, 3.2%) (p<0.01). In group A, hyperkalemic patients were older (62 ± 6 vs. 52 ± 11 yrs.), had longer duration of hypertension (18 ± 10 vs. 8 ± 8 yrs.), higher serum creatinine (1.15 ± 0.55 vs. 0.71 ± 0.19 mg/dl), lower glomerular filtration rate (GFR) (52.0 ± 18.5 vs. 82.4 ± 18.4 ml/min/1.73m2) at the time of diagnosis compared with normokalemic patients (p<0.05). In group B, hyperkalemic patients were older (66 ± 5 vs. 55 ± 11), had higher percentage of male (78 vs. 39 %), higher plasma renin activity (0.84 ± 0.73 vs. 0.42 ± 0.34 ng/ml/h), higher serum creatinine (0.94 ± 0.29 vs. 0.72 ± 0.17), lower GFR (62.7 ± 15.4 vs. 77.9 ± 17.3) at the time of diagnosis compared with normokalemic patients (p<0.05). In conclusion, hyperkalemia was also experienced in patients underwent medical treatment though less common compared with that in patients underwent surgery. The risk factors of developing hyperkalemia after treatment were different between the patients treated by surgery and medication. Despite of the type of treatment, the occurrence of hyperkalemia should be considered in patients with PA after treatment.


Nothing to Disclose: NW, YS, HU, TI, YF, KK, TK, RS, AO, YM, TF, MT, TS, MN

25215 19.0000 FRI 590 A Hyperkalemia after Medical Treatment in Patients with Primary Aldosteronism: A Multi-Center Collaborative Study in Japan (WAVES-J study) 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 3:15:00 PM FRI 572-594 7662 1:15:00 PM Molecular Regulation and Diagnosis of Hyepraldosteronism (posters) Poster

Yuta Tezuka*1, Ryo Morimoto1, Yoshikiyo Ono1, Yoshitsugu Iwakura1, Yasuhiro Igarashi1, Masataka Kudo1, Masahiro Nezu1, Kei Omata2, Sadayoshi Ito1 and Fumitoshi Satoh1
1Tohoku University Hospital, Sendai, Japan, 2Tohoku University, Sendai, Japan



Magnesium (Mg) is an essential cofactor for many metabolic reactions, which homeostasis is maintained by bone, intestine and kidney. Primary aldosteronism (PA) can cause Mg deficiency because aldosterone inhibits reabsorption of urinary Mg in kidney. Moreover, hypomagnesemia may increase urinary potassium excretion and impair hypocalcemic-induced parathyroid hormone secretion.


This study aimed to evaluate serum Mg level (S-Mg) and urinary Mg excretion (U-Mg) in PA, and analyze the associations among Mg, plasma aldosterone concentration (PAC) and other variables.

Material & Method

We performed the retrospective study for 332 PA patients, who were diagnosed with PA by a couple of confirmatory tests, captopril challenge test and adrenal vein sampling at our center from April 2007 to September 2015, and measured S-Mg. Hypokalemia was actively corrected by prescribing potassium replacement.


The mean age of the participants was 53.2 ± 11.0 (mean ± SD) years and the percentage of male was 46.7%. The baseline parameters at the diagnosis were as follows: plasma renin activity 0.20 ±0.18 ng/ml/hr; PAC 27.7 ± 24.4 ng/dl; urinary aldosterone excretion (U-Aldo) 16.9 ± 13.6 μg/day; aldosterone-to-renin ratio 210.0 ± 246.9; S-Mg 2.22 ± 0.18 mg/dl; U-Mg 0.09 ± 0.04 g/day; fractional excretion of Mg (FEMg) 3.8 ± 2.0%; serum potassium (S-K) 4.08 ± 0.55 mM; urinary potassium excretion (U-K) 59.0 ± 30.7 mmol/day; estimated glomerular filtration rate 77.4 ± 16.2 ml/min/173m2. Using Spearman’s correlation, S-Mg was positively correlated with S-K (rs = 0.204, p = 0.0002). Among the participants (n = 128), who were measured U-Mg, U-Mg was positively correlated with U-K (rs = 0.336, p = 0.0001), PAC (rs = 0.251, p = 0.0042) and U-Aldo (rs = 0.348, p = 0.0001). Furthermore, FEMg in unilateral PA (n = 67) was significantly higher than that in bilateral PA (p = 0.024) as well as PAC (p < 0.001). In multiple regression analysis, PAC was an independent prognostic factor for U-Mg (β = 0.021, p = 0.02), and U-Mg was also an independent prognostic factor for U-K (β = 131.7, p = 0.019) after adjustment for PAC.


Our study implied that high level of aldosterone may increase U-Mg as well as U-K.


Nothing to Disclose: YT, RM, YO, YI, YI, MK, MN, KO, SI, FS

26468 20.0000 FRI 591 A Evaluation of Magnesium Metabolism in Primary Aldosteronism 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 3:15:00 PM FRI 572-594 7662 1:15:00 PM Molecular Regulation and Diagnosis of Hyepraldosteronism (posters) Poster

Daisuke Watanabe*, Midori Sasaki Yatabe and Atsuhiro Ichihara
Tokyo Women's Medical University, Tokyo, Japan


Primary aldosteronism (PA) is a clinical syndrome characterized by hypokalemia, hypertension, and impaired glucose tolerance (1). Diabetes in PA is characterized by the impairment of both insulin secretion and sensitivity, but the degree of contribution of these two factors is varied. Therefore, we evaluated insulin sensitivity and secretion in patients with PA using Homeostasis model assessment (HOMA) and the insulinogenic index (IGI) indices calculated from the results of a 75-gram oral glucose tolerance test (OGTT). These parameters were compared to those from patients with essential hypertension (EH). We analyzed 32 patients with PA. OGTT was performed before adrenalectomy in all patients, and also after the surgery in a subset of patients. The control EH group consisted of 21 essential hypertensive patients with a normal glucose tolerance. There were no significant differences in age, BMI, blood pressure, or lipid parameters between PA and EH group. Fasting glucose and plasma insulin levels of PA patients were higher than those of EH patients. In addition, PA patients had significantly higher stimulated plasma glucose levels after 120 min and lower plasma insulin levels after 30 min compared to EH patients. Insulin sensitivity, evaluated by QUICKI (0.36±0.03 vs. 0.38±0.03) and HOMA-IR (1.79±0.13 vs. 1.26±0.11), was more impaired in patients with PA compared to patients with EH, and IGI in patients with PA was significantly less than that of patients with EH (0.62±0.09 vs. 0.93±0.12). On the other hand, pancreatic β−cell function HOMA-βF (71.8±5.7 vs. 73.8±4.2 %) was similar between PA and EH patients. Serum potassium correlated inversely with HOMA-IR and positively with QUICKI in PA patients (r = 0.51, P < 0.05; r = 0.38, P <0 .05 respectively). In 5 PA patients, OGTT was also performed after unilateral adrenalectomy, and in these patients, insulin sensitivity evaluated by HOMA-IR significantly improved after the operation. Insulin action in patients with PA was characterized by insulin resistance associated with hypokalemia in addition to the impairment of early-phase secretory response to glucose. Therapeutic intervention aimed at correcting both potassium and aldosterone levels might improve insulin action in patients with PA.


Nothing to Disclose: DW, MSY, AI

24227 21.0000 FRI 592 A Evaluation of Insulin Sensitivity and Secretion in Primary Aldosteronism 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 3:15:00 PM FRI 572-594 7662 1:15:00 PM Molecular Regulation and Diagnosis of Hyepraldosteronism (posters) Poster

Tobias Akerstrom*, Per Hellman and Peyman Bjorklund
Uppsala University, Sweden


Unilateral primary aldosteronism is a prevalent and curable cause of hypertension. Aldosterone producing adenomas (APAs) represent a common form of this disease. These tumours can present as solitary adenomas or display surrounding multinodular hyperplasia, raising the question which nodules cause the increased aldosterone secretion. The recent development of an antibody directed against aldosterone synthase encoded by the CYP11B2 gene have aided in the diagnosis of aldosterone production. In this study we wanted to investigate a new form of diagnosing functionality, by direct measurement of aldosterone in tumour tissue. We included forty-seven APAs, an additional three nodules from multinodular unilateral PA, one breast tumour and one cortisol producing adenoma in the analysis. The cohort had previously been screened for mutations in KCNJ5, ATP1A1, ATP2B3, CACNA1D and CTNNB1. Immunohistochemistry for aldosterone synthase was performed. Aldosterone was measured in lysed tumour tissue using a chemiluminescent immunoassay used in routine clinical practice.

All tumors with known mutations contained aldosterone at higher levels than the negative controls. The aldosterone content showed a small but not significant decline with length of time in the freezer (r2= 0.036, p=0.23). No difference in aldosterone content was observed between APAs with G151R (n=13) and L168R KCNJ5 mutations (n=11). ATP1A1/ATP2B3 mutated tumours contained a significantly higher amount of aldosterone compared to KCNJ5 mutated tumours, 4.3 vs 1.3 pmol/mg (p=0.0041). In the multinodular adrenals, a higher level of aldosterone was observed in mutated nodules. In adrenals with low CYP11B2 expression low aldosterone content was observed. Importantly, of the tumours classified as non-mutated, 6/13 (46%) did not contain aldosterone, suggesting a non-functional nodule. In conclusion, this method demonstrated the differences between nodules and different mutations in terms of aldosterone release, implying a heterogeneous secretion pattern and may represent an easy and inexpensive way of diagnosing aldosterone production.


Nothing to Disclose: TA, PH, PB

27565 22.0000 FRI 593 A Tumor Tissue Aldosterone Content Measurement for Identification of Aldosterone Producing Nodules 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 3:15:00 PM FRI 572-594 7662 1:15:00 PM Molecular Regulation and Diagnosis of Hyepraldosteronism (posters) Poster

Marianne A. Grytaas*1, Anette Heie1, Melissa D. Jørstad2, Jarle T. Rørvik1, Thomas Arnesen2, Siri S. Strømsøy3, Håvard Rørvik1, Bjørn G. Nedrebø4, Joern V. Sagen2, Gunnar Mellgren2, Hrafnkell Thordarson1, Eystein S. Husebye2 and Kristian Løvås2
1Haukeland University Hospital, Bergen, Norway, 2University of Bergen, Bergen, Norway, 3University of Bergen, Bergen, 4Haugesund Hospital, Haugesund, Norway



Endocrine Society Guidelines from 2008 recommend confirmatory testing when primary aldosteronism (PA) is suspected, with subsequent adrenal vein sampling (AVS), followed by adrenalectomy in unilateral, and medical treatment in bilateral disease. Our aim was to describe clinical characteristics, diagnostic procedures and long-term outcomes for PA patients in Western Norway.

Material and methods

Data from all suspected PA patients investigated at Haukeland University Hospital in the period 1998-2012 were retrospectively evaluated for inclusion. Patients with verified PA after saline infusion testing (SIT) or who otherwise were considered highly likely of having PA, (elevated aldosterone/renin-ratio, hypokalemia and/or radiological evidence of adenoma or hyperplasia), were included. Clinical, biochemical, radiology findings, and AVS-reports were retrieved. All patients still alive by August 2014 were invited to a clinical follow-up visit.


One hundred and eight patients were included. The median age at PA-diagnosis was 54 years (range 30-85); the median duration of hypertension was 10 years (range 0-30). Eighty-five percent (85%) were hypokalemic. Eighty-two (76%) had PA confirmed by SIT. AVS was performed in 95 (88%) patients, of whom 50 (53%) had unilateral, and 20 (21%) bilateral disease; the remaining did not have representative AVS-results. For patients with representative AVS, the proportion bilateral disease increased from 17% (5/29) before 2008, to 37% (15/41) from 2008 onwards.           

Altogether, 68/108 (63%) were adrenalectomized, of whom 48 (71%) had AVS-confirmed unilateral disease. The remaining were operated based on radiological findings alone. Forty patients were medically treated.

Follow-up visits were performed for 72/108 (67%); median 72 months (range 27-186) after treatment. Blood pressure reductions were similar in adrenalectomized (n= 52) and medically treated (n=20) patients (26/8 vs. 20/16 mm Hg, p=ns). Among the adrenalectomized (n=52), eleven (21%) were cured of HT, thirty-four (65%) had improved, and seven (13%) showed no improvement of HT. Nine of the cured patients had available pathology reports, all showing adenomas. Among the patients not cured, 63% had adenomas and 37% hyperplasia. Using logistic regression analysis, only female sex significantly predicted cure of hypertension(p= 0.029).


AVS was performed in a high proportion of our patients, of whom the majority had unilateral disease and hypokalemia, indicating that patients with bilateral disease and milder PA may still be underdiagnosed. The cure rate of HT after adrenalectomy was low and associated with female sex.


Nothing to Disclose: MAG, AH, MDJ, JTR, TA, SSS, HR, BGN, JVS, GM, HT, ESH, KL

24887 23.0000 FRI 594 A Clinical Outcome after Treatment of Primary Aldosteronism in Western Norway 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Cardiovascular Endocrinology Friday, April 1st 3:15:00 PM FRI 572-594 7662 1:15:00 PM Molecular Regulation and Diagnosis of Hyepraldosteronism (posters) Poster

Cornelie D. Andela*1, Sabine M. Staufenbiel2, Sjoerd D. Joustra1, Alberto M. Pereira1, Elisabeth F.C. van Rossum3 and Nienke R. Biermasz1
1Leiden University Medical Center, Leiden, Netherlands, 2Erasmus MC, Rotterdam, Netherlands, 3Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands


In patients with adrenal insufficiency (AI) a higher hydrocortisone intake has been associated with more impairment in Quality of Life (QoL). Irrespective of age, sex and severity of AI the dosage of hydrocortisone is titrated around 20 mg/D in all adult patients with AI based on physical and mental signs and symptoms. However, until now it is unknown whether these QoL impairments are related to increased systemic cortisol exposure. Measurement of hair cortisol levels (CORThair) can be used to assess chronic systemic cortisol exposure. This study aimed to explore whether QoL in patients with AI is associated with CORThair and daily hydrocortisone intake. We performed a cross-sectional study in 120 patients with AI on stable hydrocortisone replacement, in whom hair samples and QoL data were collected. CORThair were measured with ELISA, and QoL was assessed with validated questionnaires (SF-36, EQ-5D, HADS, MFI-20). Patients reported impairments in 14 of 15 QoL subscales (p < .001). More impairment in physical aspects of QoL correlated with higher CORThair and higher daily hydrocortisone intake (p < .05), an effect that was more pronounced in female patients. Regression analyses with CORThair and hydrocortisone intake in the same model revealed a significant negative contribution of higher hydrocortisone intake on physical aspects of QoL (p ≤ .046), whereas no significant independent contribution was found for CORThair. The present study showed that patients with AI report several impairments in QoL which are associated with hydrocortisone intake, and to a lesser extent reflected by chronic systemic cortisol exposure as measured by hair cortisol. This suggests that QoL impairments in patients with AI are not per se the effect of overtreatment with hydrocortisone.


Nothing to Disclose: CDA, SMS, SDJ, AMP, EFCV, NRB

25520 1.0000 FRI 394 A Quality of Life in Patients with Adrenal Insufficiency Correlates Stronger with Hydrocortisone Dosage, Than with Long-Term Systemic Cortisol Levels 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Adrenal HPA Axis Friday, April 1st 3:15:00 PM FRI 394-408 7668 1:15:00 PM Glucocorticoid Actions (posters) Poster

Sheila Piccoli Garcia*, Camila Viecceli, Thiza Massaia Londero, Ana Marina da Silva Moreira, Fabiola Costenaro, Iuri Martin Goemann, Gustavo Cipriani, Ticiana Costa Rodrigues and Mauro Antonio Czepielewski
Hospital de Clínicas de Porto Alegre, UFRGS, Porto Alegre, Brazil


Glucocorticoid (GC) excess has multiple effects on bone health and it is a known cause of secondary osteoporosis. Pathogenesis is multifactorial and not restricted to loss of bone mineral density (BMD). Sustained hypercortisolism leads to an excessive resorption and an impaired bone formation. GC also reduces vitamin D action, inhibits renal tubular calcium reabsorption and impairs secretion of several hormones (gonadotropins and growth hormone). The prevalence of fractures in Cushing’s Syndrome (CS) is estimated in 70% and the role of vitamin D deficiency in these patients remains unclear. To investigate factors associated with BMD loss leading to fracture we analyzed 34 patients diagnosed with CS between 2006 and 2015. Patients underwent clinical, radiological and laboratorial evaluation at diagnosis. BMD was evaluated by dual energy X-ray absorptiometry (DXA). Fractures were confirmed on radiographs. T-test and X² test were used to compare parameters between groups. Logistic regression analysis was performed to assess variables associated with bone fracture. Out of 34 patients (mean age 34 ±14y, 76% female, 91% Caucasian) with CS diagnosis (33 pituitary and 1 adrenal adenoma), 11 (32.4%) presented with fracture at diagnosis, particularly at vertebral body. Patients with bone fracture were older (46 vs 30 years, p=0.004) and had higher hypogonadism prevalence (54 vs 12%, p= 0.033) when compared with patients without fractures. No difference were found in bone mass according to gender, ethnic group, hypercortisolism levels (urinary free cortisol, morning or midnight cortisol) or vitamin D levels. Hypogonadism (OR 8.40 CI 95% 1.26 to 56.07 p=0.028 was associated with bone fracture. In subjects with CS, hypogonadism seems to play a determinant role in bone fracture, independent of severity of hypercortisolism or others confusion factors.


Nothing to Disclose: SP, CV, TML, AMDSM, FC, IMG, GC, TCR, MAC

27184 2.0000 FRI 395 A Cushing's Syndrome and Bone: An Analysis of Related Factors Leading to Increased Risk of Fractures 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Adrenal HPA Axis Friday, April 1st 3:15:00 PM FRI 394-408 7668 1:15:00 PM Glucocorticoid Actions (posters) Poster

Andrea Rebellato1, Eleonora Avenatti2, Marialberta Battocchio1, Andrea Iannaccone2, Francesca Dassie1, Franco Veglio2, Alberto Milan2 and Francesco Fallo*1
1Padova University Hospital, Padova, Italy, 2University of Torino, Torino, Italy


Cushing’s syndrome (CS) is characterized by a cluster of systemic manifestations, including abdominal adiposity, arterial hypertension, impaired glucose tolerance/diabetes, dyslipidaemia and pro-thrombotic state. All these features are associated with increased cardiovascular risk. The impact of the hemodynamic overload on left ventricular mass (LVM) in patients with CS is not well known. The aim of our study was  to assess the presence of cardiac organ damage in relation to the global pressure burden, assessed with 24hour blood pressure (BP) monitoring, in patients with newly diagnosed CS. Twenty-five patients (4 M, 21 F) with CS (21 pituitary-dependent and 4 with cortisol-producing adenoma) underwent echocardiography in order to assess cardiac morphology (left ventricular hypertrophy) and geometry (relative wall thickness-RWT). BP levels were tested in office and using 24h ambulatory blood pressure monitoring (ABPM). Twenty-five subjects similar for demographic characteristics and 24h BP, were used as controls.  Patients with CS were similar to controls by age (44±11 vs 50 ±13 years, p 0.10), sex (females 84%, p 1.0), mean 24h BP (102±10.9 vs 104.3 ± 14.2 mmHg, p 0.7), and body size (body weight 80 (95%CI:65-86) vs. 70 (62-79) kg, p 0.13; body mass index 24.6 (22.6-29) vs. 27.1 (25.4-32.4) kg/m2, p=0.06). There was an increased LVM indexed by height2.7 (44.4 ±14.7  vs 36.9 ±10 g/m2.7, p=0.03) and an increased RWT (0.46 ±0.07 vs 0.41 ± 0.08, p=0.02) in CS patients compared to controls, leading to a higher prevalence concentric hypertrophy (36% vs 4%, p<0.05) and concentric remodelling (36% vs 28%), respectively. The prevalence of non-dipping BP profile at ABPM in CS patients was greater than that of controls (56% vs 16%, p <0.05), with no significant association with LVM or geometry. 24h urinary cortisol was not associated with LVM (r= 0.1, P 0.5) or RWT (r= 0.02 P 0.89) in the CS group. Conclusions: LVM and the concentric pattern of left ventricle are relatively independent from 24h BP load and profile (dipping / non-dipping) in CS patients.


Nothing to Disclose: AR, EA, MB, AI, FD, FV, AM, FF

24053 3.0000 FRI 396 A Left Ventricular Geometry and 24h Blood Pressure Profile in Cushing's Syndrome 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Adrenal HPA Axis Friday, April 1st 3:15:00 PM FRI 394-408 7668 1:15:00 PM Glucocorticoid Actions (posters) Poster

Alexandria Ajoa Atuahene*, Tin Htwe Thin, Vanessa Cohen, Nyima Yangdhar, Irene Lee, Michael J. Donovan and Eliza B. Geer
Icahn School of Medicine at Mount Sinai, New York, NY


Background:  Obesity is characterized by chronic low-grade inflammation, which in turn is associated with comorbidities including diabetes and cardiovascular disease. A cardinal feature of obesity-related inflammation is recruitment of macrophages within adipose tissue (AT), typically in a formation known as crown-like structures (CLS). Glucocorticoids (GCs) have well-known anti-inflammatory actions. Paradoxically, chronic excess GC exposure results in the development of conditions that are considered pro-inflammatory, including obesity and insulin resistance. Some data suggest that patients with Cushing’s disease (CD), a state of chronic GC excess, have elevated circulating pro-inflammatory cytokines compared to BMI-matched controls. The mechanisms behind enhanced systemic inflammation in patients with CD remain unknown.

Objective: To investigate the effect of chronic GC exposure on AT inflammation by quantifying macrophage infiltration in AT biopsies from patients with active CD compared to BMI-matched controls.

Methods:  Immunohistochemistry on subcutaneous abdominal AT biopsies from 6 active CD patients were compared to 9 overweight BMI-matched controlsTo qualitatively assess for varying architectural phenotypes within the AT, antibodies for vimentin, an intermediary filament that stains mesenchymal cells, and CD31, an endothelial marker, were used. Macrophage infiltration was evaluated by the presence of pan-macrophage CD68 and activated macrophage CD163-positive cells. 

Results:  The 6 enrolled CD patients (6 female) had a median age of 30.5 years (range 19-53), median BMI of 34.5 kg/m2 (range 29.6-61.8), and median waist circumference (WC) of 103.4 cm (range 98.4-157.5).  The 9 control patients (5 female) had a median age of 27 years (range 24-41), median BMI of 31 kg/m2 (range 21.8-37.5), and median WC of 89.1 cm (range 77.5-105.4).  There were no differences in mean age (p=0.72) or BMI (p=0.06) between CD patients and controls, but CD patients had a higher mean WC vs. controls (p=0.02). Analysis of AT immunohistochemistry revealed a qualitative increase in the expression of Vimentin and CD31 in AT from CD patients vs. matched controls.  100% of AT samples evaluated from CD patients (N=5) exhibited a focal to diffuse increase in CD68+ macrophages, with CLS (i.e. aggregates of CD68+ macrophages surrounding adipocytes), compared to 1 out of 9 (11%) control AT samples with focal CD68+ cells and no CLS. 

Conclusions:  Our preliminary qualitative data demonstrate that, compared to BMI-matched controls, AT from patients with CD are characterized by an influx of CD68+ macrophages and a possible increase in vimentin and vascular networks. These findings suggest that chronic excess GC exposure has a paradoxical pro-inflammatory morphologic impact on AT. Additional studies are ongoing to quantify and further phenotype AT macrophage content in patients with CD.


Disclosure: EBG: Study Investigator, Chiasma, Study Investigator, Novartis Pharmaceuticals, Consultant, Cortendo, Consultant, Chiasma, Consultant, Ipsen, Consultant, Pfizer, Inc., Study Investigator, Cortendo. Nothing to Disclose: AAA, THT, VC, NY, IL, MJD

24947 4.0000 FRI 397 A Glucocorticoid Regulation of Adipose Tissue Macrophages:  Evidence from Cushing's Disease 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Adrenal HPA Axis Friday, April 1st 3:15:00 PM FRI 394-408 7668 1:15:00 PM Glucocorticoid Actions (posters) Poster

Sven Michael Danneberg*1, Birgit Harbeck1, Christian S Haas1, Amir Hossein Rahvar1, Hendrik Lehnert2, Peter Kropp3 and Heiner Moenig4
1University of Lübeck, Lübeck, Germany, 2University of Luebeck, Luebeck, Germany, 3University of Rostock, Rostock, Germany, 4Christian-Albrechts- University Kiel, Kiel, Germany


Hydrocortisone (HC) substitution is essential in the treatment of patients with adrenal insufficiency (AI). Current replacement regimens however incompletely mimic the physiological circadian rhythm of cortisol secretion, thereby resulting in subclinical temporary hypo-and hypercortisolism. Several studies point towards impairment of cognitive functions under these conditions, in part due to affected catecholamine secretion.

To evaluate the influence of long-term vs. short-term cortisol replacement therapy on the adrenomedullary system and cognitive functions.

Fourteen patients with primary or secondary AI were divided into two groups, depending on the duration of disease and HC replacement therapy (less or at least 15 years). All subjects underwent standardized neurocognitive testing; in addition, cortisol and catecholamine levels as well as physiological parameters and quality of life (QoL) were assessed.

Patients with HC replacement therapy >15 years (n=7) received significantly higher equivalent glucocorticoid (GC) doses than those with a shorter lasting therapy (n=7; p=0.048). Neuropsychological tests, QoL, physiological parameters and cortisol levels did not differ significantly between both groups. While epinephrine concentrations in plasma were subnormal in both groups, norepinephrine levels were in the normal range but significantly lower in patients on short-term HC replacement therapy (p= 0.025). 

We showed that duration of cortisol replacement therapy may have an impact on catecholamine release, but does not seem to have an impact on cognitive functions and QoL.


Nothing to Disclose: SMD, BH, CSH, AHR, HL, PK, HM

27400 5.0000 FRI 398 A Cognitive Function and Catecholamine Secretion in Short- and Long-Term Hydrocortisone Replacement 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Adrenal HPA Axis Friday, April 1st 3:15:00 PM FRI 394-408 7668 1:15:00 PM Glucocorticoid Actions (posters) Poster

Esra Suheda Hatipoglu*1, Arzu Yavuz2, Mutlu Niyazoglu3, Nilufer Alcalar4 and Yalcin Hacioglu3
1Liv Hospital, Istanbul, Turkey, 2Ministry of Health’s Kocaeli Derince Education and Research Hospital, Kocaeli, Turkey, 3Istanbul Teaching and Research Hospital, Istanbul, Turkey, 4Istanbul Medical School, Istanbul University, Istanbul, Turkey


Abstract:Stress responses involve complex mechanisms including various behavioural and endocrinologic adaptations. Pregnancy itself may be also a state of stress for the mother. Moreover, hypothalamus-pituitary-adrenal axis (HPA) is subject to certain physiological changes during pregnancy. Although gestation is a state of hypercortisolism, response to HPA is hampered during late pregnancy. How HPA changes during pregnancy affect psychologic status of females is still controversial (1,2). Moreover previous studies have evaluated earlier periods of gestation rather than the late pregnancy.

To study the effects of physiologic alterations in cortisol mileu on mood changes during late pregnancy, we prospectively evaluated 75 healthy pregnant subjects after 36 weeks of gestation and at 3-4 weeks of postpartum period. During each visit blood samples were taken to determine the levels of total cortisol (TC) and cortisol binding globulin (CBG) at 8 a.m. Free cortisol (FC) was calculated using Coolen's equation (3) and free cortisol index (FCI) was defined as serum TC/CBG. Concurrently, status of depression, anxiety and stress were graded using Beck Depression Inventory (BDI),  Beck Anxiety Inventory (BAI) and Percieved Stress Scale (PSS). Additional demographic data and support mechanisms of each subject were also questionnaired.

The mean age of the participants was 28.5±4.9 years and the median week of gestation was 39 [IQR: 38-40]. After gestation TC levels decreased from 130.4 [IQR: 101.1-167.5] to 53 [IQR: 41.7-73.1] nmol/l, FC levels from 22 [IQR: 14.2-38.3] to 9.9 [IQR: 6.4-15.2] nmol/l and FCI from 28.9 [IQR: 21.6-40.1] to 25.5 [IQR: 16.9-36.8] nmol/mg (p<0.001 for all). At late pregnancy and early postpartum period the median score on BDI was 9 [IQR: 6-14.5] and 7 [IQR: 3-11.5] (p=0.006), on BAI was 15 [IQR: 9-19] and 6 [IQR: 2-12] (p<0.001) and on PSS was 23 [IQR: 19-28.5] and 22 [IQR: 17-26] (p=0.02), respectively. Both at late pregnancy and postpartum period neither TC levels nor FC measures were correlated with the concurrent scores on BDI, BAI and PSS. However higher FC levels during late pregnancy were associated with lower scores on stress and depression at early postpartum period, albeit the latter was not statistically significant (r= -0.2, p=0.03 and r= -0.2, p=0.06). Additionally, as FCI increased during late pregnancy both the scores on stress and depression decreased during early postpartum period (r= -0.4, p=0.02 and r= -0.3, p=0.01).

We speculate that physiologic hypercortisolism during late pregnancy may not add a burden on psychology of pregnant cases. Moreover increased cortisol levels during  latest terms of pregnancy may cause indirect and ultimate alterations, having protective effects on mood of female cases during postpartum period.


Nothing to Disclose: ESH, AY, MN, NA, YH

24634 6.0000 FRI 399 A The Impact of Physiological Changes in Hypothalamus-Pituitary-Adrenal Axis on Psychological Status of Females during Late Pregnancy and Pospartum Period 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Adrenal HPA Axis Friday, April 1st 3:15:00 PM FRI 394-408 7668 1:15:00 PM Glucocorticoid Actions (posters) Poster

Brent S. Abel*1, Susmeeta T. Sharma2, Raven N. McGlotten3, Pooja Raghavan1, Monica C. Skarulis1 and Lynnette K. Nieman2


Background: The mechanism underlying the weight gain experienced by most patients with Cushing’s syndrome (CS) is not fully understood. A past study of 18 CS patients found that resting energy expenditure (REE) adjusted for lean body mass (LBM) did not differ from healthy controls, and a follow-up study in 10 of these patients after successful surgery found their REE unchanged from before (1-2). We aimed in this study to test the reproducibility of these findings and examine whether hormonal changes during the recovery process may influence changes to REE.

Methods: Patients admitted to the NIH Clinical Center for the evaluation and treatment of active CS are eligible for study. During an inpatient admission, baseline REE is measured by indirect calorimetry (TrueOne 2400) after waking, at rest, and prior to receiving any medications. Baseline body composition is measured by DXA and blood is drawn in the morning after a 12 hour fast. These measurements are repeated at inpatient admissions at 6 and 12 months after successful surgery. Data were compared between two intervals by paired t-test or Wilcoxon signed rank test and are presented as mean ± SEM.

Results: 29 patients have enrolled in this study, of which 11 have completed a 6 month follow-up visit after surgical treatment. These 11 patients are 82% female (n=9) and 37± 4 years of age. All 11 subjects had ACTH dependent CS (11 pituitary, 1 bronchial carcinoid). REE adjusted for LBM significantly increased 32 ± 6% 6 months after surgery (pre: 27 ± 2 kCal/d/kg, post: 34 ± 1 kCal/d/kg, p<0.001). Unadjusted REE also significantly increased (pre: 1284 ± 98 kCal/d, post: 1644 ± 89, p<0.001). LBM did not significantly change (pre: 49 ± 4 kg, post: 49 ± 3, p=0.905), but BMI (pre: 37 ± 4 kg/m2, post: 34 ± 3, p=0.012) and percent body fat (pre: 46 ± 3%, post: 42 ± 3, p=0.025) decreased. Patients lost an average of 9 ± 3 kg, or 7.1 ± 2.5%, of body weight (p=0.022). Excluding one patient on levothyroxine, TSH (pre: 1.2 ± 0.2, post: 2.9 ± 0.4, p<0.001), total T3 (pre: 78 ± 5, post: 130 ± 5, p<0.001), and free T4 (pre: 0.9 ± 0.1, post: 1.1 ± 0.0, p=0.038) increased. We await additional data on the 12 month visits.

Discussion: Weight loss observed in the 6 months after successful surgery in CS patients is associated with higher REE and higher circulating levels of TSH and thyroid hormones. This suggests that decreases in REE associated with lower thyroid hormone levels may contribute to weight gain in CS prior to surgery. The effect of cortisol excess on binding proteins and deiodinase activity is being examined. It is unclear if this effect will persist at one year after cure of CS or whether the expected decrease in REE with further weight loss will be observed.


Disclosure: LKN: Investigator, HRA Pharma. Nothing to Disclose: BSA, STS, RNM, PR, MCS

25962 7.0000 FRI 400 A Increased Resting Energy Expenditure Following Surgical Cure in Patients with Cushing's Syndrome 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Adrenal HPA Axis Friday, April 1st 3:15:00 PM FRI 394-408 7668 1:15:00 PM Glucocorticoid Actions (posters) Poster

Raven N. McGlotten*1, Susmeeta T. Sharma2 and Lynnette K. Nieman2


Background: Diabetes mellitus (DM) is seen in ~40-45% of patients (pts) with Cushing’s Syndrome (CS)(1). The reversibility of abnormal glucose tolerance after successful surgical cure of CS is not well understood. We evaluated insulin sensitivity shortly after surgical remission of CS and on long-term follow-up (f/u).

Methods: We retrospectively studied pts with CS admitted to our protocols at the NIH Clinical Center from 1/2012 to 10/2015. Ectopic ACTH syndrome (EAS) or Cushing’s disease (CD) was diagnosed based on pathology. Pts were considered to have DM based on a HbA1c ≥6.5%, a fasting plasma glucose (FPG) ≥125 mg/dl, or if they were on oral hypoglycemic medications (meds) or insulin therapy. After surgery, pts were inpatients for 7-15 days; hydrocortisone (HC) (10-12mg/m2) was started after remission was confirmed. Usually insulin was used for glycemic control until discharge. Pts returned every 3-6 months (mo) for f/u and HC dose was adjusted based on weight and recovery of adrenal function. Resolution of DM was defined as HbA1c<6.5% off all meds.

Results: 50 pts achieved remission after tumor resection. At baseline (BL), 15 (10 female) had a diagnosis of DM (13=CD, 2=EAS); 10 pts took oral antidiabetic meds and 5 took insulin 0.3-1.3units/kg/d. BL HbA1c was 6.3-9.4%. Evaluable pre and post-operative insulin usage data was available for 1st week after surgery in 11 pts: 2 pts previously on oral meds did not need insulin, 9 had a decrease in daily insulin dose, with 5 requiring none by day 7.

Of the 15 pts, at discharge, 4 required no DM meds, 4 had a decrease in insulin or med dosage, and 5 had no change to DM regimen. Despite overall decreased insulin use after surgery, 2 pts were discharged on higher med doses than BL due to uncontrolled DM on admission (HbA1c: 9.4, 7.6%).

8/15 pts were seen for 3-mo f/u. Of these, 4 were on no DM meds, insulin dose decreased in 2, while 2 pts had no change in regimen. HbA1c decreased in 7 and was unchanged in one. FPG decreased in all. By 6 mo, 6/8 took no meds, one was on a decreased regimen from BL, and one had no change. HbA1c was <6.5% (5-6.2%) in all. 7/8 came back for 12 mo f/u. Of these, 6 were on no meds or on decreased regimen with HbA1c and FPG in non-diabetic range. HbA1c (6.6%) increased in one pt due to over-replacement with HC but normalized by 18 mo.

2/3 pts first seen at 6 mo f/u were on no DM meds with HbA1c and FPG in non-diabetic range, the third pt had a decrease in DM regimen with decreasing HbA1c (6.8%) and FPG levels. One pt only seen at 9 mo had a normal HbA1c and FPG off all meds. 3 pts have not yet had a f/u after recent surgery. Overall, DM resolved in 10/12 pts with f/u by 3-18 mo. 

Conclusion: Insulin sensitivity and diabetic regimen can change rapidly, as early as the 1st week, after surgical remission of CS. We recommend close monitoring of blood glucose levels (especially in the 1st month), pt education and long-term f/u of these pts for optimal management of DM post-remission of CS.


Disclosure: LKN: Investigator, HRA Pharma. Nothing to Disclose: RNM, STS

26703 8.0000 FRI 401 A Change in Glucose Tolerance with Surgical Remission of Cushing's Syndrome 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Adrenal HPA Axis Friday, April 1st 3:15:00 PM FRI 394-408 7668 1:15:00 PM Glucocorticoid Actions (posters) Poster

Mesut Savas*1, Vincent L. Wester1, Ramon H.M. Dykgraaf2, Erica L.T. van den Akker1, Jolien W. Roos-Hesselink2, Arianne B. Dessens2, Ronald P. Stolk3, Yolanda B. de Rijke2 and Elisabeth F.C. van Rossum1
1Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands, 2Erasmus MC, Rotterdam, Netherlands, 3University Medical Center Groningen, Groningen, Netherlands


Introduction - Turner syndrome (TS), the most prevalent genetic disorder in females, originates from a (partial) loss of one X-chromosome and causes a wide range of comorbidities. In addition to psychological stress, many patients with TS suffer from cardiometabolic derangements and cardiovascular diseases, which has remarkable resemblances to a pathological hypercortisolistic state. Therefore we investigated whether TS patients are exposed to increased long-term cortisol concentrations as measured in scalp hair, and whether hair cortisol concentrations (HCC) are associated with metabolic parameters and experienced psychological stress in TS patients.

Methods - We collected scalp hair samples in 66 TS patients, and 216 sex-matched adult controls from the large population-based LifeLines cohort study. Subjects who used systemic and/or topical on scalp corticosteroids were excluded from the analysis. The proximal 3 cm of the hair samples, corresponding to cortisol exposure in the preceding three months, were processed before cortisol was extracted in methanol and further purified using solid phase extraction. HCC were subsequently quantified using LC-MS/MS. Anthropometry, fasting biochemical metabolic parameters were collected, and psychological stress was assessed using the Perceived Stress Scale (PSS) questionnaire.

Results - HCC could be quantified in 55 TS patients and in 186 adult female controls (median age: 31.0 and 41.0 years, respectively). TS patients had significantly higher long-term cortisol levels than controls (mean [95% CI]: 3.38 [2.61-4.37] vs. 2.33 [2.17-2.51], P<0.001, adjusted for age). In a stratified analysis, increased HCC appeared to be most pronounced in TS patients with isochromosome-Xq variant, but there was no significant difference between different karyotypes. In TS patients, HCC were positively associated with total cholesterol levels (standardized β=0.318, P=0.049, adjusted for age and BMI), but not with blood pressure, BMI, fasting triglycerides, LDL- and HDL-cholesterol, or fasting glucose. PSS-scores in TS patients were relatively high (mean (SD): 24.8 (±8.6)) and tended to increase with HCC, albeit not statistically significant.

Conclusions - Compared to sex-matched population controls, Turner syndrome patients are chronically exposed to higher systemic cortisol levels, which is associated with increased total cholesterol levels, and potentially contributes to an elevated cardiovascular disease risk.


Nothing to Disclose: MS, VLW, RHMD, ELTV, JWR, ABD, RPS, YBD, EFCV

27341 9.0000 FRI 402 A Increased Long-Term Cortisol Exposure in Turner Syndrome and Its Associations with Metabolic and Psychological Parameters 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Adrenal HPA Axis Friday, April 1st 3:15:00 PM FRI 394-408 7668 1:15:00 PM Glucocorticoid Actions (posters) Poster

Amit Tirosh*1, Maya Beth Lodish1, Charalampos Lyssikatos2, Elena Belyavskaya3, Richard A. Feelders4 and Constantine A Stratakis1
1National Institutes of Health, Bethesda, MD, 2Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), National Institutes of Health (NIH), Bethesda, MD, 3Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), National Institutes of Health (NIH)., Bethesda, MD, 4Erasmus Medical Center, Rotterdam, Netherlands


Cushing's syndrome has been associated with hypercoagulability, attributed to an increase in endogenous procoagulants (e.g., von-Willebrand factor [vWF], Factor VIII [FVIII] and fibrinogen), and antifibrinolytics (plasminogen activator inhibitor-1 [PAI-1] and α2-antiplasmin), despite a concomitant increase in endogenous anticoagulants (protein C [PC] and protein S [PS], and antithrombin III [AT-III]). However, a large multicenter study (1) reported that post-operative venous thrombo-embolic events occurred only among patients with Cushing's disease (CD), and not among adrenal CS (AdCS) patients.

To better understand this pathophysiological mechanism, we evaluated several key coagulation elements in patients with CS before and after intervention and in a control group.

This was a prospective observational study, including 85 consecutive patients, aged ≥18 years, that were admitted to the National Institutes of Health for evaluation of suspected CS. Coagulation profile was taken before and 6-12 months following surgery (either transsphenoidal surgery or adrenalectomy). The patients were divided into three groups: CD group (n=22) including patients with pathologically proven CD; AdCS group (n=21) including patients that underwent adrenalectomy due to confirmed autonomous cortisol secretion from the adrenal(s); and a control group (n=42) including patients that were screened for CS, and were found to be normocortisolemic.

Before intervention, patients with CD had higher mean UFC levels and PM plasma cortisol levels compared with the AdCS group, and both had higher levels compared with the control group (p<0.001 each). At baseline, patients with CS had higher levels of FVIII (p=0.04) and vWF (p=0.02) compared with controls, as well as higher PC (p=0.03) and AT-III (p=0.02) levels. Both PC and AT-III decreased significantly after intervention (p values of 0.02 and 0.006, respectively). Patient's with CD had higher levels of vWF (p=0.03) and AT-III (p=0.01) before surgery than patients with AdCS. Moreover, patients with CD had decrease in AT-III (p=0.016) and PC levels (p=0.032) after intervention, whereas patients with AdCS did not show similar dynamics. UFC levels correlated with AT-III (r=0.5, p<0.001), PC (r=0.4, p=0.008), PS (r=0.5, p<0.001), vWF (r=0.3, p=0.01), FVIII (r=0.5, p=0.001), and aPTT (r= -0.5, p=0.001). Decrease in UFC levels following intervention correlated positively with the dynamics of FVIII (r=0.8, p<0.001), and PC (r=0.6, p=0.02).

In conclusion, our data shows that patients with CD experienced significant decreases in PC and AT-III after intervention, a finding that was not present among patients with AdCS, and might be explained by higher cortisol levels in CD patients. This study also confirmed prior findings that patients with CS have an increase in endogenous procoagulants and anticoagulants, as well as in antifibrinolytics.


Nothing to Disclose: AT, MBL, CL, EB, RAF, CAS

27006 10.0000 FRI 403 A Coagulation Profile Dynamics in Adult Patients with Cushing's Syndrome (CS) Differ Depending on the CS Etiology and Cortisol Levels - a Prospective Observational Comparative Study 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Adrenal HPA Axis Friday, April 1st 3:15:00 PM FRI 394-408 7668 1:15:00 PM Glucocorticoid Actions (posters) Poster

Aziz Topaloglu1, Serkan Yener2, Murat Uyar1 and Abdurrahman Comlekci*2
1Sifa University Medical School, Izmir, Turkey, 2Dokuz Eylul University Medical School, Izmir, Turkey


Objective & Aim: Exogenous corticosteroid administration is a risk factor for intra ocular pressure (IOP) elevation, glaucoma and cataract. However, ophthalmologic complications are less common in patients with endogenous hypercortisolism (EHC). No study has yet measured the diurnal IOP variation, visual field parameters and lens opacity scores in patients with EHC and compared with the age matched controls.

Methods: 30 patients and 27 subjects without clinical or laboratory evidence of EHC were enrolled. We identified Cushing’s disease in 12, overt adrenal Cushing’s syndrome in 9 and subclinical Cushing’s syndrome in 9 subjects. All subjects underwent a full ophthalmological examination including best corrected visual acuity, slit-lamp assessment of lens opacities using the LOCSIII, slit-lamp gonioscopy, intraocular pressure measurement, fundus examination and diurnal IOP evalaution at 8:00 a.m., 12:00 p.m. and 4:00 p.m.

Results: Briefly, results of this study revealed no significant differences between the groups. We also did not observe a clear relationship between the prevalence of ophthalmological disturbances and the severity of the EHC (subclinical Cushing’s vs. overt Cushing’s syndrome).

Discussion: We attribute these results to relatively low potency of hypercortisolism in endogenous Cushing syndrome when compared to exogenous Cushing syndrome. Depending on the duration of exposure, total daily dose and the glucocorticoid type, glucocorticoid drugs are usually associated with more rapid development of cushingoid features. Additionally, there might be a referral bias for our patients when compared to the general population. Patients in this study have been evaluated in a tertiary health care unit by endocrinologists. This might be associated with early diagnosis, definitive treatment and the reduction of hypercortisolemia related co-morbidities.


Nothing to Disclose: AT, SY, MU, AC

26079 11.0000 FRI 404 A Diurnal Intraocular Pressure Measurements, Glaucoma and Cataract Evaluations in Patients with Endogenous Hypercortisolism 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Adrenal HPA Axis Friday, April 1st 3:15:00 PM FRI 394-408 7668 1:15:00 PM Glucocorticoid Actions (posters) Poster

Vincent L. Wester*1, Gerard Noppe2, Mesut Savas1, Ronald P. Stolk3, Erica L.T. van den Akker1, Yolanda B. de Rijke2 and Elisabeth F.C. van Rossum1
1Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands, 2Erasmus MC, Rotterdam, Netherlands, 3University Medical Center Groningen, Groningen, Netherlands


Introduction: Over the past decade, measurements of hair cortisone (HairE) and cortisol (HairF) have emerged as a method to estimate long-term glucocorticoid exposure over periods up to months of time. Increased hair glucocorticoids have been associated with unfavorable cardiometabolic health, physical and psychological stressors, and psychopathology. Little is known about the influence of local corticosteroid treatment and major life events on long-term glucocorticoid levels.

Subjects and methods: We collected scalp hair samples from the posterior vertex in 295 adult participants of LifeLines (median age: 42 yrs, 75% females), a large prospective population based cohort study in the north of the Netherlands (www.LifeLines.nl). Approximately 20 mg of the proximal 3 cm of hair was weighed, and washed using isopropanol. We extracted steroids in methanol, followed by solid phase extraction. HairE and HairF were quantified on a Waters Xevo TQ-S LC-MS/MS system. In all subjects, we performed anthropometry, and collected the long-term difficulties inventory (LDI), list of threatening experiences (LTE) and a questionnaire about hair characteristics and corticosteroid use. Fasting metabolic values were available in 97% of participants.

Results: HairE and HairF could be quantified in 98 and 90% of samples, respectively. In multiple linear regression, both HairE and HairF increased with age, male sex, black hair color, and occurrences of perspiration on the scalp, and decreased with hair washing frequency (P<0.05). Only HairE was increased with brown hair color and fasting glucose (P<0.05). HairE was lower in users of systemic corticosteroids (4.1 vs. 8.5 pg/mg hair, P<0.001), and in individuals who only used local corticosteroids (7.1 vs. 8.5, P=0.028). Life events (LTE) were associated with higher HairF (simple standardized β: 0.131, P=0.039), but this association may be mediated by the frequency of perspiration on the scalp (adjusted β: 0.068, P=0.306). In this subpopulation of the LifeLines cohort, hair glucocorticoids were not independently associated with BMI, waist circumference, blood pressure or fasting lipids.

Conclusion: HairE can be a useful marker to detect mild adrenal suppression due to corticosteroid use in the general population, even when only inhaled, nasal or topical corticosteroids are used. These findings suggest that commonly used local corticosteroids induce systemic effects.


Nothing to Disclose: VLW, GN, MS, RPS, ELTV, YBD, EFCV

24282 12.0000 FRI 405 A Long-Term Glucocorticoids Measured in Hair Are Influenced by Local Corticosteroid Treatment: the LifeLines Cohort Study 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Adrenal HPA Axis Friday, April 1st 3:15:00 PM FRI 394-408 7668 1:15:00 PM Glucocorticoid Actions (posters) Poster

Camila Viecceli*1, Thiza Massaia Londero1, Sheila Picolli Garcia2, Ana Marina da Silva Moreira1, Fabiola Costenaro1, Iuri Martin Goemann1, Gustavo Cipriani1, Ticiana Costa Rodrigues1 and Mauro Antonio Czepielewski3
1Hospital de Clínicas de Porto Alegre, UFRGS, Porto Alegre, Brazil, 2Hospital de Clínicas de Porto Alegre, Porto Alegre, 3Faculdade de Medicina UFRGS, Porto Alegre, Brazil


Diabetes mellitus (DM) plays an important role in morbidity and death of patients with uncontrolled hypercortisolism. Some authors define DM in CS as a specific type of diabetes secondary to endocrinopathy’, although others judge it as a classical form of type 2 DM. Glucocorticoid (GC) excess causes pancreatic beta cell dysfunction and insulin resistance, which correlates with hypercortisolism level. If Cushing’s disease (CD) remission implicates on DM resolution remains unclear. To assess DM prevalence in CD patients and DM resolution rate after one-year remission of CD we performed a retrospective cohort of 108 patients diagnosed with CD between 1987 and 2014. Patients underwent clinical and metabolic evaluation at diagnosis and 1-year after transsphenoidal surgery (TSS). CD remission criteria after TSS were: cortisol <3 mcg/dl on the 1mg-overnight test, normal urinary free cortisol (UFC) and/or adrenal insufficiency with GC dependence for 6 months. DM resolution criteria were HbA1c <6.5% and fasting glucose <126mg/dl without antidiabetic drugs. T-test and X² test were used to compare parameters between groups of CD and DM remission. A value of p<0.05 as considered to be statistically significant. Of the 108 CD patients (aged 39 years; 82,5% women; BMI 31,5 Kg/m²), 37% had DM diagnosis and 30% were treated with hypoglycemic agents and/or insulin. ACTH-producing pituitary adenoma was found in 80% of histopathological analyses. Hypertension and central obesity were found in most patients (100% and 97%, respectively). CD remission was achieved in 82.5% of the diabetic patients and 66% were also considered cured of DM after 1 year, showing no relationship between CD remission and DM cure (p=0.378). There was no statistically significant association between age, gender, BMI, lipid profile, 24h-UFC, FG, HbA1C at diagnosis and DM regression. Glucocorticoids dependence after TSS was also not associated with DM resolution (p=1.0). In this representative CD sample, prevalence of DM was consistent with literature, ranging from 30-50%. Short term CD remission does not seem to predict the resolution of diabetes, perhaps because metabolic effects persists even after correction of hypercortisolism and factors associated with DM resolution are heterogeneous.


Nothing to Disclose: CV, TML, SPG, AMDSM, FC, IMG, GC, TCR, MAC

27376 13.0000 FRI 406 A Is Cushing's Syndrome Remission Associated with Diabetes Regression? Analysis of a Retrospective Cohort of 108 Patients with Cushing's Disease 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Adrenal HPA Axis Friday, April 1st 3:15:00 PM FRI 394-408 7668 1:15:00 PM Glucocorticoid Actions (posters) Poster

Margreet A.E.M. Wagenmakers*1, Sean H.P.P. Roerink2, Tim H Schreuder3, Theo S Plantinga2, Suzanne Holewijn2, Dick H.J. Thijssen4, Johannes W.A. Smit1, Gerard A Rongen2, Alberto M. Pereira5, Anton J.M. Wagenmakers6, Romana T. Netea-Maier1 and Ad R.M.M. Hermus1
1Radboud University Medical Center, Nijmegen, Netherlands, 2Radboud University Medical Center Nijmegen, Netherlands, 3Radboud University Medical Center Nijmegen, Netherlands, 4Radboud University Medical Center, Nijmegen, Netherlands, 5Leiden University Medical Center, Leiden, Netherlands, 6Liverpool John Moores University, Liverpool, United Kingdom


Context: In active Cushing’s syndrome (CS), patients suffer from endothelial dysfunction and premature atherosclerosis. However, it is uncertain to what extent vascular health recovers after long-term remission. This is highly relevant as this relates to future development of cardiovascular disease.

Objective: To investigate whether micro- and macrovascular health is impaired after long-term remission of CS,  in patients with no or adequately treated co-morbidities compared to healthy control subjects 

Design and setting: Cross-sectional case–control study in two tertiary referral centers.

Patients and main outcome measures: 63 patients (remission of CS for ≥ 4 years) and 63 well matched controls were compared. In group A (58 patients and 58 controls) serum biomarkers associated with endothelial dysfunction, intima media thickness, pulse wave velocity and pulse wave analysis were studied. In group B (14 patients and 14 controls) endothelium dependent and independent vasodilation was studied in both conduit arteries (flow mediated dilation of brachial artery) and forearm skeletal muscle resistance arteries (vasodilator response to intra-arterial acetylcholine, sodium-nitroprusside and  NG-monomethyl-L-arginine using venous occlusion plethysmography).

Results There were no significant differences between the outcome measures of vascular health of patients and controls  in Group A and B.

Conclusion: In conclusion, vascular health of patients after long-term remission of Cushing’s syndrome seems to be comparable to that of healthy gender-, age and BMI matched controls, provided that the patients have no, or adequately controlled co-morbidities. Therefore, the effects of hypercortisolism per se on the vasculature may be reversible, which accentuates the need for stringent treatment of metabolic co-morbidities in these patients.



25324 14.0000 FRI 407 A Vascular Health in Patients in Long-Term Remission of Cushing's Syndrome and No or Adequately Treated Co-Morbidity Is Comparable to BMI-Matched Subjects 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Adrenal HPA Axis Friday, April 1st 3:15:00 PM FRI 394-408 7668 1:15:00 PM Glucocorticoid Actions (posters) Poster

Margreet A.E.M. Wagenmakers*1, Sean H.P.P. Roerink2, Dick H.J. Thijssen3, Maria T.E Hopman3, Anton J.M. Wagenmakers4, Alberto M. Pereira5, Johannes W.A. Smit1, Romana T. Netea-Maier1 and Ad R.M.M. Hermus1
1Radboud University Medical Center, Nijmegen, Netherlands, 2Radboud University Medical Center Nijmegen, Netherlands, 3Radboud University Medical Center, Nijmegen, Netherlands, 4Liverpool John Moores University, Liverpool, United Kingdom, 5Leiden University Medical Center, Leiden, Netherlands


Context: Although biochemical normalization and concomitant improvement of phenotype is achieved in most patients with Cushing’s syndrome (CS), centripetal fat distribution, physical disability and tiredness often persist after remission. Some of these factors may relate to impaired physical fitness in patients biochemically cured of CS, however this has never been investigated.

Objective: To investigate physical fitness level, as measured by peak oxygen uptake (VO2 peak) during a maximal exercise stress test and  to explore its relation with energy expenditure (EE), in patients in long-term remission of CS and matched controls.

Patients and methods: Patients in long-term (>4 years) remission of CS, who did not use any medication, except for thyroid hormone substitution,  were eligible. For each patient a sex-, estrogen status-, age-, BMI-, smoking-, ethnicity-, and physical activity level (measured via the METS-score) matched control subject was recruited from the general population. Physical fitness level was assessed using a maximal exercise stress test on a bicycle ergometer with an incremental exercise protocol. Daily EE was assed using an activity monitor (Sensewear Pro, worn for one week).

Main outcome measures: Peak oxygen uptake (VO2peak) during a maximal exercise stress test.

Results:  The patients in  long-term remission of CS (n=17) had a significantly lower peak oxygen uptake and maximal workload than the  matched controls (both P<0.05). Combined with a higher breath-by-breath minute ventilation (VE)/VO2 and a lower ventilatory threshold VO2in patients these results prompt the consideration of mitochondrial myopathy.  No significant differences were found in respiratory exchange ratio, peak heart rate and blood lactate-levels (all P>0.05).  Daily EE did not differ between the two groups.

Conclusions:  Patients in long-term remission of CS have a lower physical fitness level with indications of limitation in muscle respiratory chain function, despite similar daily EE, compared to well-matched healthy controls. The lower physical fitness level in patients is a  promising potential target for future (early and late) rehabilitation programs.



26227 15.0000 FRI 408 A Physical Fitness Level after Long Term Remission of Cushing's Syndrome: Does Biochemical Remission Lead to Full Functional Recovery? 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Adrenal HPA Axis Friday, April 1st 3:15:00 PM FRI 394-408 7668 1:15:00 PM Glucocorticoid Actions (posters) Poster

Yolanda M. Foster*1, Felicia Yan1, Zhe Zhang1 and Shana E. McCormack2
1Children's Hospital of Philadelphia, Philadelphia, PA, 2The Children's Hospital of Philadelphia, Philadelphia, PA


Background: In animal models, treatment with the hypothalamic neuropeptide oxytocin (OXT) leads to weight loss via increased energy expenditure and lipolysis.(1-3) OXT therefore has exciting translational potential for use in obesity.(4) However, the mechanistic basis of its effects on energy balance, in particular outside the CNS, is not well-characterized.

Objective: To examine the association between OXT and mitochondrial oxidative phosphorylation (OXPHOS) (5) in peripheral energy-requiring tissues using a two-pronged approach, bioinformatics and in vitro measurements.

Methods: Bioinformatics. NCBI/GEO gene expression profiles were queried using the locally developed Awsomics toolbox. A curated collection of 35 datasets from experiments including pharmacologic or genetic disruption of the mitochondrial respiratory chain (RC) was used to identify conditions in which OXT was differentially expressed, using ANOVA with Bonferroni correction. Tissue culture. The effects of physiologic OXT (10uM x 24 hours) (6) on mitochondrial parameters was assessed using Fluorescence-Activated Cell Sorting Analysis (9) in liver-like hepatoma (HepG2) and muscle-like rhabdomyosarcoma (RD) cells under different nutrient conditions (5 mM versus 25 mM glucose).

Results: Bioinformatics. In 7/35 datasets, OXT showed differential expression. OXT was decreased 39% in skeletal muscle from humans with a primary mitochondrial RC diseases, and also decreased in 2 skin fibroblast cell lines from humans with mitochondrial RC diseases (specifically, coenzyme Q10 deficiency and mitochondrial ATP synthase deficiency), 60% and 57%, respectively. In contrast, in human neuroblastoma cells treated for ~1 week with rotenone (a mitochondrial RC complex I inhibitor), liver from mice with liver-specific Pdss2 deletion and associated coenzyme Q9 deficiency, and in cochlea from mice with mtDNA mutations, a marked increase in OXT expression was observed (54%, 224%, and 145%, respectively). Tissue Culture. In RD muscle-like cells, OXT produced a 45% ± 24% (SEM) increase in mitochondrial content in 5 mM glucose; in 25 mM glucose, 24% ± 41% (SEM). In HepG2 liver-like cells, OXT produced a 40% ± 25% (SEM) increase in mitochondrial content in 5 mM glucose; in 25 mM glucose, 8% ± 7% (SEM). Accounting for content, mitochondrial oxidant burden was decreased in RD cells (72% and 75% in 5 and 25 mM glucose, respectively) and also in HepG2 cells (83% and 84% in 5 and 25 mM glucose, respectively).

Conclusions: OXT expression is often altered in response to OXPHOS disruption in peripheral tissues; as in previous studies (7), the direction of effect is tissue-specific and may be compensatory. Indeed, short-term treatment of energy-modulating cell types (liver, muscle) in vitro with OXT increased mitochondrial content, which could be the basis of increased energy expenditure. Follow-up mechanistic studies are ongoing.


Nothing to Disclose: YMF, FY, ZZ, SEM

25740 3.0000 FRI 478 A Tissue-Specific Effects of Oxytocin on Mitochondrial Bioenergetics 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 3:15:00 PM FRI 476-494 7669 1:15:00 PM Neuroendocrinology (posters) Poster

Haluk Kelestimur*, Gokcen Ozdemir, Ozgur Bulmus, Sinan Canpolat and Mete Ozcan
Firat University Medical School, Elazig, Turkey


Although the neurotransmitters, dopamine, norepinephrine and serotonin, are suggested to be related to the control of reproductive axis, underlying mechanisms have not been well known. Two novel hypothalamic peptides, kisspeptin and RFRP-3, may be important regulators of the reproductive axis. Kisspeptin is currently recognized as the most potent activator of the hypothalamus-pituitary-gonadal axis, and RFRP-3, an orthologous mammalian peptides of gonadotropin-inhibitory hormone, is suggested to be an inhibitor of the axis. These neurotransmitters may modulate the reproductive axis by means of these peptides. Therefore, in the current study, the effects of these neurotransmitters on kisspeptin and RFRP-3 neurons have been investigated. The rHypoE-7 and rHypoE-8 cell lines expressing kisspeptin and RFRP-3 were used as models to explore the effects of Arg-Phe-amide-related peptides on the reproductive axis. We investigated the effects of the neurotransmitters on [Ca2+]i and release of kisspeptin and RFRP-3 in these immortalized neurons. The changes in [Ca2+]i in these neurons were investigated by using in vitro calcium imaging system. rHypoE-7 and rHypoE-8 cells were placed on glass coverslip and loaded with 1 μM Fura-2 AM. [Ca2+]i responses were quantified by the changes in 340/380 ratio. The peptides released into the medium were detected via enzyme-linked immunosorbent assay. None of the neurotransmitters caused a significant change in [Ca2+]i. Only serotonin (100 nM) significantly increased kisspeptin secretion in rHypoE-7 cells (at 90 minutes of exposure). Therefore, serotonin may exert its effect on the reproductive axis by means of modulating kisspeptin secretion.


Nothing to Disclose: HK, GO, OB, SC, MO

24404 4.0000 FRI 479 A Serotonin Increases Kisspeptin Secretion in Immortalized Hypothalamic RF-Amide-Related Peptide Neurons 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 3:15:00 PM FRI 476-494 7669 1:15:00 PM Neuroendocrinology (posters) Poster

Nicole H Bellefontaine*1, Margaret B Allison2, Martin Grosvenor Myers Jr.3, David Garcia Galiano1 and Carol F Elias1
1University of Michigan, Ann Arbor, MI, 2University of Michigan, 3University of Michigan Medical School, Ann Arbor, MI


The ventral premammilary nucleus (PMv) of the hypothalamus is a putative sexually dimorphic nucleus that is responsive to metabolic signals, opposite sex odorant cues, and expresses sex steroid receptors. Projections originating from the PMv are dense within the hypothalamus and are found largely in areas implicated in the control of reproductive physiology. Therefore, the PMv is well positioned to integrate metabolic and sex steroid cues to signal to the neuroendocrine reproductive axis. Lesions to the PMv disrupts estrous cyclicity and alters Kiss-1 and GnRH mRNA. Excitotoxic lesions to the PMv also impairs the ability of exogenous leptin administration to rescue cyclicity during an acute fast. However, the cell populations within the PMv that control these facets of reproductive physiology remains unclear. In this study we describe a novel cell population within the PMv expressing the dopamine transporter (DAT). To examine the DAT population and potential sexual dimorphism, we crossed a DAT-Cre to a tdTomato reporter line. There was no difference in the number of tdTomato positive cells between male and female adult mice. Using DAT-Cre mice, we injected a viral tracer that expresses a synaptophysin-mCherry fusion protein in a cre recombinase-dependent manner to determine projection sites of the PMv DAT cell population. DAT cells from the PMv project across the hypothalamus, including the arcuate nucleus (ARH), ventromedial hypothalamus, and the medial preoptic region. There appeared to be more dense projections in males compared to females, suggesting a potential sexual dimorphism in the projections of the DAT PMv population. Interestingly, the DAT cells in the PMv do not coexpress tyrosine hydroxylase (TH), however these cells project to TH-positive cells in the ARH. To examine whether the DAT cell are responsive to leptin, DAT-Cre tdTomato mice were injected with exogenous leptin. Immunohistochemical analyses for leptin-induced P-STAT3 revealed that 35% of tdTomato-positive cells colocalize with P-STAT3-ir. DAT cells within the PMv are also responsive to changes in sex steroid milieu. DAT mRNA is decreased in ovariectomized (OVX) mice when compared to intact diestrus female mice. Estradiol replacement to OVX mice (OVX + E) restores DAT mRNA levels to diestrus levels, suggesting that DAT expression is sensitive to sex steroid levels. Here we have begun to characterize a novel population within the PMv that express DAT. PMv DAT cells are a direct target for the metabolic hormone leptin and levels of DAT mRNA are altered by sex steroids. Further, PMv DAT cells project across the hypothalamus, including areas involved in reproductive control. Therefore, the PMv DAT cell population may play an important role in integrating metabolic and sex steroid signals to the central reproductive axis. Future studies are needed to further define the physiological role of the PMv DAT expressing cells.


Nothing to Disclose: NHB, MBA, MGM Jr., DG, CFE

27499 5.0000 FRI 480 A DAT Neurons in the Pmv Are Responsive to Metabolic and Sex Steroid Cues 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 3:15:00 PM FRI 476-494 7669 1:15:00 PM Neuroendocrinology (posters) Poster

Erin Semple*1, Shokufeh Nourollahi2 and Jennifer Wootton Hill3
1University of Toledo, Holland, OH, 2University of Toledo, Toledo, OH, 3University of Toledo School of Medicine, Toledo, OH


Sexual dysfunction and reduced fertility have been found to be associated with metabolic disorders such as metabolic syndrome, diabetes, and PCOS. Understanding where these regulatory functions diverge and where they run in parallel should enhance our ability to address metabolic diseases and sexual disorders separately as well as when the two are comorbid. The melanocortin pathway has been implicated in both metabolism and reproduction. When the melanocortin 4 receptor (MC4R) is knocked out globally, male mice have an obese, diabetic phenotype and also show erectile dysfunction. We hypothesize that the paraventricular nucleus of the hypothalamus (PVN) may be a key site of melanocortin-mediated regulation of both sexual behavior and metabolism. To test this hypothesis, the metabolic phenotype and sexual behavior of a mouse model in which MC4R is expressed only on neurons in the PVN was compared to MC4R null mice. The role of this pathway in metabolism and sexual behavior was explored in both male and female mice. To test sexual function, both copulatory behaviors and fertility were assessed. Mice were paired with a sexually experienced mouse of the opposite sex and their behavior was filmed between 8pm and 2am. The behavioral videos were scored based on male sexual behaviors such as mounting, intromission, and ejaculation, while the females were scored based on lordosis, solicitations, and rejection behaviors. Fertility was assessed through successful births, sex hormone concentrations, and gonadal histology. A metabolic profile was obtained from these mice through the use of GTT, NMR, and weight gain. Expression of MC4R in the PVN attenuated the metabolic deficits seen in MC4R null mice but did not completely reverse the phenotype to that of wild-type controls. This study sheds light on the role of MC4R in the PVN in the neurocircuitry underlying both metabolism and sexual behavior.


Nothing to Disclose: ES, SN, JWH

25798 6.0000 FRI 481 A Melanocortin 4 Receptors in the Paraventricular Nucleus of the Hypothalamus Influence Metabolism and Sexual Behavior in Transgenic Mice 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 3:15:00 PM FRI 476-494 7669 1:15:00 PM Neuroendocrinology (posters) Poster

Raul M. Luque*1, André Sarmento-Cabral2, Lisa C. Halliday3, Maria M Malagon2, Justo Pastor Castano2 and Rhonda D. Kineman4
1Instituto Maimónides de Investigación Biomédica de Córdoba (IMIBIC)/Hospital Universitario Reina Sofia, CIBER de la Fisiopatología de la Obesidad y Nutrición (CIBERobn) and ceiA3, Córdoba, Spain, 2University of Cordoba; Instituto Maimónides de Investigación Biomédica de Córdoba (IMIBIC)/Hospital Universitario Reina Sofia, CIBER de la Fisiopatología de la Obesidad y Nutrición (CIBERobn) and ceiA3, Córdoba, Spain, 3University of Illinois at Chicago, Chicago, IL, 4& Medicine, Section of Endocrinology, Diabetes and Metabolism, University of Illinois at Chicago, Chicago, IL


Adipose tissue represents a true endocrine organ that dynamically secretes multiple hormones referred to as adipokines that regulate key physiological processes (i.e. metabolism, inflammation, reproduction, etc.). Adipokines (i.e. leptin, resistin, adiponectin) and their receptors are also expressed and regulated in other endocrine tissues, including the pituitary. Therefore, locally-produced adipokines, as well as those derived from adipose tissue, might comprise a relevant regulatory circuit to modulate pituitary function. However, the direct actions of different adipokines in the pituitary remain controversial, and previous studies in normal pituitary cells are mostly limited to nonprimate species. Here, primary pituitary cell cultures from two normal nonhuman primates species [Papio anubis (n=7) and Macaca fascicularis (n=3)] were used as model systems to determine the direct impact of key adipokines (leptin, resistin, and adiponectin; 4h-incubation) on pituitary cell function. In these models, both leptin (10ng/ml) and resistin (0.1nM) stimulated GH release, a response that was blocked by somatostatin. Interestingly, leptin, but not resistin, significantly increased the expression of the transcription factor Pit-1. Conversely, adiponectin (10nM) decreased basal GH release, and was also able to inhibit GHRH-, but not ghrelin-stimulated GH secretion, independent of changes in Pit-1. Use of inhibitors specifically blocking different signaling pathways revealed that these adipokines activate both common (AC/PKA and PI3K) and distinct (PLC/PKC, intra-/extra-cellular calcium, MAPK or mTOR) signaling pathways to exert their effects on GH secretion. Of note, these adipokines not only regulated somatotrope function but also controlled other pituitary cell types. Specifically: 1) Leptin stimulated PRL/ACTH/FSH but not LH/TSH release; 2) adiponectin stimulated PRL, inhibited ACTH and did not alter LH/FSH/TSH release; and 3) resistin increased ACTH release and did not alter PRL/LH/FSH/TSH secretion, all these effects being mediated through activation of specific signaling cascades. Moreover, treatment with these three adipokines directly regulated the expression of key receptors known to control pituitary cell function (e.g. receptors for GHRH, ghrelin, somatostatin, dopamine, CRF, Kiss1, insulin and/or IGF-I). Taken together, these results show for the first time that leptin, adiponectin and resistin can directly modulate the function of different pituitary cell types in two primate models, by regulating hormone release through common and distinct intracellular signaling pathways, as well as by regulating the expression of receptors and/or transcription factors important in the normal function of all the pituitary cell types.


Nothing to Disclose: RML, AS, LCH, MMM, JPC, RDK

25900 7.0000 FRI 482 A Adipokines (leptin, adiponectin and resistin) Differentially Regulate All Hormonal Cell Types in Primary Pituitary Cell Cultures from Two Primate Species (Papio anubis and Macaca fascicularis) 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 3:15:00 PM FRI 476-494 7669 1:15:00 PM Neuroendocrinology (posters) Poster

Peyton Wood Weems*1, Robert L. Goodman2, Lique M Coolen1 and Michael N Lehman1
1University of Mississippi Medical Center, Jackson, MS, 2West Virginia University School of Medicine, Morgantown, WV


Dynorphin (Dyn) is an endogenous opioid peptide that plays an important role in mediating the negative feedback influence of progesterone on pulsatile GnRH secretion in sheep, as well as the control of energy balance and food intake. Dyn action is mediated via its high affinity receptor, kappa opioid receptor (KOR). Although the distribution of KOR has previously been described in rodents, until recently it had yet to be shown in sheep. Recently we reported the presence KOR in kisspeptin/NKB/dynorphin (KNDy) neurons of the sheep and rat arcuate nucleus (ARC), consistent with its role in the control of GnRH pulses. However, KNDy neurons only accounted for ~43% of KOR-ir cells within the ovine ARC. Thus, Dyn from KNDy or other neurons may also act upon adjacent non-KNDy, KOR-containing neurons in the ARC to regulate GnRH pulse frequency. To determine the phenotype of non-KNDy, KOR-containing neurons in the sheep ARC, we analyzed three populations previously shown to be regulated by Dyn: pro-opiomelanocortin (POMC), agouti-related peptide (AgRP), and tyrosine hydroxylase (TH)-positive, tuberoinfundibular dopamine neurons.   In addition, we examined co-localization of KOR in neurons containing the NKB receptor, NK3R, since NK3R are present in approximately 50% of KNDy neurons but also in a sizable population on non-KNDy cells.  Tissue sections from ewes perfused during the luteal phase of the estrous cycle (n = 4) were processed for dual-label immunofluorescent detection of KOR and POMC, AgRP, TH, or NK3R. Confocal analysis of single and double-labeled cells revealed that KOR was co-localized in 45% of POMC, 59% of AgRP, 42% of TH neurons and 81% of NK3R cells. Co-localization of KOR in POMC, AgRP and TH arcuate cells are consistent with previous pharmacological findings suggesting regulation of food intake and prolactin secretion via Dyn, but in addition raise the possibility that Dyn actions upon POMC, AgRP, TH cells may contribute to its regulation of GnRH pulse frequency. In addition, non-KNDy, NK3R-containing cells may serve as an additional substrate by which NKB and dynorphin act as start and stop signals, respectively, to control individual GnRH pulses.


Nothing to Disclose: PWW, RLG, LMC, MNL

25963 9.0000 FRI 484 A Phenotypic Identification of Kappa Opioid Receptor-Containing Neurons in the Ovine Arcuate Nucleus 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 3:15:00 PM FRI 476-494 7669 1:15:00 PM Neuroendocrinology (posters) Poster

Manqi Wang* and Matthew D. Whim
LSU Health Sciences Center, New Orleans, LA


Hypoglycemia is a serious condition that can lead to loss of consciousness and is potentially fatal. The effects of hypoglycemia are prevented during fasting by the activation of the counter-regulatory response (CRR). Epinephrine secreted from adrenal chromaffin cells contributes to the CRR by increasing hepatic glucose production but how epinephrine secretion is regulated during fasting remains unclear. To address this question we studied the functioning of the sympatho-adrenal system in mice that were fed ad lib or fasted for 1 day. As previously reported, urine epinephrine levels were significantly higher after fasting. Blood glucose levels did not differ significantly between fed and fasted littermates, indicating that the CRR could maintain euglycemia even after a substantial period of fasting. Because neuropeptide Y (NPY) is a transmitter that is co-released with epinephrine from chromaffin cells we repeated these experiments in NPY knockout (NPY k/o) mice. In these animals fasting did not increase the urine levels of epinephrine and the mice were hypoglycemic. To determine how the loss of NPY prevented epinephrine release we assessed both pre- and post-synaptic signaling at the preganglionic → chromaffin cell synapse. Post-synaptically, catecholamine release was evoked from chromaffin cells in vitro by a train of voltage clamp depolarizations and monitored using carbon fiber amperometry. There was no difference in the amplitude of amperometric events between fed and fasted wild type (wt) mice. However fasting significantly increased the amplitude of amperometric events in NPY k/o mice, indicating that the catecholamine secretory capacity from isolated cells was negatively regulated by NPY. Because this could not explain the observed decrease in epinephrine release in vivo we next considered whether a presynaptic mechanism was involved. Using acute adrenal slices we found that food deprivation was associated with an increase in the amplitude of the evoked EPSC monitored in chromaffin cells from wt mice. In contrast, the amplitude of the evoked EPSC was reduced in the fasted NPY k/o animals compared to fed littermates. Food deprivation led to a decrease in the paired-pulse ratio (PPR) in wt animals, but to an increase in the PPR in NPY k/o mice, consistent with the involvement of a presynaptic component. Thus food deprivation is associated with an NPY-dependent plasticity at the preganglionic → chromaffin cell synapse. Furthermore, pharmacological inhibition of Y5 receptors blocked the fasting-induced change in synaptic plasticity and epinephrine release, resulting in hypoglycemia. We conclude that NPY, likely secreted from chromaffin cells, increases pre-ganglionic → chromaffin cell synaptic efficacy and this contributes to the maintenance of euglycemia during fasting.


Nothing to Disclose: MW, MDW

26234 10.0000 FRI 485 A Sympathetic Synaptic Plasticity Contributes to the Maintenance of Euglycemia during Fasting 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 3:15:00 PM FRI 476-494 7669 1:15:00 PM Neuroendocrinology (posters) Poster

Hiranya Pintana*, Pongpan Tanajak, Wasana Pratchayasakul, Piangkwan Sa-nguanmoo, Titikorn Chunchai, Pattarapong Satjaritanun, Linlada Leelarphat, Nipon Chattipakorn and Siriporn C Chattipakorn
Chiang Mai University, Chiang Mai, Thailand


Chronic high fat diet consumption has been shown to lead to obese-insulin resistance.  Interventions including calories restriction (CR) and antidiabetic drug dipeptidyl peptidase-4 (DDP-4) inhibitor have been shown to be beneficial in obese-insulin resistant subjects (1-3).  However, previous studies demonstrated that obese-insulin resistant condition not only causes metabolic disturbance, but also leads to brain insulin resistance and impaired cognitive function (4,5).  Nevertheless, the effects of CR and CR with DDP-4 inhibitor as well as the comparative efficacy of these interventions on brain insulin sensitivity, brain mitochondrial function, hippocampal synaptic plasticity and cognition in obese-insulin resistant condition have never been investigated.  Therefore, the present study determined whether CR or CR with DPP-4 inhibitor exert similar efficacy for neuroprotection in obese-insulin resistant rats.  We tested the hypothesis that CR and CR plus DPP-4 inhibitor improves cognition in obese-insulin resistant rats by restoring brain insulin sensitivity, brain mitochondrial function and hippocampal synaptic plasticity.  Twenty-four male Wistar rats were divided into 2 groups and fed with either a normal diet (ND) or a high-fat diet (HFD) for 12 weeks.  At week 13, HFD-fed rats were divided into 3 subgroups (n=6/subgroup) to receive one of the following treatment: vehicle, CR (60% of energy received during previous 12 weeks) with vehicle, or CR plus DPP-4 inhibitor (vildagliptin (Vil) 3 mg/kg/day, p.o.) for 4 weeks.  At the end of treatments, cognitive function, metabolic parameters, brain insulin sensitivity, hippocampal synaptic plasticity and brain mitochondrial function were determined in each rat.  The results showed that HFD-fed rats with vehicle demonstrated weight gain with peripheral insulin resistance, dyslipidemia, oxidative stress, brain insulin resistance, impaired brain mitochondrial function and cognitive dysfunction, when compared with ND-fed rats.  Both CR and CR with Vil restored oral glucose tolerance test (4.8±0.3 mg/dlxminx104 in CR group; 4.9±0.2 mg/dlxminx104 in CR with Vil), compared to vehicle-treated HFD-fed rats (6.0±0.3 mg/dlxminx104; p<0.05) and improved lipid profiles in HFD-fed rats (p<0.05).  However, only CR with Vil, significantly restored brain insulin sensitivity, brain mitochondrial function, hippocampal synaptic plasticity and cognitive function.  These findings indicate that although calories restriction can restore the metabolic profiles under obese condition, it cannot improve brain function impaired by obese-insulin resistant condition.  Combined CR with DPP-4 inhibitor provides the protective effects on brain function in obese-insulin resistant condition.


Nothing to Disclose: HP, PT, WP, PS, TC, PS, LL, NC, SCC

24266 11.0000 FRI 486 A Dipeptidyl Peptidase-4 (DPP-4) Inhibitor Exerts Better Neuroprotection Than Calories Restriction By Attenuating Mitochondrial Dysfunction Impaired By Obese-Insulin Resistance in Rats 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 3:15:00 PM FRI 476-494 7669 1:15:00 PM Neuroendocrinology (posters) Poster

Ari Loren Mendell*1, Sarah Atwi2, Craig D.C. Bailey1, Dan McCloskey3, Helen Edith Scharfman4 and Neil James MacLusky5
1University of Guelph, 2University of Toronto, 3City University of New York, 4Nathan Kline Institute for Psychiatric Research, Orangeburg, NY, 5University of Guelph, Guelph, ON, Canada


Androgen loss is an important clinical concern because of its cognitive and behavioral effects. Changes in androgen levels are also suspected to contribute to psychiatric illness. However, the available data on the effects of androgen deprivation on areas of the brain that are central to cognition, including the hippocampus, are mixed. A method that has been widely employed to investigate changes in the hippocampus in response to gonadal steroids, morphological analysis of pyramidal cells, was used to investigate if structural changes could potentially explain the mixed cognitive effects that have been observed after androgen loss in males, particularly following androgen ablation therapy. In this study, male Sprague-Dawley rats were orchidectomized or sham-operated. Two months later, their brains were Golgi-impregnated for morphological analysis. Morphological endpoints were studied in areas CA3 and CA1, with the corresponding measures performed in females for comparison. Intact females sacrificed at proestrus and metestrus, as well as two months following ovariectomy, were included for analysis. There were robust enhancements of mossy fiber afferents and dendritic arborisation of pyramidal neurons in the CA3 area of the hippocampus of orchidectomized males, but not in CA1. Remarkably, dendritic length of CA3 pyramidal cells greatly increased, while spine density slightly declined. These effects stood in stark contrast from those in females, in which overall dendritic structure was minimally affected by ovariectomy, although dendritic spine density was greatly reduced in both the CA3 and CA1 hippocampal areas. Sex differences and subfield-specific effects of gonadal hormone deprivation on the hippocampal circuitry may help to explain diverse behavioral effects reported in males and females after gonadectomy, as well as other conditions associated with declining gonadal hormone secretion.


Nothing to Disclose: ALM, SA, CDCB, DM, HES, NJM

26247 12.0000 FRI 487 A Dramatic Extension of CA3 Apical Dendritic Length Accompanies Mossy Fiber Expansion and Spine Loss in Orchidectomized Male Rats 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 3:15:00 PM FRI 476-494 7669 1:15:00 PM Neuroendocrinology (posters) Poster

Meghan Ferguson*1, James McTague1, Constance L Chik2 and Anthony-K Ho2
1University of Alberta, Edmonton, AB, Canada, 2Univ of Alberta, Edmonton, AB, Canada


Phosphorylation plays an important role in determining the intracellular distribution and transcription co-activation activity of CREB-regulated transcription coactivators (CRTCs). Whether the equivalent phosphorylation sites on CRTC1 and CRTC2 perform the same function in rat pinealocytes remains unclear. Three specific phosphorylation sites (S151, S245 and S 277 on CRTC1 and the equivalent S171, S274 and S306 on CRTC2) were investigated by over expressing their phosphorylation defective mutants in the rat pinealocyte. Whereas norepinephrine (NE) stimulation can cause additional dephosphorylation of sites besides the three mutated, for nuclear localization, only dephosphorylation of S151 and S245 of CRTC1 (or S171 and S274 of CRTC2) is sufficient to reproduce the effect of NE. As for nuclear exit following adrenergic blockade, our results suggest that whereas rephosphorylation of S151, S245 and S277 contributes to the nuclear exit o f CRTC1, only rephosphorylation of S171 appears to be required for the nuclear exit of CRTC2. When assessing their co-activation activities, all CRTC1 and CRTC2 mutants show similar enhancing effects on the NE-stimulated arylalkyl-N-acetyltransferase (Aanat) transcription.  However, in the absence of NE stimulation, only CRTC1 mutants with the double or triple mutations, involving S151, S245 and S277, can significantly elevate Aanat transcription.  CRTC2 with equivalent mutations are without effects, in spite of their presence in the nucleus.  This stimulation by CRTC1 is also observed in selected, but not all, CREB-targeted genes. Together, we show that, between CRTC1 and CRTC2, whereas the entry into the nucleus involves dephosphorylation of equivalent sites, there are significant differences in the rephosphorylation requirement for nuclear exit. In addition, whereas nuclear entry of CRTC1 alone appears to be sufficient to stimulate Aanat transcription, this is not the case for CRTC2.


Nothing to Disclose: MF, JM, CLC, AKH

26369 13.0000 FRI 489 A Role of Specific Phosphorylation Sites on CRTC1 and CRTC2 in the Nuclear Localization and Nuclear Exit in Rat Pinealocytes 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 3:15:00 PM FRI 476-494 7669 1:15:00 PM Neuroendocrinology (posters) Poster

Guangfu Hu, Mu-Lan He, Wendy K.W. Ko and Anderson O L Wong*
University of Hong Kong, Hong Kong, China


In mammals, the gene for tachykinin 1 (TAC1) encodes two mature peptides, namely substance P (SP) and neurokinin A (NKA), which preferentially bind with type 1 (NK1R) and type 2 neurokinin (NK) receptors (NK2R), respectively, but with low affinity for type 3 NK receptor (NK3R).  To date, little is known regarding the biological actions of TAC1 gene products in lower vertebrates, especially in fish species.  Using grass carp as animal model, the direct effects of SP and NKA on pituitary hormone regulation were tested at the pituitary level in bony fish.  As a first step, TAC1, NK1R, NK2R and NK3R cDNA were pulled out from grass carp pituitary, sequence analysis reveals that carp TAC1 also encodes the mature peptides for SP and NKA and the carp NK1R, NK2R and NK3R are highly homologous to their human counterparts with the typical structures of G protein-coupled receptor.  Expression of these newly cloned NK receptors in HEK293 cells also confirmed that they were functionally coupled with cAMP-, PKC- and Ca2+-dependent signaling cascades.  In these expression studies, carp SP was found to have the highest efficacy and potency for NK1R activation when compared with NKA and TAC3 gene products (including NKB and NKBRP) of carp origin.  Although not effective at low doses, high doses of carp SP and NKA were also effective in triggering NK3R activation.  Parallel experiments on NK2R expression, interestingly, revealed that the receptor did not differentiate TAC1 and TAC3 gene products but could be activated by SP, NKA, NKB and NKBRP with comparable efficacy/potency.  In grass carp pituitary cells, SP and NKA of carp origin could induce luteinizing hormone (LH), prolactin (PRL) and somatolactin α (SLα) release with parallel rises in PRL and SLα mRNA expression.  Of note, short-term exposure to SP induced LH secretion but prolonged treatment with SP could lead to a mild reduction in LHβ mRNA expression.  Using subtype-specific antagonists for the three types of NK receptors, the stimulatory effects on LH, PRL and SLα were shown to be mediated by NK1R, NK2R, and NK3R, respectively.  By pharmacological blockade of respective signaling targets, SP and NKA stimulation on hormone release for LH and SLα and gene expression for SLα were confirmed to be the results of AC/cAMP/PKA, PLC/ IP3/PKC and Ca2+/CaM/CaMK-II signal activation.  The signaling mechanisms for PRL responses were similar, except that the PKC component was not involved.  Regarding the inhibitory effect on LHβ mRNA expression induced by prolonged treatment with SP, the cAMP/PKA and PLC/PKC pathways were involved but not the Ca2+-dependent cascades.  Taken together, our results suggest that TAC1 gene products, including SP and NKA, can play a role in LH, PRL and SLα regulation in the carp pituitary by overlapping post-receptor signaling mechanisms coupled to NK1R, NK2R and NK3R, respectively.


Nothing to Disclose: GH, MLH, WKWK, AOLW

26504 14.0000 FRI 490 A Regulation of Luteinizing Hormone, Prolactin and Somatolactin Secretion and Gene Expression By TAC1 Gene Products in Carp Pituitary Cells: Signal Transduction and Receptor Specificity at Pituitary Level 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 3:15:00 PM FRI 476-494 7669 1:15:00 PM Neuroendocrinology (posters) Poster

Julie Brossaud*, Hélène Roumes, Aloïs Lemelletier, Marie-Pierre Moisan, Véronique Pallet, Anabelle Redonnet and Jean-Benoit Corcuff
Nutrition et Neurobiologie Intégrée laboratory, Bordeaux, France


Vitamin A metabolite retinoic acid (RA) plays a major role in the aging adult brain plasticity. Conversely, chronic excess of glucocorticoids (GC) elicits some deleterious effects in the hippocampus. As synaptic plasticity depends on mechanisms related to cell morphology, we questioned the involvement of RA and GC in the remodelling of actin cytoskeleton of neurons. We investigated some pathways of neuroplasticity upon administration of RA and GC pathways agonists in the hippocampal HT22 cell line on the expression of CaMKII and Arc genes, and F-actin cytoskeleton organization.

Cell morphology and actin cytoskeleton, and plasticity genes expression changed dramatically upon RA and/or dexamethasone (Dex, a GC agonist) treatments. RA increased and Dex decreased CaMKII mRNA expression, and RA increased and Dex did not modify Arc mRNA expression. Actin expression and abundance where unchanged by RA and/or Dex. Conversely, F-actin organization was dramatically modified by both RA and Dex. RA induced actin structures favouring neural plasticity whereas Dex actions were the opposite. RA treatment increased calpain activity whereas Dex decreased of calpain activity.

The interaction between the RA and Dex signalling pathways on plasticity genes expression and actin cytoskeleton mirrors prior results shown in these cells on other genes including BDNF. The RA and Dex interacting effects could thus be elicited by a direct action of RA and Dex or via autocrine BDNF action. Pharmacological targeting common elements of the retinoid, GC and BDNF pathways e.g. calpains may counteract some of the deleterious effects of GCs and of the age-related cognitive impairment.


Nothing to Disclose: JB, HR, AL, MPM, VP, AR, JBC

26952 15.0000 FRI 491 A Retinoids and Glucocorticoids Target Actin Cytoskeleton Remodelling in Hippocampal HT22 Cells 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 3:15:00 PM FRI 476-494 7669 1:15:00 PM Neuroendocrinology (posters) Poster

Julie Brossaud*1, Emilie Leprovost1, Nina Franzoni1, Dominique Ducint2 and Jean-Benoit Corcuff1
1Nutrition et Neurobiologie Intégrée laboratory, Bordeaux, France, 2University hospital of Bordeaux, Bordeaux, France


Glucocorticoids (GC) are secreted in response to the perception of the homeostasis perturbation, a.k.a stress. Some pathophysiological situations are known to be associated with chronic and deleterious action of the GC, particularly in the central nervous system. Their intracellular availability is dependent, at least on part, on the intra-cellular activity of 2 enzymes: 11β-hydroxysteroid-deshydrogenase type 1 and 2 (HSD1 and HSD2) which transform active GC (cortisol or corticosterone) in inactive GC (cortisone or deshydrocorticosterone) for HSD1 and vice versa for HSD2.

The aim of this work was to develop a method to investigate the modifications of the activity of the 2 enzymes in 2 cell lines (HT22, hippocampic and BV2, glial). Previous method used cell lysates to examine enzymes functions. However, this method rested upon the use on tritiated GC and on coercing the enzymes to function in a contraflow, non physiological, direction.

As dexamethasone (Dex) is known to increase HSD1 and HSD2 expressions, we investigated the effects of a 96h-Dex treatment (10 -6M) on the expression (qPCR) and activity of HSD1 and 2 in these cell lines. The enzymatic activities were accessed by the evaluation of the conversion rate of the spiked corticosterone (C) or deshydrocorticosterone (DHC) in the supernatant of the cells by mass spectrometric assay. Briefly, HT22 or BV2 cells were incubated during 24h with either C or DHC (25ng/mL). Then, GC in the supernatant were extracted with dichloromethane. After evaporation, the dried extract was dissolved with mobile phase, 25 µL were injected in a LC-MSMS system (QTrap, ABsciex). Specific transitions allowed the quantification of C and DHC. Thus, the synthesis of C and DHC in the supernatant of cells subjected to DHC or C only allowed the evaluation of HSD1 and HSD2 relative activities.

In HT22 cells, there was no significant expression of HSD1 with or without Dex treatment. The consequence on enzymatic activity concurred: there was no conversion of DHC to C. While HSD2 was weakly expressed in these cells, Dex treatment significantly increased HSD2 expression. However, the enzymatic activity was moderate in basal condition and did not increase significantly with Dex treatment. In BV2 cells, both enzymes were basally expressed. The expression of HSD1 was significantly increased by Dex but unchanged for HSD2. However, the HSD1 enzymatic activity was predominant without treatment (about 90 % of conversion from C to DHC) and failed to increase with Dex. In contrast, basal activity of HSD2 (about 7%) increased with Dex treatment.

In conclusion, this work underlines the utility to investigate enzymatic activity from living cells as


Nothing to Disclose: JB, EL, NF, DD, JBC

26982 16.0000 FRI 492 A 11 β-Hydroxysteroid-Deshydrogenase Activity in HT22 Neuronal and BV2 Glial Cell Lines 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 3:15:00 PM FRI 476-494 7669 1:15:00 PM Neuroendocrinology (posters) Poster

June Zhou1, Mark Burns2, Richard Amdur3, Juan M Saavedra2 and Marc R. Blackman*4
1Washington DC VA Medical Center, Washington, DC, 2Georgetown University School of Medicine, Washington, DC, 3George Washington University School of Medicine, Washington, DC, 4Washington DC VAMC, Washington, DC


Introduction: Glucose and its intermediate metabolite, lactate, are essential fuels for maintenance of brain cell function. Patients with traumatic brain injury (TBI) exhibit immediate increases followed by prolonged decreases in brain glucose metabolism, as assessed by PET brain imaging. Understanding the molecular pathways of brain glucose utilization following TBI is vital for developing effective therapeutic strategies for this condition.  In this study, we measured cortical and hippocampal expression of genes related to glucose metabolism in a mouse model of TBI.

Hypothesis: TBI is associated with time dependent alterations in gene expression of glucose utilization-related enzymes and transporters in specific brain regions.

Methods: Adult male C57BL/6J mice (n= 6/group) were exposed to controlled cortical impact injury of the left cortex versus sham injury (control). Cortex and hippocampus ipsilateral and contralateral to the injury were collected after 6 hours, 1, 3, 7, 14, 21, and 28 days.  mRNA’s were measured by qRT-PCR for: (1) three key enzymes in glucose metabolism, HK1 (Hexokinase 1), PKm (Pyruvate kinase), and Pdhb (Pyruvate dehydrogenase); (2) two glucose transporters (Glut-1 and Glut-3), (3) two lactate transporters (MCT1 and MCT2); (4) two genes that closely link glucose metabolism with apoptosis: HK2, (an isoform of Hexokinase) and GPR81 (lactate receptor). We used t-tests to compare TBI versus control data at individual time points, considering p<0.05 as significant.

Results: (1) mRNA levels of HK1, PKm, and Pdhb all increased 6 hours after injury in the contralateral cortex, followed by decreases at subsequent time points in the ipsilateral cortex and hippocampus.  (2) Capillary glucose transporter Glut-1 mRNA increased at the different  time points in the ipsilateral cortex and hippocampus, reflecting increased glucose demand. In comparison, neuronal glucose transporter Glut-3 mRNA decreased at various time points in the ipsilateral cortex and hippocampus, reflecting diminished glucose transport into neurons.  (3) Astrocyte lactate transporter MCT-1 mRNA increased, whereas neuronal lactate transporter MCT-2 deceased, reflecting diminished  lactate transport into neurons.  (4) HK2 expression increased at all time points, and peaked at 6 to 12-fold greater than controls, respectively, at 7 days in the ipsilateral cortex and hippocampus.  GPR81 mRNA increased after 3 days in the ipsilateral cortex, whereas it decreased at day 1 in the ipsilateral hippocampus and increased after 7 days. 

Conclusions: The observed alterations in gene expression in this mouse model of TBI are novel, and correspond closely to transient increases and subsequent prolonged decreases in glucose utilization observed in TBI patients. The molecular mechanisms underlying these changes remain to be elucidated, and can inform development of new therapeutic strategies for patients with TBI.


Nothing to Disclose: JZ, MB, RA, JMS, MRB

25710 17.0000 FRI 493 A Experimental Traumatic Brain Injury in Mice Alters Cortical and Hippocampal Expression of Genes Related to Glucose Metabolism 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 3:15:00 PM FRI 476-494 7669 1:15:00 PM Neuroendocrinology (posters) Poster

Marilyn H Perrin*1, Kathy A. Lewis2, Joan M Vaughan1, Judit Erchegyi1, Charleen Miller1, Jean E F Rivier1 and Paul E Sawchenko1
1The Salk Institute for Biological Studies, La Jolla, CA, 2The Salk Institute for Biological Studies, La Jolla


CART is a peptide identified on the basis of psychotropic drug-induced upregulation of its transcript in rodent brain.  It occurs in 42- and (N-terminally extended) 48-residue forms, which have been implicated in the control of appetite/obesity, reward/addiction and stress/anxiety.  Such pathological associations have prompted interest in CART receptor(s) as potential drug targets, though no receptor has yet been identified.  Previous radioreceptor studies have used 125I-CART(61-102) to characterize binding sites in AtT-20, PC-12 and primary nucleus accumbens cells, which although of high affinity tend to be low in number, and have not yet led to the identification of a CART receptor.  To advance this effort, we have developed an analog of CART(55-102) as a radioligand, and have used it to identify binding sites on additional cells and tissues.  In order to obtain unambiguously radioiodinated ligands, we synthesized and compared one analog in which Tyr-62 was replaced by Thr, i.e., [Thr62,Nle67] CART(55-102), and another in which Tyr-58 was replaced by Thr, i.e., [Thr58,Nle67]CART (55-102). In both analogs the Met-67 was replaced by Nleu.  Following iodination using standard mild oxidation procedures and HPLC purification, the analog  [125ITyr58,Thr62,Nle67] CART(55-102) displayed significantly higher specific binding compared to the analog [Thr58,125ITyr62,Nle67]CART (55-102).  Using the preferred, Tyr58-radiolabeled, tracer (referred to hereafter as CART*), we find specific, high affinity binding to crude membranes from porcine anterior pituitary, rat hypothalamus, rat cerebellum, rat anterior pituitary, as well as from INS-1 (pancreatic islet) and N2a (neuronal) cells.  From saturation data on porcine anterior pituitary membranes the Kd for the CART* is ~0.8 nM and Bmax is 200 fmol/mg, the latter value being greater than those reported previously for AtT-20 or PC-12 cells.  Competitive displacement data for CART(55-102) binding to porcine anterior pituitary membranes revealed both low ( Ki ~200 nM) and high (Ki ~1 nM) affinity sites.  The binding of CART* is inhibited by cations (Na+, Mg++ and Ca++).  Association of CART* reaches equilibrium within 10-30 min and dissociation is nearly complete by 5 min.  We suggest that this new radioligand offers significant advantages for CART receptor studies in terms of the affinity and number of binding sites detected on diverse tissues and cells.


Nothing to Disclose: MHP, KAL, JMV, JE, CM, JEFR, PES

27026 18.0000 FRI 494 A A New Radioligand Identifies CART Binding Sites in Multiple Cells and Tissues 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Neuroendocrinology and Pituitary Friday, April 1st 3:15:00 PM FRI 476-494 7669 1:15:00 PM Neuroendocrinology (posters) Poster

Gyung Min Lee*1, Yang-Hee Park2, Sung-Jae Lee2, Chae-Hyuk Choi2, Jeong-Hoon Seo3 and June-Bum Kim2
1Konyang University Hospital, Daejeon, Korea, Republic of (South), 2Hallym University Hangang Sacred Heart Hospital, Seoul, Korea, Republic of (South), 3Hallym University Hangang Sacred Heart Hospital, Seoul


Channelopathies are diseases that develop because of defects in ion channels caused by either genetic or acquired factors (1). Familial hypokalemic periodic paralysis (HOKPP) is one of the most common inherited channelopathies and is characterized by episodic attacks of flaccid paralysis with concomitant hypokalemia (2). Patients with HOKPP experience reversible immobility throughout their lifetime and they are at increased risk for sunlight deprivation, which may lead to vitamin D deficiency. A relationship between immobility and osteoporosis is well established. However, bone changes and vitamin D status in this particular population suffering from recurrent paralysis have not been available to date. The aim of this study was to evaluate bone mineral density (BMD) and its association with age, gender, anthropometric variables, serum biochemical indices, and clinical characteristics of patients with HOKPP. Two hundred thirty-eight genetically confirmed patients with HOKPP and 238 age- and sex-matched control subjects enrolled in the Hallym Channelopathy Study, a longitudinal case-control study, were included. BMD was assessed by dual energy X-ray absorptiometry of the lumbar spine, femoral neck, and total body. Serum samples were analyzed for calcium, phosphorus, alkaline phosphatase, 25-hydroxyvitamin D [25(OH)D], and intact parathyroid hormone levels. The mean age was 29.2 (range, 6–68) years. 48.7% of patients presented with a BMD below the expected range for age in at least one of the examined sites compared with 7.1% of controls (P<0.05). 25(OH)D levels were deficient (<20 ng/mL) in 95.4% of patients compared with 67.2% of controls (P<0.01) in summer-fall season and 99.6% of patients compared with 79.4% of controls (P<0.01) in winter-spring season. A strong positive correlation was found between age at onset of the disease and BMD Z-scores at the femoral neck of patients (r=0.78, P<0.01), whereas a negative correlation was observed between disease severity (frequency and duration of paralytic attacks) and the patients’ femoral neck bone density (r=-0.81 and r=-0.63, P<0.01, respectively). 25(OH)D levels in both summer-fall and winter-spring seasons correlated positively with BMD Z-scores at lumbar spine (r=0.59 and r=0.58, P<0.01, respectively) and at femoral neck (r=0.56 and r=0.54, P<0.01, respectively). To our knowledge, this is the first report of vitamin D status and BMD in patients with familial periodic paralysis, a chronic reversible immobility. Given the high prevalence of vitamin D deficiency and low BMD observed in this study, routine measurements of vitamin D and BMD in patients with HOKPP may help to initiate therapeutic interventions to prevent these abnormal conditions. Moreover, the disproportionally high prevalence of reduced BMD warrants additional studies in other channelopathies characterized by chronic reversible attacks of impaired mobility.


Nothing to Disclose: GML, YHP, SJL, CHC, JHS, JBK

24172 1.0000 FRI 324 A Unrecognized High Prevalence of and Risk Factors for Low Bone Mineral Density in Patients with an Inherited Channelopathy 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 324-348 7679 1:15:00 PM Parathyroid Disorders and Metabolic Bone Disease (posters) Poster

Tayyab S Khan1, Hamid Syed2, J. E.M. Young3 and Aliya Aziz Khan*4
1McMaster University, HAMILTON, ON, Canada, 2McMaster University, 3McMaster University, Hamilton, Canada, 4McMaster University, Hamilton, ON, Canada


Hypoparathyroidism is an uncommon endocrine disease characterized by absent or inappropriately low levels of Parathyroid hormone (PTH), with low serum calcium and elevated phosphate levels, and presents unique therapeutic challenges. While evidence gathered over the last decade has lead to an increased understanding of this condition, data regarding disease manifestations in the Canadian population are lacking.

We established a Canadian registry of patients with hypoparathyroidism and reviewed baseline data with respect to etiology, presenting symptoms, current treatment and complications of this condition. The study was approved by the Research Ethics Board at McMaster University and all patients provided informed consent.

Most patients (53/72; 73.6%) had postsurgical hypoparathyroidism, followed by idiopathic/autoimmune disease (16/72; 22.2%) and pseudohypoparathyroidism (3/72; 4.2%).The mean age of onset was 42.8 years. Paresthesias in the upper and lower extremities were the most common presenting symptom and were experienced by 53.8% of patients. 15.4% of patients reported tetany and 13.5% had seizures. 44.2% of patients required hospitalization at the time of presentation. Current treatment options were reviewed and almost all patients were receiving calcium supplements (93.1%), 88.9% were on calcitriol, 36.1% were on hydrochlorothiazide and 5.6% were receiving parathyroid hormone. Complications were reviewed and 11 patients had brain imaging completed out of whom 9 (81.8%) had evidence of basal ganglia calcification. 13 of 32 (40.6%) patients who received an abdominal ultrasound showed evidence of nephrolithiasis or nephrocalcinosis. All patients developed nephrolithiasis or basal ganglia calcification despite a calcium phosphate product of <4.4 mmol2/L2. Out of the 17 patients for whom fracture risk was calculated using the Canadian Association of Radiologists and Osteoporosis Canada Risk Assessment Tool, 11 patients were at low risk (<10% risk of fracture over 10 years), 3 were at moderate risk (10 - 19% risk of fracture over 10 years) while 3 were at high risk of fracture (>20% risk of fracture over 10 years). All patients in the moderate or high risk groups had traditional risk factors for osteoporosis.

Our findings provide interesting insights into the etiology, symptomatology, treatment strategies and complications of hypoparathyroidism in a Canadian population.


Disclosure: AAK: Clinical Researcher, NPS, Clinical Researcher, Amgen, Clinical Researcher, Merck & Co.. Nothing to Disclose: TSK, HS, JEMY

25242 2.0000 FRI 325 A An Overview of the Etiology, Clinical Manifestations, Management Strategies and Complications of Hypoparathyroidism from the Canadian National Hypoparathyroidism Registry 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 324-348 7679 1:15:00 PM Parathyroid Disorders and Metabolic Bone Disease (posters) Poster

Kamal Bhusal* and Harsha Karanchi
LSU Health Shreveport, Shreveport, LA


Background: Chronic lithium use is associated with various endocrinopathies including hyperparathyroidism (10 -20 %), nephrogenic diabetes insipidus (20-40%), goiter, hypothyroidism and hyperthyroidism. The co-occurrence of multiple lithium related endocrinopathies is rare.

Clinical Case: A 63-year woman with mental retardation and bipolar disorder on lithium therapy for at least three years developed multinodular goiter and hyperparathyroidism. She was initially seen by ENT and underwent left hemithyroidectomy and removal of left superior parathyroid gland which on pathology revealed parathyroid adenoma and incidental micro papillary thyroid cancer.  Hypercalcemia and hyperparathyroidism did not resolve after surgery. She underwent completion thyroidectomy with right superior parathyroidectomy showing parathyroid adenoma.  Even after removal of 2 parathyroid glands, hyperparathyroidism persisted and she was referred to our endocrine for further evaluation and management. Her albumin corrected calcium was high at 10.9 mg/dL (nl. 8.5-10.1 mg/dL) with PTH level of 74.3 pg/mL (nl. 12.4-76.8), normal phosphorus of 3.9 mg/dL (nl.2.5-4.9) Vitamin D insufficiency with 25-OH Vitamin D of 26.14 ng/mL (nl. > 30 ng/mL), high 1,25 Vitamin D level of 130.4 pg/mL (nl. 10-75 pg/mL)  and hypocalciuria on 24-hour urine calcium of 23 mg/24 hours. She reported polyuria and polydipsia and on work up showed hypotonic polyuria and hypernatremia suggestive of diabetes insipidus (DI). With coordination of psychiatry team, lithium was discontinued. But even 6 months after discontinuation of lithium, both hyperparathyroidism and DI have not resolved and follow up is ongoing

Conclusion: Lithium use can cause multiple endocrinopathies which can persist even after discontinuation of lithium. In our patient, hypercalcemia, hyperparathyroidism and DI persisted. Lithium associated hyperparathyroidism has higher prevalence of multiglandular disease than sporadic hyperparathyroidism and surgery without bilateral neck exploration and 3 ½ gland parathyroidectomy may not be curative. It is important to be vigilant and monitor for the co-existence of hypercalcemia and nephrogenic DI related to lithium use as dehydration can exacerbate hypercalemia.


Nothing to Disclose: KB, HK

26259 3.0000 FRI 326 A Persistent Hyperparathyroidism s/p Two Gland Parathyroid Adenomectomy, Goiter and Nephrogenic DI Associated with Lithium Use 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 324-348 7679 1:15:00 PM Parathyroid Disorders and Metabolic Bone Disease (posters) Poster

Verena Schwetz*1, Martin Kern1, Andrea Bradatsch2, Christian Trummer1, Gerhard Wolf2, Thomas R Pieber3, Jutta Claudia Piswanger-Soelkner1, Harald Dobnig4 and Karin Amrein1
1Medical University of Graz, Graz, Austria, 2Medical University of Graz, 3Medical University of Graz, Austria, 4Thyroid Endocrinology Osteoporosis Institute Dobnig, Graz, Austria



Chronic hypoparathyroidism is a hormone disorder that is typically caused by removal or damage to the parathyroid glands during or after neck surgery. Other less frequent causes include genetic and autoimmune disorders. Hypoparathyroidism substantially impacts quality of life for many patients because it causes a multitude of symptoms and requires chronic medication. However, the degree of decreased quality of life of patients with hypoparathyroidism is often underestimated. Current standard therapy is only symptomatic using primarily calcium and native/active vitamin D in greatly varying individual doses. There are to date no guidelines and hypoparathyroidism remains one of the last endocrinopathies not being treated with the missing hormone. In the future the parenteral application of parathyroid hormone may play a role.


In this retrospective study, 120 patients with hypoparathyroidism were identified at the Division of Endocrinology and Metabolism at the Medical University of Graz between 2004 and 2015, using the routine patient data management system. Patients were included if they had parathyroid hormone levels in the low or below the normal range (<25 pg/ml) in addition to a relevant diagnosis.


The cohort included mostly women (n=94, 78 %). The mean age at diagnosis was 55 years. Mean total serum calcium was 2.09 mmol/l +/- 0.32. 46 % of the patients had hypocalcemic and 2.5 % hypercalcemic values. Paraesthesia and tetany were significantly more frequent in patients with low ionized calcium (p<0.05). This was not the case for low total serum calcium levels (p=0.53). Overall, 66 % complained about tetany at least once and therapy was significantly more often modified in these patients (p<0.01). 88 % of the patients had postoperative hypoparathyroidism. Although almost half of the patients were diagnosed within the first year after surgery, median time from surgery to diagnosis was 5.5 years with a maximum of more than 60 years, making a second hit-theory (e.g. radioiodine therapy) at least in some patients likely.


The high rates of tetany and paraesthesia suggest that patients with persistent hypoparathyroidism often remain symptomatic despite current standard treatment. Individualized therapy also based on clinical symptoms with sufficient doses of calcium, cholecalciferol and calcitriol may lead to substantial improvement. We suggest that in many cases of postoperative etiology, a second hit such as radioiodine therapy or atherosclerosis leads to overt hypoparathyroidism.


Nothing to Disclose: VS, MK, AB, CT, GW, TRP, JCP, HD, KA

25323 4.0000 FRI 327 A Hypoparathyroidism - Descriptive Results of a Large Retrospective Austrian Cohort 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 324-348 7679 1:15:00 PM Parathyroid Disorders and Metabolic Bone Disease (posters) Poster

Angelica Maria Silva1, Lily J Kwatampora*2, Ioannis Christakis3, Carla Warneke4, Michelle D Williams4, Callisia Clarke1, Elizabeth G Grubbs3, Jeffrey E Lee5, Nancy D Perrier3 and Naifa L Busaidy3
1University of Texas MD Anderson Cancer Center, Houston, 2Lutheran Health Physicians, Fort Wayne, IN, 3The University of Texas MD Anderson Cancer Center, Houston, TX, 4University of Texas MD Anderson Cancer Center, Houston, TX, 5University of Texas, MD Anderson Cancer Center, Houston TX, TX



Parathyroid carcinoma (PC) is a rare malignancy and surgery remains the mainstay of treatment. There are no universally recognized prognostic factors that consistently predict recurrence and survival. Due to the rarity of disease and limited therapeutic options, examining these factors are essential in informed treatment decisions. We report the largest single Institution experience over the last 3 decades and aim to describe clinicopathologic features associated with recurrence and poor survival.


We performed a retrospective chart review of all histopathologically diagnosed PC patients operated/treated in our Institution (1980-2015), excluding those with tumors of uncertain malignancy and atypical neoplasms. Persistent cases were those who were operated and developed disease or didn't normalize the calcium within 6 months postoperatively. Demographics, clinical characteristics, histopathologic features and survival rates were analyzed. Overall survival (OS) was calculated from PC diagnosis until death or until date of last known vital status. Disease-free survival (DFS) was calculated from the date of the surgery first rendering the patient disease free for ≥ 6 months or until last clinical follow-up (FU) for patients still in their first disease-free interval at end of FU. P-values < 0.05 were considered significant.


PC was identified in 52 patients (mean age 52.7 years, SD 15.1). Males represented 52% (n=27) and females 48% (n=25). The most common symptom was fatigue (n=18), followed by abdominal symptoms (n=11) and memory loss (n=8). Median PTH was 507 pg/ml (range 67–7200), and median highest calcium level was 13.2 mg/dl (range 9.7–20.5). Mean tumor size was 2.71 cm (SD 0.95) and mean tumor weight was 6.19 g (SD 4.26). Vascular invasion present in 16 patients (31%) had no effect on DFS or OS. Median OS was 13.77 years (95% CI 6.76, 24.25). Five-year OS was 80.71% (95% CI 63.46%-90.40%), and 10-year OS was 54.88% (95% CI 34.00%, 71.63%). In univariate analysis, OS was inversely associated with age at diagnosis (HR 1.06, 95% CI 1.02, 1.10, p=0.0022). Patients with a disease-free interval < 6 months had poorer OS compared to OS of patients rendered disease free at first surgery (p=0.0077). Median DFS was 6.02 years (95% CI 1.91, not attained). Five-year DFS was 59.61% (95%CI 40.76%-74.22%). Type of initial surgery (parathyroidectomy alone or en bloc resection) was not associated with OS or DFS.


PCs demonstrated a worse prognosis in patients who were older at diagnosis or had persistent disease following initial surgery (residual disease). Vascular invasion did not have a significant effect on recurrence or survival. Complete, but not necessarily radical, surgery at the time of the first operation for PC could help prevent disease persistence and improve OS. A larger cohort is needed to further identify prognostic clinicopathologic characteristics.


Nothing to Disclose: AMS, LJK, IC, CW, MDW, CC, EGG, JEL, NDP, NLB

24868 5.0000 FRI 329 A Prognostic Factors of Overall Recurrence and Survival in Parathyroid Carcinoma; The Largest Single Institution Experience over the Last 3 Decades in USA 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 324-348 7679 1:15:00 PM Parathyroid Disorders and Metabolic Bone Disease (posters) Poster

Regina Belokovskaya*1, Francisco Perez Mata2 and Oksana Davydov1
1Mt. Sinai St. Luke's Roosevelt Hospital, New York, NY, 2Mount Sinai St. Luke's and Mount Sinai Roosevelt Hospitals, New York, NY


Introduction: Sarcoidosis is a chronic multisystem granulomatous disease of unknown etiology, which is characterized by the presence of noncaseating granulomas in involved organs. Of the patients diagnosed with sarcoidosis, 35 to 50 percent develop renal complications. Patient described here, was found to have sarcoidosis when he presented with acute renal failure, hyperkalemia, and mild hypercalcemia.

Clinical Case: A 55-year-old male with past medical history of right obstructing kidney stone and herpes zoster with post-herpetic neuralgia was referred to the emergency room for elevated creatinine. Few weeks prior to this admission, the patient  received calamine lotion and Tylenol treatment during his trip to Dominican Republic. He complained of unintentional weight loss over the last four months and a burning pain on the left chest wall at the site of the herpes zoster infection. On exam, he was hemodynamically stable with well crusted vesicular lesions in left-sided T2-T3 distribution on chest, back, and axilla. His laboratory values were significant for Creatinine of 7.59 mg/dL (0.66-1.25), BUN of 57 mg/dL (8-24), Sodium of 135 mmol/L (136-146), Potassium of 5.3 mmol/L (3.5-5.1), Calcium of 13.5 mg/dL (8.4-10.3), Phosphorus of 7.4 mg/dL (2.5-4.5), intact PTH of 3.72 pg/mL (11-67), Vitamin D 25-OH of 23.8 ng/dL (30-95), Hemoglobin A1C of 6.9 % (4.2-5.9). Hemoglobin and hematocrit were within normal limits. The complements came back within normal limits with negative ANA. HIV and Hepatitis C were negative, with reactive Hepatitis A Ab, Hepatitis B core and surface Ab.  Urinalysis showed moderate blood, glucose of 500, protein of 100, and a presence of moderate Calcium Oxalate crystals. Chest X-ray was negative for any acute pulmonary process. Ultrasound of the bilateral kidneys did not reveal any stones or hydronephrosis. Skeletal survey did not identify any lytic or blastic osseous lesions. Despite Normal Saline infusion at 150 cc/hr, Calcium remained elevated. Angiotensin-converting enzyme came back elevated at 82 (9-67 U/L). CT chest without IV contrast demonstrated airspace opacities in the right middle and lower lobes along with interlobular septal thickening. IR guided kidney biopsy showed granulomatous interstitial nephritis with diffuse interstitial inflammation including eosinophils - compatible with sarcoidosis. Initially, patient was started on pulse steroids, Solumedrol 40 mg every 6 hours IV and then PO Prednisone 1mg/kg. At the completion of his hospital stay, patient’s Creatinine decreased to 2.89, Calcium normalized at 9.6, and Potassium at 4.9. He was discharged on Prednisone 60 mg PO daily with close renal, endocrine and pulmonary follow-up.

Conclusion: Non-PTH mediated hypercalcemia, hyperkalemia, and acute renal failure in an asymptomatic patient should prompt a work-up for sarcoidosis.


Nothing to Disclose: RB, FP, OD

27518 6.0000 FRI 330 A Sarcoidosis Presenting with Acute Renal Failure, Hypercalcemia and Hyperkalemia 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 324-348 7679 1:15:00 PM Parathyroid Disorders and Metabolic Bone Disease (posters) Poster

Anand Vaidya*1, Gary Curhan2, Julie Paik3, Molin Wang4 and Eric Taylor2
1Brigham and Women’s Hospital/Harvard Medical School, Boston, MA, 2Brigham and Women's Hospital, Harvard Medical School, 3Brigham and Women's Hospital, Harvard Medical School, Brookline, MA, 4Harvard School of Public Health/Brigham and Women's Hospital


Importance:  P-HPTH is a relatively common disorder that predominantly affects women. Our understanding of modifiable risk factors for developing primary hyperparathyroidism (P-HPTH) is limited. 

Background: Prior studies have shown that physical activity (PA) can lower parathyroid hormone (PTH) levels, but there are no prospective studies evaluating whether PA influences the risk for developing P-HPTH.

Objective: To evaluate the hypothesis that lower PA is a risk factor for developing P-HPTH.

Design/Setting/Participants: A prospective study of 69,621 female participants in the Nurses’ Health Study I, without baseline P-HPTH, followed for 22 years from 1986 to 2008.

Exposures: PA and other dietary and demographic exposures were quantified via detailed, and validated, biennial questionnaires.

Main Outcome Measures: Incident P-HPTH was confirmed by individual medical record review after initial assessment by questionnaire. Adjusted Cox proportional hazards models were used to evaluate whether PA was an independent risk factor for developing P-HPTH. We also evaluated the risk of developing P-HPTH when combining low PA (<16 MET hours/week) with previously identified independent risk factors for developing P-HPTH: low calcium intake (<800 mg/d) and diagnosis of hypertension (Y/N).  In a subset of participants who had biochemical measurements taken for research purposes (n=625), we evaluated the independent relation between PA and PTH levels.

Results: We confirmed 302 incident cases of P-HPTH during 1,474,993 person-years of follow-up.  Participants in the highest quintile of PA had a 50% lower risk of developing P-HPTH: the age-adjusted relative-risks and 95% confidence intervals for incident P-HPTH by lowest to highest quintile of PA were Q1=1.0 (reference); Q2=0.83 (0.60, 1.15); Q3=0.84 (0.61, 1.15); Q4=0.50 (0.34, 0.74); Q5=0.50 (0.35, 0.73); P-trend<0.001. Extensive multivariable adjustments did not materially change these findings. When compared to participants with no established risk factors for P-HPTH (high PA, high calcium intake, and no hypertension), the adjusted relative risk for incident P-HPTH in those with 1, 2, and all 3 risk factors was 1.56 (1.04, 2.34), 2.32 (1.54, 3.50), and 4.16 (2.49, 6.95).  PA was inversely correlated with serum PTH (ρ= -0.09, P=0.03) and the mean adjusted serum PTH declined from 39.1 to 35.3 pg/mL from lowest to highest quintile of PA (P-trend<0.01).

Conclusion: Low physical activity may be a modifiable risk factor for developing P-HPTH in women. The combination of low physical activity, low calcium intake, and hypertension may be characteristics that predispose women to a greater than 4-fold higher risk of developing P-HPTH. These findings may influence future studies focused on the prevention, or mitigation, of P-HPTH.

Disclosures: None


Nothing to Disclose: AV, GC, JP, MW, ET

24086 7.0000 FRI 331 A Physical Activity and the Risk of Developing Primary Hyperparathyroidism 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 324-348 7679 1:15:00 PM Parathyroid Disorders and Metabolic Bone Disease (posters) Poster

Namita G. Hattangady*, Tremika Le-Shan Wilson, Barbra S. Miller, Antonio M Lerario, Thomas J Giordano, Palak Choksi and Tobias Else
University of Michigan, Ann Arbor, MI


Background: Primary hyperparathyroidism (pHPT) is commonly caused by adenomas, less frequently by multiglandular parathyroid hyperplasia and rarely by parathyroid cancer. pHPT is mostly sporadic but occurs as part of familial syndromes such or multiple endocrine neoplasia type 1 (MEN1) or Hyperparathyroidism-Jaw Tumor (HPT-JT) syndrome. HPT-JT is caused by inactivating mutations in the tumor suppressor gene CDC73 (or HPRT2, which codes for the protein parafibromin). Herein we investigate an intronic germline mutation in CDC73 in a patient with recurrent pHPT despite multiple surgeries.

Clinical case: The index case is a 52 year old male with a long standing history of kidney stones who was diagnosed with pHPT at age 35. The initial surgery removed two hyperplastic parathyroid glands resulting in normalization of calcium levels. Seven years later, the patient had recurrent pHPT and underwent removal of two adenomatous glands and autotransplantation to the forearm. Since then he had recurrence of pHPT and unsuccessful surgery to reduce parathyroid tissue in the forearm. The probands’s family history was negative for pHPT, jaw tumors or kidney lesions. The proband’s mother, however, did undergo a hysterectomy, possibly due to uterine fibroids. Initial genetic testing did not reveal a mutation in MENIN, but revealed the presence of a germline intronic variant of uncertain significance (VUS) in CDC73c.238-8G>A (IVS2-8G>A).

Methods: In order to gather in vitro evidence for pathogenicity of the mutation, we analyzed loss of heterozygosity and gene splicing as the mutation was predicted to activate a cryptic splice site, resulting in early translation termination. Genomic DNA from formalin fixed paraffin embedded parathyroid tumor tissues showed loss of heterozygosity, while it was preserved in control thyroid tissue. Immunostaining for parafibromin confirmed protein expression in thyroid tissue and loss of expression in the parathyroid adenoma tissues. With regards to an effect on splicing, both predicted transcripts were found by RT-PCR in mRNA from peripheral blood. In vitro characterization of the splice variant was confirmed by cloning the genetic fragment containing the wildtype and mutant intronic region and its successive exon into the pDUP4-1 minigene vector and expressing it in human embryonic kidney (HEK293 cells) by transfection. Cells expressing the pDUP4-1/Mutant CDC73 displayed a splice variant of CDC73 which was 6 base pairs longer than the wild type CDC73 mRNA.

Conclusion: In summary, we show that this novel intronic CDC73 mutation, c.238-8G>A, activates a cryptic splice site and that the wild type allele is lost in this patient’s parathyroid adenomas. Together, these findings provide in vitro evidence to reclassify this mutation as a pathogenic variant confirming our patient’s diagnosis of CDC73-related disorder and an increased risk for parathyroid cancer development.


Nothing to Disclose: NGH, TLSW, BSM, AML, TJG, PC, TE

25825 8.0000 FRI 332 A Recurrent Hyperparathyroidism Due to a Novel CDC73 Splice Mutation 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 324-348 7679 1:15:00 PM Parathyroid Disorders and Metabolic Bone Disease (posters) Poster

Leonardo Bandeira*1, Diane Cozadd1, Mariana Bucovsky1, Donald J McMahon1, James A Lee2, Shonni J. Silverberg1 and Marcella D. Walker1
1Columbia University, College of Physicians and Surgeons, New York, NY, 2Columbia University Medical Center, New York, NY


CONTEXT: Currently up to 80% of patients with primary hyperparathyroidism (PHPT) are asymptomatic in the United States (US) and other parts of the world where routine biochemical screening of calcium occurs. While symptomatic nephrolithiasis occurs in approximately 17% of PHPT patients in the US, the incidence of occult stones is unknown. However, the most recent (2014) guidelines for the evaluation and management of asymptomatic PHPT suggest renal imaging to detect occult nephrolithiasis and referral for parathyroidectomy (PTX) when occult stone disease is present.

OBJECTIVE: We sought to determine the prevalence of occult renal stones in patients with PHPT and determine clinical risk factors associated with this finding.

METHODS / RESULTS: PHPT patients at our institution are now routinely sent for renal imaging (abdominal/renal ultrasound, x-ray, CT or MRI) as part of their clinical evaluation. We are prospectively collecting data on patients without a clinical history of nephrolithiasis, including demographic data, medical history, biochemistries (serum 25-hydroxyvitamin D (25OHD), 1,25-dihydroxyvitamin D, phosphorus, estimated glomerular filtration rate (eGFR), calcium, albumin, parathyroid hormone and urinary calcium and creatinine) and radiology results. To date, 38 PHPT patients have been enrolled. Participants (89.5% female, 65.8% white, mean age 62.6±11.5 yrs) had biochemically mild PHPT (mean calcium 10.6±0.4mg/dl, PTH 84.9±44pg/ml, 25OHD 29.4±12.7ng/ml). The majority of patients (57.9%) met at least one criterion for PTX based on 2008 guidelines. Occult kidney stones were identified by imaging in 5 patients (13.2%, 95% CI: 5-28%). Those with vs. without nephrolithiasis had higher mean daily vitamin D supplement intake (2040±669 vs. 870±938 IU/day, p=0.01), higher 1,25-dihydroxyvitamin D levels (100.6±26.2 vs. 57.2±18pg/ml, p=0.05) and greater urine calcium excretion (297±119 vs. 174±82 mg/g creatinine, p=0.01). There were no between-group differences in age (68±8 vs. 62±12 yrs, p=0.2), weight (165±31 vs. 165±51 lbs, p=0.9), race (80% vs. 63.6% white, p=0.4), serum calcium (10.6±0.2 vs. 10.6±0.4 mg/dl, p=0.7), PTH (74±31 vs. 87±46pg/ml, p=0.6), 25OHD level (36.7±10.9 vs. 28.6±12.8ng/ml, p=0.2), serum phosphate (3.5±0.5 vs. 3.2±0.4mg/dl, p=0.2), renal function (eGFR: 78±8 vs. 81±22ml/min, p=0.8), BMD by DXA at any site or rates of osteoporosis (40% vs. 37.5%, p=1).

CONCLUSION: Occult nephrolithiasis is not uncommon among patients with asymptomatic PHPT. Nephrolithiasis was associated with greater vitamin D supplement intake, higher serum levels of 1,25-dihydroxyvitamin D and higher urinary calcium excretion. These preliminary data support the indication for renal imaging in asymptomatic PHPT and suggest that higher vitamin D intake may predispose to higher activated vitamin D levels, urinary calcium excretion and occult nephrolithiasis in PHPT.


Nothing to Disclose: LB, DC, MB, DJM, JAL, SJS, MDW

24218 9.0000 FRI 333 A Occult Nephrolithiasis in Primary Hyperparathyroidism 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 324-348 7679 1:15:00 PM Parathyroid Disorders and Metabolic Bone Disease (posters) Poster

Oliver Tschopp*, Kathrin Roost and Christoph Schmid
University Hospital Zurich, Switzerland


Introduction: Brown tumors (BT) are a rare manifestation of long standing primary hyperparathyroidism (pHPT), and the severity of the disease is an indication for parathyroid surgery. Following removal of the dominant PTH source, "hungry bone syndrome" (HBS) with prolonged hypocalcemia is nowadays rare in vitamin D-replete patients with pHPT, but appears to be more common in the subgroup with BT. This unusual presentation has led to a large number of individual case reports, but original series of several patients are scarce.

Methods: We describe 8 (6 female) patients with pHPT presenting with BT and compared them to a control group (n=16, 12 female) with pHPT without BT (presenting with previously detected hypercalcemia), matched not only for gender but also for place and year of surgery. Skeletal involvement at presentation was readily detected by bone biopsy or imaging (scintigraphy, MRI) and in 5 of 8 patients also by 18F-PET/CT. Patients were treated by parathyroidectomy, resulting in correction of hypercalcemia in all of them.

Results: At evaluation, patients with pHPT and BT were younger (44.6±4.9 vs. 55.4±3.3 years; mean ± SEM) and presented with significantly (p<0.05 by two-tailed t-test) lower BMI (20.2±1.4 vs. 26.8±1.6 kg/m2), higher calcium (3.4±0.2 vs. 2.9±0.1 mmol/l), PTH (878±186 vs. 154±19 ng/l) and alkaline phosphatase (AP) (334±109 vs. 92±7 U/l) serum levels; there was no significant difference in serum creatinine between the two groups (95±12 vs. 86±6 µmol/l). Adenomas were readily detected by imaging and/or by the surgeon and were significantly larger in patients with BT (weighing 5.4±1.6 vs. 1.2±0.4 grams). Following surgery, 5 of 8 patients with BT (none of the controls) developed HBS requiring prolonged iv calcium treatment, resulting in a significantly longer hospital stay (21.0±5.7 vs. 3.8±0.8 days). Hypocalcemia was accompanied by hypophosphatemia, low urinary calcium as well as persistently high AP. Skeletal recovery was reflected by a decrease in AP activity towards normal, by reappearance of normal amounts of calcium in fasting spot urine samples and the feasibility of an oral treatment preventing symptomatic hypocalcemia (sufficiently effective to permit dismission from the hospital). Permanent treatment with calcitriol was necessary in 3 of 24 patients (1 with BT)

Conclusions: We conclude that presentation of pHPT with BT constitutes an increased risk for development of a postoperative HBS with prolonged need for iv calcium treatment to correct and prevent severe hypocalcemia. Predictors of such a complicated postoperative course include not only high PTH but also high calcium and AP serum levels at presentation. In contrast to previous reports (with less strict definition of HBS), our patients presenting with BT were younger than the controls (presenting with hypercalcemia).


Nothing to Disclose: OT, KR, CS

24502 10.0000 FRI 334 A Hungry Bone Syndrome after Surgery for Primary Hyperparathyroidism in Patients with Brown Tumors - a Case Series 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 324-348 7679 1:15:00 PM Parathyroid Disorders and Metabolic Bone Disease (posters) Poster

Thomas Ian Hadwen*1, Melissa Clarke1, Luke Conway2, Anna-Gerardina Mclean3 and Ashim Kumar Sinha4
1Royal Brisbane and Women's Hospital, Brisbane, 2Cairns Hospital, Cairns. QLD, Australia, 3Cairns Hospital, Cairns, Australia, 4Cairns Hospital. James Cook University, Cairns QLD, Australia



Primary hyperparathyroidism is a common endocrinological disease with an evolving clinical presentation.  Symptomatic disease was common but currently the majority of diagnoses are made in asymptomatic patients.  Current guidelines are for surgical treatment if patients are symptomatic or if asymptomatic and meet set criteria, including a BMD T score of less than -2.5.  There is conflicting data in the literature on the correlation between BMD and other variables associated with PHPT such as PTH, corrected calcium (CCa), eGFR, phosphate, 25-vitamin D (25-VitD) and 1,25-vitamin D (1,25VitD).  Previous cross-sectional studies had shown a direct relationship between 25-VitD and BMD at all sites whereas others had shown significant results only at the hip but not at the lumbar or distal radius (1,2).  In contrast to 25-VitD, 1,25VitD has been shown to have an inverse correlation with BMD, as has PTH (3).


We aimed to analyse the data of patients with PHPT looking for correlations between BMD, PTH, CCa, eGFR, phosphate, 25-VitD and 1,25VitD.


We performed a prospective case series of all patients referred to our regional hospital with hypercalcaemia and subsequently diagnosed with PHPT between 2013 and 2015.


65 patients were diagnosed with PHPT.  There were 20 males and 45 females with an average age of 62.  47 patients had a BMD performed at the time of diagnosis.  The average lumbar BMD T-score was -1.17, the average femoral BMD T-score was -1.6 and this difference was statistically significant.  13% had a lumbar T-score less than -2.5 and 23% had a femoral T-score less than -2.5.  The average eGFR was 74ml/min.  The average 25-VitD was 24.4ng/ml with 31% of patients being vitamin D deficient (<20ng/ml) but no patients having severe vitamin D deficiency (<10ng/ml).  The average 1,25VitD was 1.15 times the upper limit of normal.  Analysing the data showed that there was no statistically significant correlation between either femoral or lumbar BMD and PTH, CCa, eGFR, age, phosphate, 25-VitD or 1,25VitD.  In addition the 25-VitD level did not correlate with eGFR or age but was positively correlated with phosphate (r=0.30, p=0.04).  The 1,25VitD level was positively correlated with eGFR (r=0.46, p=0.02).


In our study BMD at both the lumbar region and femoral neck did not correlate with any of the previously associated markers for PHPT disease severity such as 25-VitD, CCa and PTH.  Vitamin D deficiency had previously been associated with a lower BMD but we did not find this.  The difference in findings could be due to the lack of severe vitamin D deficiency in our group.  Previous studies had found that the 1,25VitD level was negatively correlated with BMD despite the opposite being true for 25-VitD.  We did not find this.  In conclusion, in our prospective case series of patients with PHPT, BMD was not significantly affected by either PTH, 25-VitD or 1,25VitD.


Nothing to Disclose: TIH, MC, LC, AGM, AKS

25217 11.0000 FRI 335 A Bone Mineral Density in Primary Hyperparathyroidism: The Effects of PTH, 25-Vitamin D and 1,25-Vitamin D 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 324-348 7679 1:15:00 PM Parathyroid Disorders and Metabolic Bone Disease (posters) Poster

Yulong Li*, James Welch and William F Simonds



Calcium/creatinine (Ca/Cr) renal clearance ratio (CCRCR) test has been used to separate familial hypocalciuric hypercalcemia (FHH) from primary hyperparathyroidism (PHPT), because the former is a benign condition and rarely requires intervention.  Current guidelines have set the CCRCR cutoff value of <0.01 for diagnosis of FHH and >0.02 for PHPT. Few studies have examined the discriminative power of CCRCR in separation of FHH from PHPT, and the cut-off value of <0.0115 was reported in a Danish study to be optimal for diagnosis of FHH with a specificity of 0.88 and a sensitivity of 0.80.  No study has yet evaluated if CCRCR can distinguish FHH from HPT in the context of Multiple Endocrine Neoplasia Type 1 (MEN1) or hyperparathyroidism-jaw tumor syndrome (HPT-JT). This study was aimed at assessing the discriminatory power of CCRCR test in patients with hypercalcemia.


Among the patients enrolled in two hypercalcemia- and PHPT-related studies at our institute, we identified those who had the diagnoses of FHH, MEN1 or HPT-JT with the confirmative genetic test results as well as those with sporadic PHPT. The paired results of plasma Ca/Cr and 24h urine Ca/Cr excretion of these patients were extracted, and CCRCR was calculated as (24h U-Ca/P-Ca, total)/(24h U-Cr/P-Cr).


There were a total of 17 FHH, 62 PHPT, 8 HPT-JT, and 61 MEN1 patients with available CCRCR results from November 1982 to May 2015.  The diagnoses of FHH, MEN1 and HPT-JT were confirmed by detection of mutations in CASR, MEN1 and CDC73/HRPT2 genes, respectively. A total of 29 measurements of CCRCR were made in the FHH patients with a mean ± SD of 0.01198 ± 0.00541, 102 in PHPT (0.02331 ± 0.01347), 16 in HPT-JT (0.03238 ± 0.01758), and 136 in MEN1 (0.01832 ± 0.01195). Using the cutoff value of <0.01, the sensitivity of CCRCR test to detect FHH in all patients was 0.41, with a specificity of 0.87, positive predictive value (PPV) of 0.27, and negative predictive value (NPV) of 0.93. When using the cutoff value of <0.0115, the sensitivity improved to 0.52, with the specificity, PPV and NPV of 0.82, 0.24 and 0.94, respectively.  The optimal CCRCR cutoff value to distinguish FHH from sporadic PHPT was <0.015, which yielded a sensitivity of 0.76 and a specificity of 0.75.  The optimal cutoff value to distinguish FHH from HPT-JT was <0.0180, with a sensitivity of 0.86 and a specificity of 0.81. Due to a significant overlap of CCRCR values in FHH and MEN1, no cutoff value could be identified to distinguish between these two diseases with both sensitivity and specificity above 0.7.


Our results showed that CCRCR test has limited discriminative power to separate FHH from sporadic PHPT, or HPT in the context of HPT-JT, with an optimal cutoff point between 0.0115 and 0.0180. Although the CCRCR test cannot be used to distinguish FHH from MEN1, the high penetrance of non-parathyroid neoplasias in the latter usually makes this distinction readily apparent.


Nothing to Disclose: YL, JW, WFS

25804 12.0000 FRI 337 A The Diagnostic Value of Calcium/Creatinine Renal Clearance Ratio in Hypercalcemic Disorders: The Experience at a Single Institution 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 324-348 7679 1:15:00 PM Parathyroid Disorders and Metabolic Bone Disease (posters) Poster

Yousuf Khan* and David S Schade
University of New Mexico School of Medicine, Albuquerque, NM



Parathyroid cancers (PCs) are very rare and account for 0.005% of the total cancer cases reported to the National Cancer Data Base (NCDB). Preoperative diagnosis of PC is often difficult with up to 86% of cases not being initially appreciated even by experienced parathyroid surgeons. We present a case of primary hyperparathyroidism (HPT) which was suspected to be caused by a parathyroid adenoma but turned out to be parathyroid cancer.

Case details:

A 64 y/o female was diagnosed with primary HPT in 2004. She was a poor surgical candidate because of her multiple cardiopulmonary comorbidities. She had no family history of MEN-1 or hypercalcemia. Her neck ultrasound suggested a left superior parathyroid nodule. The patient was medically managed at this time. Because of a recent worsening of calcium levels and progressive osteoporosis, the patient underwent a FNA biopsy of the suspicious left superior parathyroid nodule. The cytology showed parathyroid hyperplasia with no malignancy. With concern for rising calcium levels, a decision was made for a neck exploration. An apparently large 2.5 cm right superior parathyroid nodule was identified during surgery. A right superior parathyroidectomy was performed. Intraoperative frozen section biopsy was consistent with hypercellular parathyroid tissue. However, the final surgical pathology showed parathyroid carcinoma with capsular and lymphovascular invasion. Tumor cells showed minimal mitotic activity and a low Ki-67 labeling index. The postoperative whole body PET-CT scan was negative for metastasis. A watchful waiting approach was adopted.


Parathyroid cancer occurs in less than 1% of patients with primary HPT. The etiology of PC is unclear. There is an increased association with MEN-1, MEN-2A, hyperparathyroidism-jaw tumor syndrome and familial isolated primary HPT. Recent evidence suggests the role of HRPT2 gene mutations in the pathogenesis of hyperparathyroidism-jaw tumor syndrome and sporadic parathyroid cancer. A diagnostic FNA biopsy is not indicated in a suspected case of PC because of risk of tumor seeding. PCs are indolent cancers with low malignant potential. In the NCDB study, the overall relative survival rate was 85.5% at 5 years and 49.1% at 10 years. Mortality results from complications of HPT rather than from metastasis. Following initial surgery, recurrence rates range from 33 to 78%. The success of surgery depends on preoperative suspicion and the surgeon’s experience.


A parathyroid cancer should be suspected clinically when: 1) serum calcium is greater than 14 mg/dl. 2) serum PTH is more than twice the upper limit of normal. 3) hypercalcemia is associated with a palpable cervical mass. 4) hypercalcemia is associated with unilateral vocal cord paralysis or 5) there is concomitant renal and skeletal disease with a high PTH levels.


Nothing to Disclose: YK, DSS

26143 13.0000 FRI 338 A Unsuspected Parathyroid Cancer: A Case Report 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 324-348 7679 1:15:00 PM Parathyroid Disorders and Metabolic Bone Disease (posters) Poster

Adam Stangierski, Kosma Wolinski and Marek Ruchala*
Poznan University of Medical Sciences, Poznan, Poland


Introduction: Shear wave elastography (SWE) is a newly introduced method of the estimation of tissue stiffness. It has already been proven to be a valuable tool in the evaluation of thyroid lesions, thyroid autoimmune disorders and thyroid inflammations. The aim of our study was to check if SWE can complement the diagnostics of primary hyperparathyroidism.

Materials and methods: Patients referred to the surgery due to diagnosed primary hyperparathyroidism were included. In all cases presence of parathyroid adenoma (PA) was confirmed by histopathology. Control group consisted patients with benign thyroid nodules referred for surgery. Benign characters of nodules was confirmed by histopathology in all cases. Maximal elasticity (Emax) of each lesion was recorded.

Results: We have included 66 patients with PA and 98 patients possessing 322 benign thyroid nodules as a control group. Mean Emaxvalue for parathyroid adenomas was 13.4 kPa with standard deviation (SD) 17.9 kPa; median was 8.0 kPa. For benign thyroid lesions mean was equal to 57.3 with SD=60.6, median – 36.2 kPa. The difference was statistically significant (p<0.05).

Conclusions: Basing upon our results PAs were significantly more elastic than thyroid lesions. We have revealed, that median stiffness of PAs was over three times lower than of thyroid lesions. This specific finding may be considered as a new marker of PAs, complementing previously described sonographic features of PAs, like hypoechogenicity or localization near the rear wall of the thyroid. Therefore, such lesions should indicate further biochemical evaluations of potential hyperparathyroidism.


Nothing to Disclose: AS, KW, MR

26237 14.0000 FRI 339 A Shear-Wave Elastography in Diagnostics of Primary Hyperparathyroidism, a Novel Application of the Method 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 324-348 7679 1:15:00 PM Parathyroid Disorders and Metabolic Bone Disease (posters) Poster

Betiana Mabel Perez*1, Maria Pia Podesta1, Rodrigo Serrano Salinas1, Giovana Rosario Córdoba Figueroa1, Magdalena Pavlove1, Silvia Viviana Karlsbrum1 and Helena H Salerni2
1Durand Hospital, Buenos Aires, Argentina, 2CICEMO, Buenos Aires, Argentina


Vitamin D deficiency (VDD) is common in patients with primary hyperparathyroidism (PHPT), and this could affect the clinical expression of the disease. Fear of potential hypercalcemia makes it difficult to convince medical community to supplement vitamin D (VD) to hypercalcemic patients. Several studies have suggested benefits of this intervention.

The aim of this study is to determine the biochemical effects of treatment with vitamin D in patients with PHPT.

Clinical records of 70 patients with PHPT were retrospectively analyzed. Included patients had VD < 30 ng/ml, without prior supplement of VD >800UI/daily and were treated with higher dose VD during follow-up. Patients with additional conditions or treatments affecting calcium homeostasis were excluded, resulting in a study group of 26 patients, 25 female, mean age 56 years (range 25-79). Eleven patients were treated with cholecalciferol 100000UI periodically (estimated daily dose 3200±500) and 15 with ergocalciferol weekly (estimated daily dose 4500±2200). Evaluations were performed baseline and after 1 to 12 months of VD treatment, comparisons for dependent samples were made with appropiate tests for the distribution of the variables. PTH was expressed as percentage of the upper normal limit (%UNL), to allow comparisons. No significant changes with VD supplementation were seen in Ionized calcium (baseline vs under treatment 5,88±0,06 mg/dl vs 5,96±032 mg/dl; p=0.058), total calcium (10,5±0,6 mg/dl vs 10,6±0,9; p=0.26), phosphorus (3.37±0.6 vs 3,2±0.5, p=0,11) or PTH (median 158 %UNL vs 140 %UNL, p=0.119). Calciuria increased significantly with VD treatment (median 259 mg/24hs vs 310 mg/24 hs), without direct correlation with variations of calcemia, PTH or VD. Natriuria was evaluated in 10 patients and showed correlation with rises in calciuria (R2 57,4%, P 0,029). Alkaline Phosphatase (ALP) showed significant reduction with VD (282±100 IU/L vs 247±62 IU/L, p 0.016). When stratifying patient according to VD level obtained with treatment (20-30 ng/dl vs >30 ng/dl) no difference between groups was observed, with the exception of a tendence of ALP to fall when higher levels of VD were reached.

Supplementation with VD did not rise significantly calcemia in patients with PHPT and VDD. VD lowered ALP, probably by reduction of bone remodelling and improvement of intestinal calcium absorption. It is advisable to supplement VD in these patients, given the lack of risks and potential benefits of the treatment.


Nothing to Disclose: BMP, MPP, RS, GRC, MP, SVK, HHS

26266 15.0000 FRI 340 A Effects of Vitamin D Supplementation in Primary Hyperparathyroidism 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 324-348 7679 1:15:00 PM Parathyroid Disorders and Metabolic Bone Disease (posters) Poster

Georgios Kontogeorgos*1, Christine Laine1, Lennart X Welin2, Michael Fu3, Per-Olof Hansson4 and Kerstin Landin-Wilhelmsen1
1Section for Endocrinology, Institution of Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden, 2Department of Medicine, Gothenburg, Sweden, 3Dept of Cardiology, Gothenburg, Sweden, 4Dept of Medicine, Gothenburg, Sweden


Objective: Normocalcemic, vitamin D sufficient hyperparathyroidism (nHPT) in men and women has been shown to have a benign course1. The aim was to study the prevalence and morbidity of nHPT and other parathyroid hormone (PTH) and calcium aberrations in 50-year-old men during 20 years´ follow-up.

Design: Cross-sectional and retrospective study at the Sahlgrenska University Hospital, Gothenburg, Sweden.

Subjects: A random population of 797 men born in 1943, was studied in 1993 (of whom 47% were vitamin D sufficient).

Measurements: A serum-(S) PTH >6.9 pmol/l was considered as HPT, S-calcium 2.15-2.49 mmol/l as normocalcemia, and S-25(OH)D >50 nmol/l as vitamin D sufficiency. Data on fractures, stroke and myocardial infarction were retrieved until 2013, yielding a 20-year follow-up.

Results: The prevalence of primary (p) HPT was 0.4% (one man with kidney stones; underwent parathyroid surgery), secondary HPT 0.3%, nHPT 2.7%, hypercalcemia with normal S-PTH 0.3%,  hypocalcemia with normal S-PTH 3.1%, and HPT with vitamin D insufficiency 5.5% in 1993 (age 50). Hypertension developed in all with pHPT. Hypertension was not more common in nHPT than in men without calcium/PTH aberrations during follow-up. No increase in fractures, diabetes, kidney stones (except for the man with pHPT), myocardial infarction, stroke, cancer or death was seen at follow-up.

Conclusions: Primary HPT was rare (0.4%), but normocalcemic, vitamin D sufficient HPT was common, 2.7%, in men aged 50 years. No increase in hard end points was seen in men with, compared with men without, calcium/PTH aberrations over a 20-year period.


Nothing to Disclose: GK, CL, LXW, MF, POH, KL

26814 16.0000 FRI 341 A Hyperparathyrodisim in Men - Low Morbidity and Mortality during 20 Years´Follow-up 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 324-348 7679 1:15:00 PM Parathyroid Disorders and Metabolic Bone Disease (posters) Poster

Laura J Reid*, Balakumar Muthukrishnan, Andrew Ditchfield, Andrew Brodie and Fraser W Gibb
Edinburgh Centre for Endocrinology & Diabetes, Edinburgh, United Kingdom


Background/Aim:  Primary hyperparathyroidism is an increasingly common condition, which is often managed conservatively, particularly in older patients with relatively modest hypercalcemia.  Recent evidence suggests that increased PTH may be associated with higher mortality.  We sought to characterize the diagnostic features and management of patients referred for evaluation of primary hyperparathyroidism in a single endocrine centre.

Methods:  We undertook a retrospective review of the diagnosis and management of all patients with primary hyperparathyroidism attending an endocrine clinic, within a large university teaching hospital, between 2009 and 2013.  Clinical, biochemical and imaging data were extracted from electronic patient records.  Data are median (inter-quartile range).

Results: The majority of patients were women (198/241), presenting at an older age than men (70 [49 – 79] vs. 64 years [52 – 73], p = 0.013).  Median calcium at presentation was 2.75 mM (2.67 – 2.87) and PTH 12.4 pM (9.3 – 18.9).  Neck ultrasound identified the source in 101/142 patients, sestamibi identified a further 12 and CT/SPECT a further 4.  Abdominal imaging in 87 patients revealed nephrolithiasis in 14 (16%). Of 132 assessed, 42.4% had osteoporosis and a further 40.2% osteopenia.  Vitamin D deficiency was present in 30.9% and was associated with significantly higher PTH (14.7 [12.2 – 21.3] vs. 9.7 nM [7.8 – 15.8], p = 0.003) than replete individuals (25.8%).  Vitamin D replacement (18.7%) was not associated with significant change in calcium, in conservatively managed patients.

In total, 42.3% (n = 102) proceeded to surgery:  a significantly younger cohort (62 [50 – 72] vs. 73 years [64 – 80]), with higher calcium (2.81 [2.70 – 2.95] vs. 2.72 mM [2.63 – 2.82]) and PTH (15.9 [10.4 – 22.1] vs 11.6 pM [8.9 – 15.1]); all p < 0.001.  Surgical cure was achieved in 92.1%. At least one neck imaging modality had been positive in patients with persistent disease post-surgery.  25.3% of all patients presented with a calcium >0.25 mM above the upper limit of normal (2.85 mM). Where surgery was not performed, median calcium fell during two years of observation, with a greater fall observed in those with higher calcium at presentation (-0.11 vs. -0.05, p = 0.003). After 2 years of follow up, no patients with initial serum calcium <2.85 mM (n = 93) had risen above this threshold and 10/12 patients, above 2.85 mM at diagnosis, fell below the threshold at 2 years.

Conclusions:  These data provide reassurance that conservative management (including vitamin D replacement) is not associated with a significant risk of worsening hypercalcaemia over a 2-year period.  When complete, our cohort of over 400 patients should provide further information on the prevalence of complications and impact of newer imaging modalities upon adenoma localization and cure rates.  Large prospective databases present an opportunity to optimize care in this common condition.


Nothing to Disclose: LJR, BM, AD, AB, FWG

26867 17.0000 FRI 342 A Extensive Clinical Experience in the Investigation and Management of Primary Hyperparathyroidism 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 324-348 7679 1:15:00 PM Parathyroid Disorders and Metabolic Bone Disease (posters) Poster

Danica Maria Vodopivec*, Angelica Maria Silva, Kelly Schwarz, Ioannis Christakis, Ashley Stewart, Mimi I-Nan Hu and Nancy D Perrier
The University of Texas MD Anderson Cancer Center, Houston, TX


Introduction: Sporadic primary hyperparathyroidism (sPHPT) affects up to 1% of the adult population, being more commonly detected in women, especially post-menopausal (post-MP). Surgery for sPHPT is associated with an increase in post-operative BMD, but there is no published literature on gender differences regarding BMD recovery. We aimed to investigate differences in BMD changes after PTX between men and women (pre-MP and post-MP) with sPHPT.

Methodology: Retrospective study of all adult patients who underwent PTX in our Institution (1990-2013). Patients were included if they had both pre- and post-operative DXA scans performed in our institution (biggest limitation for recruitment since most of our patients were referred with their initial DXA scan done at an outside institution), no recurrent or persistent disease post operatively, and did not receive any medical treatment interfering with bone or calcium metabolism.

The percentage change in BMD (gr/cm2) and T-scores were analyzed at baseline (pre-operative) and 12±6 months after surgery in the lumbar spine (LS), femoral neck (FN), total hip (TH) and distal one-third of the radius (D 1/3 R). Values were then compared between men and women (pre-MP and post-MP). p value < 0.05 was considered statistically significant.

Results: 1,467 patients underwent PTX and of these 105 patients met inclusion criteria. There were 72 women (68.6%; 13 pre-MP, 58 post-MP and 1 patient with unknown menopausal status) and 33 men (31.4%). The average age at PTX for men and women was 58±2 and 62±1 years old, respectively (p= 0.1707). The average time period between PHPT diagnosis and PTX for men and women was 5±1 and 11±2 months (p= 0.1322), respectively. There was no significant change in the BMD of D 1/3 R between study groups, thus this anatomical site is no further mentioned in the study.

There was no difference in initial severity of bone disease between post-MP and men in all 3 sites, and pre-MP had higher initial T-score values than men in the FN (p=<0.0001) and TH (p=0.0036). Pre-MP also had higher pre-operative T-scores than post-MP in all 3 sites (LS: p=0.0082, FN: p=<0.0001, TH: p=0.0015).

One year post-PTX men had 1.6% greater increase in their LS BMD (gr/cm2) when compared to females (p=0.0352).   Post-MP recovered their BMD values (gr/cm2) less than men in all three sites, and this was found to be significant in the FN and TH where men had 2.3% (p=0.0443) and 0.2% (p=0.0443) greater increase, respectively.

Conclusion: The significant increase in the BMD values (gr/cm2) of men after PTX should be reiterated as a potential surgical benefit to cure in this gender. Unfortunately, because sPHPT is more common in women, there is an epidemiologic tendency to underdiagnose sPHPT in men and to underutilize BMD in this gender because they are not considered as having clinically significant bone loss.


Nothing to Disclose: DMV, AMS, KS, IC, AS, MINH, NDP

27530 18.0000 FRI 343 A Bone Mineral Density Screening Importance in Men with Sporadic Primary Hyperparathyroidism 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 324-348 7679 1:15:00 PM Parathyroid Disorders and Metabolic Bone Disease (posters) Poster

Shirin Kalyan*1, Millan Patel2, Elaine Kingwell2, Helene Cote3, Danmei Lui3 and Jerilynn C Prior1
1University of British Columbia, Vancouver, BC, Canada, 2University of British Columbia, Vancouver, BC, 3University of British Columbia


Background: Polycystic ovary syndrome (PCOS) is a common, complex endocrine disorder characterized by androgen excess, menstrual cycle irregularities, ovulatory disturbances and abdominal adiposity. Although women with PCOS often have health issues related to insulin resistance and central obesity, the prevailing view is that they have normal or stronger bones and fewer fractures because of heavier weight and higher androgen levels. However, findings from a cohort-control study with 315 premenopausal women did not concur with this theory: rather than being protected from bone fractures, women with PCOS had an increased rate of multiple (≥2) fractures (8% of women with PCOS vs. 2.8% of control women).

Hypothesis: Inflammation and oxidative stress, which are often present in women with PCOS, may compromise bone quality.

Methods: In a cross-sectional convenience sample, we investigated total hip areal bone mineral density (BMD) by dual-energy X-ray absorptiometry and radius strength-strain index (SSI) by peripheral quantitative computed tomography in 22 premenopausal women with PCOS (mean age 40.3±3.4 years [y]) and 39 controls (mean age 42.5±4.2 y). We explored relationships among bone parameters, inflammation (C-reactive protein [CRP]/albumin ratio), oxidative stress (leukocyte telomere length and DNA damage), combined hormonal contraceptives (CHC) use and anthropometric characteristics.

Results: Women with PCOS had significantly greater BMI (29.7±7.9 kg/m2) compared to controls (25.1±4.6 kg/m2), p=0.003. There was no difference in CHC usage (18% in women with PCOS vs. 15% controls). A diagnosis of PCOS was a negative predictor (beta=-0.25, p=0.022) of total hip BMD in a linear regression model that was adjusted for current body weight. Increased subclinical inflammation by CRP/albumin ratio was associated with decreased radius SSI in women with PCOS (R2= 0.25, p=0.018), but not in controls. The CRP/albumin ratio was positively predicted by an increased waist-to-height ratio and current CHC use. There was no significant association between any of the bone parameters and our oxidative stress measures.

Conclusions: These data suggest that the presence of subclinical inflammation may compromise bone quality in women with PCOS. Greater consideration of CHC effects on subclinical inflammation and bone homeostasis in treatment for PCOS may be warranted. Further research on the physiology of bone in women with PCOS is necessary.


Nothing to Disclose: SK, MP, EK, HC, DL, JCP

26272 19.0000 FRI 345 A Does Subclinical Inflammation Compromise Bone Quality in Women with Polycystic Ovary Syndrome? 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 324-348 7679 1:15:00 PM Parathyroid Disorders and Metabolic Bone Disease (posters) Poster

Joely A. Straseski*1 and Mark M Kushnir2
1University of Utah & ARUP Laboratories, Salt Lake City, UT, 2ARUP Laboratories, Salt Lake City, UT


Parathyroid hormone related protein (PTHrP) is involved in intracellular calcium regulation and may be measured in patients suspected of hypercalcemia of malignancy. PTHrP and its gene are also known to be expressed in number of normal cells and tissues. We previously developed a method for the measurement of PTHrP in plasma using LC-MS/MS and established reference intervals in healthy adults. Measurable concentrations of PTHrP were observed in plasma samples of all participants, suggesting that PTHrP is present in circulation in health. The aim of this study was to evaluate associations between PTHrP concentrations and age in men and women and to evaluate between-gender differences.

The LC-MS/MS method was fully validated according to CLSI guidelines and is in routine use in a clinical laboratory. Briefly, stable isotope-labeled internal standard was added to samples and PTHrP was enriched using anti-PTHrP antibody conjugated to magnetic beads, digested with trypsin and samples were analyzed by LC-MS/MS. The lower limit of quantification and upper limit of linearity of the assay were 0.3 and 1100 pmol/L, respectively. Total imprecision of the method was < 10%. Specificity of the measurements was confirmed by monitoring two mass transitions of PTHrP and the internal standard. Using this method we analyzed 284 plasma samples collected from adults: 132 men (age 18-81 y, mean 40 y) and 152 women (age 18-84 y, mean 41 y). Differences between groups were evaluated using nonparametric statistics.

Overall, significantly higher PTHrP concentrations were observed in women compared to men (p<0.0001). In women, the highest concentrations were observed in the 21-30 y group, concentrations were minimal in the 41-50 y group and then rose progressively with age. Statistically significant differences in concentration were observed between the age groups of women 18-30 y and 31-40 y (p<0.0078); 18-40 y and 41-50 y (p<0.0405); and 41-50 y and 51-84 y (p=0.0001). Notably, statistically significant higher concentrations were observed in women of post-menopausal age (>50 y vs. <50 y, p=0.0009). In men, the lowest concentrations were observed between the ages of 21 and 50 y.  Lower concentrations were observed in the age group 51-60 y compared to 61-81 y (p=0.019). Higher concentrations were observed in women than in men in the age groups of 21-30 y (p=0.0017), and 51-84 y (p=0.0054).

In general, concentrations of PTHrP were higher in women than in men and were statistically significantly higher in older individuals. The highest concentrations of PTHrP were observed in women 21-30 y and > 51 y, and in men >71 y. This gender and age distribution overlaps with many age-related disease processes, including osteoporosis, and raises questions regarding PTHrP’s involvement in disorders of calcium regulation associated with age.


Nothing to Disclose: JAS, MMK

25813 20.0000 FRI 346 A Age and Gender-Related Variation in Concentrations of Parathyroid Hormone-Related Protein Measured By LC-MS/MS 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 324-348 7679 1:15:00 PM Parathyroid Disorders and Metabolic Bone Disease (posters) Poster

Karin Amrein*1, Verena Schwetz2, Nicolas Verheyen1, Astrid Fahrleitner-Pammer3, Christian Schnedl4, Thomas R Pieber5 and Harald Dobnig6
1Medical University of Graz, Graz, Austria, 2Medical University of Graz, Steiermark, Austria, 3Medical University, Graz, Austria, 4Department of Radiology, Graz, Austria, 5Medical University of Graz, Austria, 6Schilddruesen|Endokrinologie|Institut, Graz, Austria


Introduction: Fibroblast growth factor 23 (FGF23) is a phosphaturic hormone and regulator of vitamin D metabolism that seems to be a useful biomarker for the identification of high-risk patients in chronic but also acute disease. It is produced mainly in bone, although the failing heart has recently been suggested to be a source of circulating FGF23via oncostatin M as well. FGF23 rises rapidly in acute and chronic kidney disease and is strongly and independently associated with excess morbidity and mortality. Emerging data suggest that FGF23 is also predictive of poor outcomes in different scenarios of acute illness including cardiogenic shock and cardiac surgery.

Methods: In this posthoc analysis in the placebo group of the VITdAL-ICU study, we analysed FGF23 levels in a subgroup of adult medical/surgical critically ill patients with vitamin D deficiency (25-hydroxyvitamin D equal or lower to 20ng/ml) at baseline (n= 65) and day 7 (n=49). C-terminal FGF-23 was measured in one batch from frozen stored plasma samples by enzyme-linked immunosorbent assay (Immunotopics, San Clemente, CA, USA, upper detection limit after 1:10 dilution 14,000 RU/ml).

Results: Overall, median FGF23 levels were 242 RU/ml at day 0 and 212 RU/ml at day 7 (R=0.67, p<0.01). Patients who died in the hospital (n=19, 29%) had almost 10-fold and significantly higher baseline FGF23 levels than survivors (1500 vs. 170 RU/ml, P<0.01). There was a strong correlation between FGF23 and kidney function, the Simplified Acute Physiology Score II, the Charlson comorbidity index and 1,25-dihydroxyvitamin D (P<0.01), but not with other biochemical markers including phosphate and calcium.

Discussion: FGF23 is an interesting novel biomarker that is a strong independent indicator of poor outcomes in chronic kidney disease. Its role in acute disease is less studied, but appears to be similarly predictive for adverse outcomes which is supported by our data in critical illness. It remains unclear if FGF23 is only a marker or also a contributor of adverse outcomes and if dietary and/or pharmacological interventions to reduce FGF23 concentrations would relate to clinical benefit.


Nothing to Disclose: KA, VS, NV, AF, CS, TRP, HD

24343 21.0000 FRI 347 A FGF-23 As a Novel Predictor of Hospital Mortality in Critically Ill Patients 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 324-348 7679 1:15:00 PM Parathyroid Disorders and Metabolic Bone Disease (posters) Poster

Margaret Wolfe-Wylie*, Marialena Mouzaki, Etienne B Sochett and Jennifer Jean Harrington
The Hospital for Sick Children, Toronto, ON, Canada


Background: Proton pump inhibitors (PPIs) are associated with a modest increased fracture risk in adults. The underlying mechanism is unknown, but has been postulated to involve decreased calcium absorption or altered osteoclast function. The effect of PPIs on phosphate metabolism has not been previously well investigated. We present a case-series of five pediatric patients who developed hypophosphatemic rickets on chronic high dose PPI therapy. We propose that decreased dietary phosphate absorption secondary to increased gastric pH may be a previously unrecognized cause for bone fragility in children on PPIs.

Clinical Cases: Five children (median age 2.9 [0.9 to 3.8] years), all with a history of developmental delay, gastrostomy-tube feeds and severe gastroesophageal reflux (GERD) requiring chronic high dose PPI therapy (mean omeprazole dose 3.6±0.7 mg/kg/day for a mean of 2.4±1.3 years), were diagnosed with hypophosphatemic rickets (1 presented with bone pain, 2 with fractures, 2 were asymptomatic with lab abnormalities). All had radiological evidence of rickets. On further investigation, common results included: hypophosphatemia (0.61-0.85 mmol/L; normal range [NR] 1.36-2.17), normocalcemia, elevated alkaline phosphatase (611-1192 U/L; NR 185-520), low/normal PTH (7-36 ng/L; NR 12-78), normal 25-hydroxyvitamin D (>70 nmol/L), elevated 1,25 dihydroxy vitamin D (261-1146 pmol/L; NR 39-193) with normal renal function. Renal tubular maximum reabsorption rate of phosphate/glomerular filtration rates (TmP/GFR) were available in 4 patients, all of which were normal. Cumulatively these results excluded excess renal phosphate losses (including via excess PTH or fibroblast growth factor-23), and instead suggest decreased intestinal phosphate absorption as the probable underlying aetiology for the hypophosphatemia. Four of the patients were commenced on phosphate supplementation with subsequent significant improvement in the rachitic and biochemical (phosphate and ALP) changes. The remaining patient had complete resolution of the hypophosphatemic rickets without phosphate supplementation, after being weaned off of the PPI.

Conclusion: This is the first case series of hypophosphatemic rickets in association with high dose PPI administration. In particular, the case of complete biochemical and radiological resolution of the hypophosphatemic rickets with weaning of PPI alone, supports the hypothesis that decreased intestinal phosphate absorption from PPI use was the underlying aetiology.   This case-series provides justification for a formal study of the mechanism, prevalence, and severity of hypophosphatemia and its subsequent effects on bone mineralization in the setting of chronic high dose PPI therapy in children.


Nothing to Disclose: MW, MM, EBS, JJH

25774 22.0000 FRI 348 A Chronic High Dose Proton-Pump Inhibitors As a Cause of Hypophosphatemic Rickets 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 324-348 7679 1:15:00 PM Parathyroid Disorders and Metabolic Bone Disease (posters) Poster

Katherine J Motyl*1, Clifford J Rosen2 and Karen L Houseknecht3
1Maine Medical Center, Scarborough, ME, 2Maine Medical Center Research Institute, Scarborough, ME, 3University of New England, Biddeford, ME


Atypical antipsychotic (AA) drugs are among the top selling in the United States. However, AA drugs have a variety of metabolic side effects, including hyperglycemia, obesity and cardiac arrest.  AA drugs also increase fall and fracture risk and reduce bone mineral density (BMD), the latter occurring even in adolescents. We have demonstrated trabecular (Tb) bone loss from daily oral 0.75 mg/kg risperidone (RIS) in 8 wk, female C57BL/6J (B6) mice, a dose that achieves clinically relevant plasma levels of RIS and its active metabolite, 9-OH RIS. We have also shown that bone loss may be through the sympathetic nervous system (SNS). However, the prevailing mechanism of bone loss in the literature is hyperprolactinemia-induced hypogonadism due to RIS-mediated inhibition of dopamine receptor (D2R) signaling. We hypothesized that if hypogonadism was the only cause of bone loss after RIS treatment, then mice that were ovariectomized (OVX) and RIS treated would not lose any more bone than that which is lost by OVX alone. Mice were sham operated (SH) or OVX at 5 wks of age. At 8 wks, a subset of each group was given vehicle (VEH) or 0.75 mg/kg RIS daily, such that we had a total of 4 groups (SH+VEH, SH+RIS, OVX+VEH and OVX+RIS, N=15 each). Consistent with our previous findings, 8 wks of RIS in SH mice caused an 18% reduction of Tb bone volume fraction (BV/TV, p=0.02), as well as significantly reduced connectivity density (p<0.01) and Tb number (p<0.01) compared to SH+VEH. Also as expected, Tb BV/TV was reduced by 55% in OVX+VEH mice compared to SH+VEH (p<0.01). Despite the dramatic loss of bone induced by OVX, OVX+RIS mice lost even more Tb bone such that their BV/TV was 30% lower than that of OVX+VEH (p=0.04). Furthermore, there was not a significant interaction between the main effects of OVX and RIS treatment by 2-way ANOVA (p=0.61), suggesting estrogen-deficiency did not play a role in RIS-mediated bone loss. RIS did not affect cortical bone in SH or OVX mice (main effect p=0.36), but all OVX mice had significantly lower cortical BV/TV (main effect p<0.01). Consistent with our previous findings, RIS did not alter body mass in SH mice (p=0.75) and OVX significantly increased body mass in VEH treated mice (p<0.01). However, RIS attenuated weight gain in OVX (p<0.01, interaction p<0.01). DXA analyses revealed that changes in body mass from both RIS and OVX were entirely due to changes in fat mass. Collectively, these findings suggest the adipose tissue, but not bone, consequences of RIS are dependent upon estrogen status. In conclusion, hyperprolactinemia-induced hypogonadism cannot explain bone loss in a mouse model of clinically relevant AA treatment. RIS-mediated bone loss is likely multi-modal, with the SNS and direct RIS-bone interactions playing important roles that need to be considered when designing therapies and making treatment decisions.


Nothing to Disclose: KJM, CJR, KLH

27237 2.0000 FRI 350 A Hyperprolactinemia-Induced Hypogonadism Does Not Explain Bone Loss from the Atypical Antipsychotic Risperidone 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 349-367 7685 1:15:00 PM Osteoporosis - Risks and Treatments (posters) Poster

Kriti Gupta*1, Preethi Kadambi1 and Eric S Nylen2
1GWU, Washington, DC, 2VA Medical Center, Washington, D.C.


Introduction: It is well established that there is an inverse relationship between bone mineral density and atherosclerosis (Becker et al 1972).  Moreover, bone remodeling agents may benefit both bone and vasculature.  Denosumab is a monoclonal antibody against RANKL, and is the agent of choice for osteoporosis in patients with chronic kidney disease (CKD).  Its impact, however, on kidney function has not been well studied. 

Objectives: The primary objective was to assess kidney function following Denosumab injections in elderly osteoporotic subjects.

Methods: We identified 21 patients (mean age 78.8±10.2 years) who had received Denosumab injections at the VA Medical Center, Washington, DC between 2012 and 2015.   Kidney function was calculated using the CKD-EPI (Chronic Kidney Disease Epidemiology Collaboration) equation. Comparison between kidney function of patients before and after Denosumab injections was made using a paired t-test.

Results: Of the 21 patients, 11 were Caucasian (52%), 9 were African-American (43%), and 1 was Filipino (5%).  Of the 21 patients, 5 were female (24%) and 16 were male (76%).  The average eGFR in the pre-Denosumab group was 59.86±30.61 mL/min/1.7m2 and in the post-Denosumab group eGFR was 66.67±31.52 mL/min/1.7m2.  Kidney function in the post-Denosumab group improved by 11.36% (p<0.004).   In the pre-Denosumab group, there were 15 patients (71.4%) who were in CKD Stage 3, of which 7 patients improved to CKD Stage 2 after Denosumab.

Conclusions: In this pilot study of patients with high risk for CVD (i.e., elderly, osteoporotic with CKD) Denosumab administration resulted in an increase in eGFR by 11.4% and improved the CKD stage in 46.7% of subjects.   Thus, osteoporotic treatment may modify vascular risk factors for CVD by altering the progression of CKD.    


Nothing to Disclose: KG, PK, ESN

25871 4.0000 FRI 352 A Impact of Denosumab on Renal Function in High Risk CVD Subjects 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 349-367 7685 1:15:00 PM Osteoporosis - Risks and Treatments (posters) Poster

Javier Mauricio Farias*1, Cesar Bogado2, Maria Belen Zanchetta2, Fabio Masari3, Mariana Papouchado4, Marcelo Criscuolo4, Roberto A Diez4 and Jose R Zanchetta2
1Sanatorio Guemes, Ciudad auronoma de Buenos Aires, Argentina, 2IDIM CR, Buenos Aires, Argentina, 3IDIM CR, Argentina, 4Biosidus SA


A randomized single blind trial in a single osteoporosis reference center was designed to compare the efficacy and safety of Osteofortil® (Biosidus, Argentina) a new pharmaceutical product containing teriparatide, and the reference product Forteo® (Eli-Lilly, France) in posmenopausal women with Osteoporosis. Teriparatide dose was 20 µg per day by subcutaneous injection. Inclusion criteria: age between 50 and 81 years, with osteoporosis defined by mineral bone density (MBD) on lumbar spine (LS) < -2.5 (Tscore) or lumbar fracture with MBD <-2 on LS, femoral neck or total hip. An ethical committee and the national drug regulatory agency of Argentina (ANMAT) approved the trial, also registered in clinicaltrials.gov (NCT # 01945788). A comparative six-month initial phase was followed by an extension phase in which all patients were invited to receive Osteofortil® for 6 months. Data was analyzed using the STATA 12 package (Stata Corp). A total of 192 patients were enrolled and 100 met inclusion and exclusion criteria, 95 and 72 completed the comparative and the extension phase, respectively. Osteocalcin levels (O), N-terminal propeptide of procollagen type 1 (P1NP), C-terminal cross-linked telopeptide of type I collagen (CTX) were measured by Roche E411 ECLIA Electrochemiluminescence, at baseline and months 1, 3, 6, 9 and 12. MBD was assessed with DEXA (dual-energy x-ray absorptiometry) by Lunar Prodigy ™, GE Healthcare, Madison, WI, USA and by quantitative computed tomography (QCT), GE, Brightspeed Excel Select, at baseline, 6 and 12 months. The mean bone density by T-score was -2.9 and age was 65 years with no differences in age at menopause, previous use of bisphosphonates, vitamin D and the presence of fractures at baseline between both groups. At 6 months Osteofortil® n = 48 increased O levels 125%, P1NP 160% CTX 162%, whereas for reference product was 124%, 164% and 186 % respectively. No differences between groups were found. Tscore at 6 and 12 months was -2.6±0.5 and -2.5±0.6 with Osteofortil and -2.59±0.48, -2.5±1.2 with reference product. The increase in MBD in LS by DEXA at 6 and 12 months was 4.4%, 6.58% and 6.3%, 7.6% with innovator and reference product, and 15% and 17 % by QCT at 6 months, respectively. Not significant differences was found. Similar rates of hypercalciuria and low vitamin D was found between groups. Conclusions: Both products resulted in similar increase in bone markers and mineral bone density in the lumbar spine. Both depicted similar safety


Nothing to Disclose: JMF, CB, MBZ, FM, MP, MC, RAD, JRZ

26985 5.0000 FRI 353 A Clinical Comparison of Two Teriparatide Formulations: Osteofortil® and Forteo® 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 349-367 7685 1:15:00 PM Osteoporosis - Risks and Treatments (posters) Poster

Daichi Miyaoka*, Yasuo Imanishi, Masaya Ohara, Noriyuki Hayashi, Yuki Nagata, Shinsuke Yamada and Masaaki Inaba
Osaka City University Graduate School of Medicine, Osaka, Japan


Trabecular bone score (TBS), a textural index that evaluates pixel gray-level variations in the lumbar spine (LS) dual energy x-ray absorptiometry (DXA) image, is a new surrogate for trabecular bone microarchitecture assessment which is associated with fracture risk independently of bone mineral density (BMD) (1). Although teriparatide (TPTD) increased LS-BMD as well as TBS in 2 years (2), the precise time-course of these parameters was not reported. In this study, we attempted to determine the changes in LS-BMD and TBS in osteoporotic patients treated with TPTD followed by minodronate (MINO).

Osteoporotic patients, with low LS-BMD (T-score< -2.5) and/or at least one vertebral fracture, were treated with TPTD (20 μg/day) (n= 29; age, 67.8 ± 12.6 years; BMI, 21.4 ± 2.60 kg/m2; LS-BMD, 0.687 ± 0.146 g/cm2; TBS: 1.197 ± 0.107, mean ± SD) for 12-24 months, followed by oral MINO (50 mg/4 weeks) administration. LS-BMD (L1-4, QDR-2000, Hologic Inc., MA) and TBS (iNsight software, Med-Imaps, France) were measured at 0, 3, 6, 12 months of TPTD therapy, and 12 months after the initiation of MINO.  The study was approved by the institutional ethics committees and was conducted in accordance with the principles of the Declaration of Helsinki.

The correlation of baseline TBS and LS-BMD was borderline significant (r= 0.3629, P= 0.053). The increments of LS-BMD were significant at 6 months (3.88 ± 0.97%, P< 0.001) and kept increasing until 12 months (6.70 ± 1.21%, P< 0.001). The increments of TBS were already significant at 3 months (3.05 ± 1.00%; P= 0.029) and were stable until 12 months. Twelve months MINO therapy after TPTD maintained both BMD (0.58 ± 1.07%, P= 0.592) and TBS (0.40 ± 0.87%, P=0 .647).

Different time-courses between LS-BMD and TBS suggested that these two parameters measured different responses of bone to TPTD therapy. Trabecular microarchitecture seemed to improve faster than mineralization. Bisphosphonate following TPTD was beneficial in maintaining TBS as well as BMD.


Nothing to Disclose: DM, YI, MO, NH, YN, SY, MI

25072 6.0000 FRI 354 A Effects of Teriparatide and Sequential Minodronate Therapies on Lumbar Bone Mineral Density and Microarchitecture Assessed By the Trabecular Bone Score in Japanese Patients with Osteoporosis 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 349-367 7685 1:15:00 PM Osteoporosis - Risks and Treatments (posters) Poster

Penelope Trimpou*, Georgios Kontogeorgos, Christine Laine and Kerstin Landin-Wilhelmsen
Section for Endocrinology, Institution of Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden


Background: Osteoporosis and fractures are common in the population, especially in women. Long-term follow up studies of anabolic treatment in this metabolic bone disease are sparse.

Objective: The aim was to compare effects of two different anabolic hormone treatments for established osteoporosis; teriparatide (Forsteo®) and growth hormone (GH) (Genotropin®) with regards to bone mineral density (BMD) and fracture outcome after 3 years.

Subjects and Methods: Teriparatide 20 µg was given subcutaneously to 50 subjects (44 women and 6 men; mean age 66+12.9 years) during 18 months followed by bisphosphonates, in 62%, up to 36 months. All treated subjects had established osteoporosis and at least one vertebral compression fracture. BMD measured by DXA and fracture rates were monitored. Results were compared with 55 women of similar age and condition who received subcutaneous GH, either 1.0 U or 2.5 U daily during 36 months1. The subjects had postmenopausal osteoporosis and 45% had fractured before treatment. All subjects received 1000 mg calcium and 800 U vitamin D throughout the study period. Comparison was made with register data from a random population sample of 226 age-matched women in parallel and 20% had fractured at start, the WHO MONICA study, Gothenburg, Sweden.

Results:  BMD increased with teriparatide similarly to GH during 18 months. Subsequent treatment with bisphosphonates gave similar results, 3% increase of total body BMD, 8% at the lumbar spine and 5% at the femoral neck, as in the GH-treated subjects after 36 months. No effect was seen on lean body mass of teriparatide but increased with the high dose of GH. Fractures occurred in 6% of the teriparatide group, 0% in the GH treated group and in 4% of the population sample where supplementation and bone specific agents were rarely used, <1%. Four women discontinued teriparatide treatment due to side effects (pain, vertigo) while all subjects on GH completed the regimen.

Conclusion: Anabolic treatment with teriparatide (PTH) followed by bisphosphonates, was as effective as GH treatment for 3 years in established osteoporosis regarding BMD. Fractures still occurred in the severely affected group while on treatment. Bone specific treatment in subjects with fractures in the population was negligible.


Nothing to Disclose: PT, GK, CL, KL

27404 7.0000 FRI 355 A Comparison Between Teriparatide (PTH) and Growth Hormone (GH) Treatment during 3 Years in Established Osteoporosis 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 349-367 7685 1:15:00 PM Osteoporosis - Risks and Treatments (posters) Poster

Melanie Schorr*1, Dean A. Marengi2, Reitumetse L. Pulumo2, Kamryn T. Eddy1, Anne Klibanski1, Karen K. Miller1 and Elizabeth A. Lawson1
1Massachusetts General Hospital/Harvard Medical School, Boston, MA, 2Massachusetts General Hospital, Boston, MA


Oxytocin (OXT) is an anorexigenic hypothalamic hormone, secreted into the peripheral circulation via the posterior pituitary, with important metabolic effects.  We previously hypothesized that basal OXT levels reflect energy availability and found basal serum OXT levels to be lower in anorexia nervosa (AN, a state of energy deficit) than healthy controls (HC). However, reports are conflicting regarding OXT levels in overweight/obesity (OB, a state of energy excess). Preclinical studies indicate that OXT is anabolic to bone. Low overnight serum OXT levels (pooled q 20 min) are associated with reduced PA and lateral spine bone density (BMD) in AN and impaired bone microarchitecture in amenorrheic athletes, but OXT’s association with BMD in OB, and hip geometry across the weight spectrum, is unknown. We hypothesized a positive linear relationship between overnight serum OXT levels and BMD as well as hip structural analysis (HSA) parameters, both of which predict fracture risk.

In a cross-sectional study, we evaluated 59 women, 18-45 y: eumenorrheic OB (N=19), amenorrheic AN (N=16), eumenorrheic HC (N=24). Serum was sampled q 20 min 8 PM – 8 AM and pooled for an integrated measure of overnight OXT levels. Body composition, BMD and HSA were measured by dual x-ray absorptiometry.  Mean age was similar among groups (28.3 ± 7.7 y, mean ± SEM). Per design, BMI and total fat differed among groups (OB- 32.4 ± 1.4 kg/m2, 35.2 ± 2.6 kg; AN- 18.4 ± 0.2 kg/m2, 9.6 ± 0.6 kg; HC- 22.4 ± 0.3 kg/m2, 16.4 ± 0.7 kg) (p<0.005). OXT levels were lowest in AN and highest in OB (AN 320.3 ± 41.3 pg/mL, HC 442.4 ± 56.0 pg/mL, OB 533.1 ± 44.0 pg/mL) (AN vs HC p=0.03, AN vs OB p=0.002, OB vs HC p=0.04). There was a positive linear relationship between OXT and 1) BMI (R=0.51, p<0.0001), total fat (R=0.47, p<0.0001), visceral fat (R=0.47, p=0.0002) and subcutaneous fat (R=0.47, p=0.0002); and 2) BMD Z-scores at the PA spine (R=0.45, p=0.0004), lateral spine (R=0.46, p=0.001) and total hip (R=0.33, p=0.01). After controlling for BMI, the association between OXT and PA and lateral spine Z-scores trended toward significance (p≤0.10). OXT was positively associated with narrow-neck (NN) cortical thickness (cort thick) (R=0.40, p=0.002), NN cross-sectional area (CSA) (R=0.28, p=0.03), inter-trochanteric (IT) cort thick (R=0.30, p=0.02) and IT CSA (R=0.28, p=0.03); and negatively associated with NN buckling ratio (BR) (R=0.35, p=0.007) and IT BR (R=0.32, p=0.01) (lower BR denotes higher strength). The relationships with NN cort thick and NN BR remained significant after controlling for BMI (p<0.05).

In summary, we found that nocturnal OXT levels were higher in OB and lower in AN compared to HC and positively associated with BMI, fat mass and bone parameters across the weight spectrum. These data raise the question of whether OXT is a mediator of bone density, structure and fracture risk in women.  OXT pathways may provide novel targets for osteoporosis treatment.


Nothing to Disclose: MS, DAM, RLP, KTE, AK, KKM, EAL

25849 8.0000 FRI 356 A Oxytocin and Its Relationship to Bone Mineral Density and Hip Geometry Across the Weight Spectrum 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 349-367 7685 1:15:00 PM Osteoporosis - Risks and Treatments (posters) Poster

Sumit Majumdar*1, Robert G Josse2, Mu Lin3 and Dean Eurich3
1University of Alberta, Edmonton, AB, 2University of Toronto, Toronto, ON, Canada, 3University of Alberta


Background.  Type-2 diabetes and osteoporosis are both common, chronic, and increase with age while type-2 diabetes is also a risk factor for major osteoporotic fractures (MOF).  However, different treatments for type-2 diabetes affect fracture risk differently, with meta-analyses showing some agents increase risk (e.g., thiazolidinediones [TZD]) and some reduce risk (e.g., sitagliptin).  Our objective was to determine the independent association between new use of sitagliptin and MOF in a large population-based cohort study.

Methods.  In a new-user study design using a nationally representative US claims database of 72,738 insured patients with type-2 diabetes, we compared the incidence of MOF (hip, clinical spine, proximal humerus, distal radius) in new-users of sitagliptin vs non-users over a median 2.2 years followup.  We used 90-day time-varying sitagliptin exposure windows and controlled confounding by adjusting for clinical data, comorbidities, and time-updated propensity scores.

Results.  At baseline, the median age was 52 years, 54% were men, and median A1c was 7.5%.  There were 8894 new-users of sitagliptin and 63,834 non-users with a total of 181,139 person-years of followup.  There were 741 MOF, with 69 fractures (2.5 per 1000 person-years) among new-users of sitagliptin vs 672 fractures (4.4 per 1000 person-years) among non-users.  In multivariable analyses, sitagliptin was not associated with fracture (adjusted hazard ratio 1.1, 95%CI 0.8-1.5, p=0.5) although insulin (p<0.01), sulfonylureas (p<0.01), and TZDs (p=0.02) were each independently associated with an increased fracture risk.

Conclusions.  Even in a young population with type-2 diabetes, osteoporotic fractures were not uncommon.  New use of sitagliptin was not associated with fracture, but other commonly used second-line agents for type-2 diabetes were associated with increased risk.  These data should be considered when making treatment decisions for those with type-2 diabetes at particularly high risk of fractures.


Nothing to Disclose: SM, RGJ, ML, DE

27306 9.0000 FRI 357 A Does Sitagliptin Affect the Rate of Osteoporotic Fractures in Type-2 Diabetes? Population-Based Cohort Study 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 349-367 7685 1:15:00 PM Osteoporosis - Risks and Treatments (posters) Poster

Ernesto Maddaloni*1, Liane Tinsley2, Stephanie D'Eon2, Mary Larsen Bouxsein3, Nicola Napoli4, George L King5 and Hillary Keenan6
1Joslin Diabetes Center, Rome, Italy, 2Joslin Diabetes Center, 3Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, 4Universitá Campus Bio-Medico di Roma, Rome, Italy, 5Joslin Diabetes Center / Harvard Medical School, Boston, MA, 6Joslin Diabetes Center, Boston, MA


Increased bone fragility has been described in those with type 1 diabetes (T1D) with some studies suggesting an association with vascular complications. The Joslin 50-year Medalist Study (J50M) is a unique cohort of individuals with 50 or more years of T1D and a lower than expected prevalence of both micro- and macrovascular complications [1]. Congruently, we previously showed a low prevalence of fractures in this same group [2]. As bone density and  microarchitecture are a key determinants of bone strength, we evaluated factors associated with volumetric bone mineral density (vBMD), bone geometry and microarchitecture in J50M. High-resolution peripheral quantitative computed tomography (HR-pQCT) of the distal radius and tibia was performed in 20 J50M (7 males). The participants had a mean ± SD age of 65.4 ± 6.8 yrs, T1D duration of 56.3 ± 5.7 yrs, BMI of 25.9 ± 3.9 kg/m2, HbA1c of 7.1 ± 0.8%, eGFR of 78.8 ± 16.0ml/min/1.73m2, total cholesterol of 159.6 ± 36.1 mg/dL, triglycerides of 69.2 ± 17.1 mg/dL, HDL-C of 66.0 ± 22.8 mg/dL, LDL of 79.9 ± 20.0 mg/dL, and Vitamin D of 36.6 ± 15.4 ng/mL. Multivariable regression analyses adjusted for gender were performed using HR-pQCT measurements as dependent and clinical or biochemical markers as independent variables. Age was inversely associated with cortical vBMD at both the radius (β= -6.21, adjusted R2: 0.32, p=0.01) and tibia (β= -7.25 adjusted R2: 0.15, p=0.03). Unexpectedly, no relationship was found between age and trabecular vBMD at either the radius (p=0.20) or tibia (p=0.51). In contrast, tibia trabecular vBMD was inversely associated with LDL cholesterol levels (β= -0.60, p<0.05) and positively with BMI (β= 4.36, adjusted R2: 0.50, p=0.01). HbA1c was not significantly related to any measure of bone quality in either the trabecular or cortical regions. An inverse association between eGFR and trabecular vBMD was also found, but disappeared after correction for BMI, which was significantly and inversely related to eGFR. The unique characteristics of the J50M, having an average age of 65 years and T1D duration of 56 years, provide a unique opportunity to study the effects of long term T1D and the interaction of hyperglycemia and aging. Here we identified protection from age-related trabecular bone decline. Moreover, for the first time in aging T1D, we describe higher LDL cholesterol is associated with lower volumetric trabecular bone density. This is in accordance with evidence from in vitro and in vivostudies showing the deleterious effects of oxidized LDL cholesterol on osteoblast differentiation and on the crosstalk between osteoblasts and osteoclasts, supporting a role for dyslipidemia in the in the development of bone fragility.


Disclosure: GLK: Investigator, Sanofi, Investigator, Boehringer Ingelheim. Nothing to Disclose: EM, LT, SD, MLB, NN, HK

PP01-2 26707 10.0000 FRI 358 A Age and LDL Cholesterol Differentially Affect Bone Quality in Subjects with Extreme Duration Type 1 Diabetes 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 349-367 7685 1:15:00 PM Osteoporosis - Risks and Treatments (posters) Poster

Nina Le*1, Anne R Cappola2, Danielle Abraham3, Yi Huang3, Marc Hochberg3, Ram R Miller4, Michelle Shardell5, Gregory Hicks6, Denise Orwig3 and Jay Magaziner3
1Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, 2University of Pennsylvania, Philadelphia, PA, 3University of Maryland School of Medicine, Baltimore, MD, 4Novartis Institutes for Biomedical Medicine, Boston, MA, 5National Institute on Aging, Baltimore, MD, 6University of Delaware, Newark, DE


Background: The number of men included in studies of hip fracture consequences has been small. Therefore, relatively little is known about recovery from hip fracture in men and whether existing data from women apply to men. Growth hormone, testosterone and estrogen have anabolic effects on bone and low concentrations could affect recovery and increase risk of future fractures. We examined differences in anabolic hormones in men and women during the year after hip fracture.

Methods: The study included 157 men and 154 women aged ≥ 65 years enrolled in the Baltimore Hip Studies 7th cohort, a study that frequency matched (1:1) men and women on calendar time of hip fracture and hospital, and had blood assay data. Testosterone by LC/MS/MS, estradiol by LC/MS/MS, SHBG, and IGF-1 were assayed at four time points post hip fracture (within 15 days of admission, and at 2, 6, and 12 months) on fasting morning samples. Differences were evaluated between men and women at each timepoint using Wilcoxon rank-sum tests.

Results: Men had a mean age of 80 years and BMI of 25.6 kg/m2, and 90% were white. Women had a mean age of 81 years and BMI of 25.6 kg/m2, and 92% were white. Median testosterone levels in men were low after fracture at 146 ng/dL (IQR 70-265) and increased during recovery to 289 ng/dL (IQR 148-429) at 2 months, 332 ng/dL (IQR 233-430) at 6 months, and 344 ng/dL (233-450) at 12 months. Testosterone levels were low in women at all time points (median 10-12 ng/dL). Median estradiol levels were higher in men than in women, at 15 pg/mL (IQR 11-20), rising slightly during recovery to 19 pg/mL (IQR13-24) at 2 months, 22 pg/mL (IQR 16-29) at 6 months, and 23 pg/mL (IQR 18-30) at 12 months. Estradiol levels remained stably low in women, with median levels ranging from 3-5 pg/mL. SHBG levels were lowest immediately after fracture and increased by 2 months after recovery in both men and women, with higher SHBG levels in women. IGF-1 levels were similarly low at baseline in both men (86 ng/mL; IQR 69-124) and women (94 ng/mL; IQR 71-130) with improvement by 2 months to 115 ng/mL (IQR 84-147) in men and 116 ng/mL (IQR 90-152) in women.

Conclusions: In men who have sustained a hip fracture, testosterone, estradiol, and IGF-1 are lowest at the time of fracture and increase over the following year during recovery, while remaining low for age at all time points. Both absolute levels and recovery patterns of testosterone and estradiol differ between men and women, suggesting that hormonal interventions may provide a unique opportunity to assist recovery from hip fracture in older men.


Disclosure: RRM: Clinical Researcher, Novartis Pharmaceuticals, Clinical Researcher, Novartis Pharmaceuticals, Clinical Researcher, GlaxoSmithKline. DO: Consultant, Kinexum, Consultant, Sanofi, Consultant, Viking. JM: Consultant, Ammonett, Consultant, Orgranext, Consultant, Regeneron, Consultant, Novartis Pharmaceuticals, Principal Investigator, Eli Lilly & Company. Nothing to Disclose: NL, ARC, DA, YH, MH, MS, GH

25005 11.0000 FRI 359 A Testosterone, Estrogen, and IGF-1 in Men and Women Following Hip Fracture 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 349-367 7685 1:15:00 PM Osteoporosis - Risks and Treatments (posters) Poster

Jeffrey M Kroopnick*1, Anne R Cappola2, Yi Huang3, Marc Hochberg3, Ram R Miller4, Michelle Shardell5, Gregory Hicks6, Denise Orwig3 and Jay Magaziner3
1Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, 2University of Pennsylvania, Philadelphia, PA, 3University of Maryland School of Medicine, Baltimore, MD, 4Novartis Institutes for Biomedical Medicine, Boston, MA, 5National Institute on Aging, Baltimore, MD, 6University of Delaware, Newark, DE


Background: The number of men included in studies of the consequences of hip fracture has been small. Therefore, relatively little is known about recovery from hip fracture in men and whether existing data from women apply to men. Low concentrations of vitamin D could affect recovery and increase risk of future fractures. We examined differences in 25-hydroxyvitamin D (25OHD), calcium, and biomarkers of bone resorption and bone formation between men and women during the year after hip fracture.

Methods: The study included 157 men and 154 women aged ≥ 65 years enrolled in the Baltimore Hip Studies 7thcohort, a study that frequency matched (1:1) men and women on calendar time of hip fracture and hospital, and had blood assay data.  25OHD measured by LC/MS/MS, a biomarker of bone resorption (c-telopeptide; CTX), a biomarker of bone formation (procollagen type 1 amino-terminal propeptide; P1NP), calcium, and PTH were assayed at four time points post hip fracture (within 15 days of admission, and at 2, 6, and 12 months) on fasting morning samples. Differences were evaluated between men and women at each timepoint using Wilcoxon rank-sum tests.

Results: Men had a mean age of 80 years and BMI of 25.6 kg/m2, and 90% were white. Women had a mean age of 81 years and BMI of 25.6 kg/m2, and 92% were white. During the admission for hip fracture, median 25OHD levels were 15.2 ng/mL (IQR 10.5-20.5) in men compared with 23.7 ng/mL (IQR 16.3-29.7) in women (p < 0.001).  Levels were 21.1 ng/mL (IQR 13.1-26.8) vs. 29.4 ng/mL (IQR 20.0-34.8) at 2 months after the fracture, 21.0 ng/mL (IQR 14.3-28.9) vs. 30.5 ng/mL (IQR 23.1-37.6) at 6 months, and 23.9 ng/mL (IQR 14.7-27.9) vs 30.6 ng. mL (IQR 23.0-34.2) at 12 months (p< 0.001 for all comparisons).  Median serum calcium was statistically significantly lower in men upon admission (9.0 mg/dL vs 9.1 mg/dL, p=0.04) and at 2 months (9.5 mg/dL vs 9.7 mg/dL, p=0.02) post-hip fracture, while CTX was significantly higher in men on admission, and at 2 and 6 months after fracture (all p ≤ 0.01). Serum creatinine was slightly, but statistically significantly, higher in men at all measured time points. There were no significant differences between men and women in P1NP or PTH at any of the indicated time points.

Conclusion: 25OHD was significantly reduced in men at baseline and during the year after hip fracture, along with lower calcium levels and increased bone resorption compared to women. This underscores the importance of vitamin D assessment in older men, as well as missed opportunities in men for vitamin D supplementation and therapy to reduce bone resorption after hip fracture.


Disclosure: RRM: Clinical Researcher, Novartis Pharmaceuticals, Clinical Researcher, Novartis Pharmaceuticals, Clinical Researcher, GlaxoSmithKline. DO: Consultant, Kinexum, Consultant, Sanofi, Consultant, Viking. JM: Consultant, Ammonett, Consultant, Orgranext, Consultant, Regeneron, Consultant, Novartis Pharmaceuticals, Principal Investigator, Eli Lilly & Company. Nothing to Disclose: JMK, ARC, YH, MH, MS, GH

25006 12.0000 FRI 360 A Men Have Lower Vitamin D Levels after HIP Fracture and during Recovery Compared to Women 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 349-367 7685 1:15:00 PM Osteoporosis - Risks and Treatments (posters) Poster

Stephanie Kim*1, Bruce Ettinger2, Malini Chandra3 and Joan Chia-Mei Lo3
1Kaiser Permanente Oakland Medical Center, Oakland, CA, 2Kaiser Permanente Northern California, San Francisco, CA, 3Kaiser Permanente Northern California, Oakland, CA


BACKGROUND:  While Caucasian bone mineral density (BMD) reference data is recommended for defining osteoporosis among U.S. women regardless of ethnic origin, the use of ethnic T-scores has been shown to result in substantial reclassification in Asian women. This study examines the distribution of BMD and T-score classification among Northern California Chinese women in comparison with age-matched white women.

METHODS:  We used existing data from a population study of Kaiser Permanente Northern California (KPNC) women age 50-85 years old with BMD measured using a Hologic DXA scanner between 1997 and 2003 in whom femoral neck BMD and fracture risk were previously examined. Chinese women age 50-79 years old were identified and compared to an age-matched group of white women using a 1:5 ratio, with T-scores calculated at the femoral neck (FN), total hip (TH) and lumbar spine (LS) using Hologic (spine) or Hologic NHANES III (femoral neck and hip) reference data for white women (Caucasian T). An ethnic T-score was also calculated for Chinese women based on BMD reference data reported for Chinese Americans in New York (Walker MD et al, Osteoporos Int 2006; 17:878-887). The 10-year probability of hip fracture was estimated using FRAX with femoral neck BMD (version 3.1). 

RESULTS:  A total of 4039 Chinese women (44.8% age 50-59 years, 37.5% age 60-69 years, 17.7% age 70-79 years) and an age-matched cohort of 20,195 white women were identified. Compared to white women, BMD T-scores using the Hologic or Hologic NHANES III reference were significantly (p<0.001) lower at the FN, TH and LS when compared by race for each age group (median T-score 0.29-0.75 units lower for Chinese vs white women). However, ethnic T-scores calculated for Chinese women using Chinese American reference data resulted in an average change in T-score of +0.47, +0.36 and +0.48 units higher in the FN, TH and LS, respectively, with T-scores more comparable to those of age-matched white women. Use of ethnic T-scores reduced the overall percentage of Chinese women diagnosed with osteoporosis in the FN (16.7% to 6.6%), TH (9.8% to 3.2%), and LS (23.2% to 8.9%), with the largest percentage reduction in Chinese women aged 50-64 years. Of the subset of Chinese women aged 50-64 years diagnosed with FN osteoporosis by Caucasian T-score but FN osteopenia by ethnic T-score, less than1% had a FRAX-hip score of 3% or higher.

CONCLUSION:  The use of Chinese American BMD reference data for U.S. Chinese women yields higher (ethnic) T-scores by about 0.4-0.5 units. As a result, a large proportion of postmenopausal Chinese women had DXA results that were reclassified from osteoporosis to osteopenia. The reduction in excess osteoporosis labeling when ethnic T-scores are used may be particularly relevant for younger Chinese women who are at lower fracture risk.


Disclosure: JCML: Principal Investigator, Sanofi, Principal Investigator, Astra Zeneca, Principal Investigator, Sanofi. Nothing to Disclose: SK, BE, MC

25461 13.0000 FRI 361 A Using Ethnic-Specific BMD Norms in Chinese American Women 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 349-367 7685 1:15:00 PM Osteoporosis - Risks and Treatments (posters) Poster

Cemre Robinson*1, Nataliya Buxbaum2, Kristin Baird3, Amit Tirosh4, Maya Beth Lodish4 and Steven Z. Pavletic2
1The Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, 2Experimental Transplantation and Immunology Branch, National Cancer Institute Center for Cancer Research, National Institutes of Health, 3Division of Clinical Evaluation, Pharmacology and Toxicology, Office of Cellular, Tissue and Gene Therapies, Center for Biologics Evaluation and Research, Food and Drug Administration, 4National Institutes of Health, Bethesda, MD


Background: Chronic graft versus host disease (cGVHD) is a major complication after hematopoietic stem cell transplantation (HSCT).   In survivors of childhood cancer, diminished bone mineral density (BMD) has been associated with endocrine dysfunction, total body irradiation (TBI), and prolonged use of glucocorticoids; however BMD in children with cGVHD has not been well-characterized.

Methods:  Data from 28 patients with cGVHD seen at the NIH CC between the years 2005-2015 were analyzed [18 M, mean age 13.0±4.8].  BMD in AP spine, total hip, femoral neck, and 1/3 forearm, was measured by dual energy x-ray absorptiometry (DEXA).  Age- and gender-adjusted z-scores were calculated using the Bone Mineral Density in Childhood Study calculator.  Clinical and anthropometric data were obtained at the time of DEXA examination. We studied the relationship between BMD, duration of cGVHD, TBI, intensity of immunosuppression, number of systemic therapies, and sum of NIH cGVHD organ scores.  Paired Student's t-tests were used to compare DEXA measurements in different sites, independent t-tests to analyze continuous variables, and chi-squared tests for categorical variables.  Pearson product was used for analysis of correlation between variables.  p-value set <0.05.

Results: The mean BMD z-scores for each individual site were as follows:  -1.97±1.90 at spine,-3.35±3.41 at femoral neck, -2.96±2.26 at total hip, and -2.87±2.64 at 1/3 forearm.  The femoral neck z-score was lower than the spine (p=0.015), suggesting that cortical bone may be more severely affected than trabecular bone.  The majority of patients (78.6%) had z-scores consistent with impaired BMD, as defined by a z-score ≤-2.0.  Patients with impaired BMD were more likely to have received TBI conditioning and higher intensity of immunosuppression, (p=0.033 and 0.04, respectively).  The z-score at the 1/3 forearm had positive correlation with BMI (rp=0.4, p=0.03), and with age at diagnosis (rp=0.4, p=0.02).  In addition, the intensity of immunosuppression correlated negatively with spine z-score (rp=-0.5, p=0.015) and with 1/3 forearm z-score score (rp=0.4, p=0.04).  There were no statistically significant differences between patients with impaired BMD and normal BMD in regards to the duration of cGVHD, number of systematic therapies, or sum of NIH organ score. 

Conclusions:  Our data shows that pediatric BMT survivors with cGVHD have severely impaired BMD.  In our patient cohort, we describe site-specific bone loss with worse values at the femoral neck.  Our results suggest that these patients may be at greater risk for impaired BMD following TBI conditioning and higher intensity of immunosuppression.  Recognition of diminished bone mineral density is important in cancer survivors with cGVHD.  Early intervention to minimize risk factors and optimize bone health in pediatric patients is critical as the greatest bone accrual occurs during adolescence.


Nothing to Disclose: CR, NB, KB, AT, MBL, SZP

25902 14.0000 FRI 362 A Impaired Bone Mineral Density in Pediatric Chronic Graft-Versus-Host-Disease 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 349-367 7685 1:15:00 PM Osteoporosis - Risks and Treatments (posters) Poster

Asma Al-Zougbi*1 and Amal Shibli-Rahhal2
1University of Iowa Hospitals and Clinics, Iowa City, IA, 2University of Iowa, Iowa City, IA


Background: Patients with Duchenne Muscular Dystrophy (DMD) exhibit low bone mineral density (BMD) and increased fracture risk, which is often detrimental to their mobility. Since many of these patients receive corticosteroids to delay muscle fiber loss, immobility and death, they also exhibit delayed puberty and short stature, so it important to account for delayed skeletal development when interpreting their BMD.  A recent study showed that while BMD adjustment for bone size in DMD patients did show bone mineral deficit, this deficit was not commensurate with that seen when BMD was adjusted to chronological age according to standard DXA protocols.  Another method of BMD adjustment is to use bone age rather than chronological age, since bone age is expected to be delayed in these patients. However, to our knowledge, there is no published data on BMD adjustment to bone age in this patient population.  

Methods: We report the cases of 5 patients with DMD who were seen in our Metabolic Bone Disease Clinic. DXA scans and wrist x-rays to estimate bone age were obtained as part of their routine clinical care. Z-scores were initially determined based on their chronological age. We then assigned to each patient a “corrected” birth date that was calculated based on bone age and we determined bone-age-corrected Z scores.

Results: Patient A was diagnosed with DMD at the age of 3 years, and has been on prednisone since age 7. At the age of 19 years and 1 month, his bone age was 17 years.  His Z-score at the left radius was -6.0 based on chronological age and -5.3 based on bone age.

Patient B was diagnosed with DMD at the age of 7 months and has been on prednisone since the age of 6 years. At the age of 16 years and 2 months, his bone age was 13 years. His Z-score at the right femoral neck was -4.3 based on chronological age and -4.1 based on bone age.

Patient C was diagnosed with DMD at the age of 5 years and has been on deflazocort since the age of 6. At the age of 17 years and 5 months, his bone age was 12 years and 6 months. His Z-score at the lumbar spine was -1.3 based on chronological age and +1.3 based on bone age.

Patient D was diagnosed with DMD at the age of 3 years and has been on deflazocort since the age of 6. At the age of 17 years and 11 months, his bone age was 13 years. His Z-score at the lumbar spine was -3.3 based on chronological age and -1 based on bone age.

Patient E was diagnosed with DMD at the age of 7 years, and was treated with deflazocort between the ages of 8 and 12 years, followed by prednisone since then. At the age of 22 years and 8 months, his bone age was 18 years.  His Z-score for lumbar spine was -2.3 based on chronological age and -1.6 based on bone age.

Conclusion:  The use of bone age rather than chronological age for adjustment of BMD for in DMD patients on corticosteroids might better reflect their bone mineral status. Additional research is needed to better determine the trajectory of skeletal development and its effect on BMD in this patient group.


Nothing to Disclose: AA, AS

26743 16.0000 FRI 364 A Adjustment of Bone Density for Chronological Age in Children with Duchenne Muscular Dystrophy Overestimates Their Bone Mineral Deficit 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 349-367 7685 1:15:00 PM Osteoporosis - Risks and Treatments (posters) Poster

Maria Marta Sarquis Soares*1, Michelle S Diniz2, Barbara C Silva3, Marcio W Lauria4, Jackson P Machado5, Adriana Maria Kakehasi6 and Antonio L Teixeira-Jr7
1Universidade Federal de Minas Gerais, Belo Horizonte MG, Brazil, 2Santa Casa de Belo Horizonte, Belo Horizonte, Brazil, 3Santa Casa de Belo Horizonte, 4FM/ Univ Fed de Minas Gerais, Belo Horizonte MG, Brazil, 5Santa Casa de Belo Horizonte, Brazil, 6Universidade Federal de Minas Gerais, Belo Horizonte, Brazil, 7Federal University of Minas Gerais


Introduction: Psoriasis is a chronic inflammatory disease characterized by increased expression of several cytokines. Interferon (IFN)- γ, tumor necrosis factor (TNF)- α and interleukin (IL)-6key cytokines that contribute to the initiation and perpetuation of psoriatic lesions, are also known to play a role in the pathogenesis of osteoporosisBased on this observation, one could hypothesize that psoriatic patients may be more susceptible to osteoporosis than healthy individuals.


Objective: To determine serum levels of pro-inflammatory cytokines and bone turnover markers (BTM) in patients with psoriasis and healthy controls, and to correlate these measurements with bone mineral density (BMD).


Methods: We studied 34 patients with psoriasis and 35 age- and sex-matched healthy controls. We assessed BMD by dual energy x-ray absorptiometry (DXA), serum BTMs (OPG, CTX-I and RANKL) and cytokines (IL-2, IL-4, IL-6, IL-10, IL-12, IL-17A, IL-23, INF -γ, TNF-α) in all participants. Statistical analyzes were done using SPSS 15.0 and a p value <0.05 was considered significant. The study was approved by the local Ethical Committee.


Results: Psoriatic and healthy subjects were well matched in age (mean age (±SD) of 46.2 ± 14.6 vs. 40.6 ± 12.1 years, respectively. The frequency of men was also similar between the groups (61.8% in the psoriatic group vs. 54.3% in the control group. The average duration of the disease in individuals with psoriasis was 12.5 years. Patients with psoriasis had statistically higher levels of OPG, IL-12, TNF-α, IL-10, IL-6, IL-4 and IL-2 than controls. Patients with psoriasis under systemic treatment had higher levels of TNF-α and CTX-I than psoriatic patients under topical treatment onlySerum levels of OPG, IL-6, IL-17A and IL-23 were inversely correlated with BMD in the entire group.


Conclusion: We have shown, in agreement with previous studies, that several cytokines are increased in psoriatic patients as compared to healthy controlsand that they are inversely correlated with BMD. In addition, the serum level of OPG, a BTM, is increased in subjects with psoriasis, and the serum concentration of the bone resorption marker CTX-I is higher in patients with a more severe form of the diseasesuggesting that the systemic inflammation observed in subjects with psoriasis may influence other systems such as bone metabolism. 


Nothing to Disclose: MMSS, MSD, BCS, MWL, JPM, AMK, ALT

26515 17.0000 FRI 365 A Levels of Pro-Inflammatory Cytokines and Proteins of the Osteoprotegerin-RANKL System and Its Correlation with Bone Mineral Density in Patients with Psoriasis and Controls 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 349-367 7685 1:15:00 PM Osteoporosis - Risks and Treatments (posters) Poster

Ma. Conchitina Manas Fojas*1, Steven Wai Ing2, Laura Phieffer1, Julie Stephens1 and Lauren Southerland1
1The Ohio State University Wexner Medical Center, Columbus, OH, 2The Ohio State University, Columbus, OH


Introduction: Professional societies advocate deployment of a Fracture Liaison Service (FLS) to prevent secondary fractures, but care gaps persist. International Osteoporosis Foundation provided a framework to evaluate FLS programs and National Quality Forum endorsed a set of core measures for adoption by The Joint Commission. We evaluated the effectiveness of the fracture prevention program (FPP) at the Ohio State University Wexner Medical Center (OSU) to meet core measures.

Hypothesis:  Core measures were better achieved through coordinated efforts between Endocrinology, Orthopedics, and Emergency Medicine.

Methodology: We reviewed charts of 452 patients admitted to OSU due to fragility fracture (June 2010-October 2015). Of these, 318 were placed in three groups based on time of admission: FLS (2010-2012), FPP (2014-2015), or Control (2012-2014, without FLS or FPP). FLS was led by an orthopedic nurse practitioner (NP). FPP consisted of fragility fracture identification by Emergency Medicine or Orthopedics, inpatient Endocrinology consultation, and referral to outpatient High-Risk Osteoporosis Clinic. Primary outcomes were: completion of all 5 core laboratory tests (CBC, renal function, liver function, calcium, and 25-hydroxy Vitamin D), DXA scan scheduled before discharge, DXA scan completed by 3 months post discharge, osteoporosis drug continued or started during hospitalization, and recommendation for outpatient follow up for osteoporosis. The proportion of patients with each of the outcomes and group comparisons were done using Chi-square or Fisher’s Exact test.

Results: The 318 subjects consisted of 122 FLSs, 110 FPPs, and 86 controls. Most were female (74%), Caucasian (88%), and the mean age was 71 years.  53% were admitted into Orthopedics service and 81% had surgery. All 5 labs were completed in 83% of FPPs, only 36% in FLS and controls (p<0.001). Over half (56%) FPP were scheduled for DXA prior to discharge, while only 2% controls and 0% FLS. A DXA scan within 3 months was completed in 55% FLS, 35% FPP, and 7% control (p<0.001). 2-10% continued or started osteoporosis drug during hospitalization which was not statistically different (p=0.073). Nearly all FLS had recommendation for follow up (98%) versus 77% FPP and 13% controls (p<0.001).

Discussion: Compliance in completing all lab tests, highest in FPP may be related to inpatient endocrinology consultation.  DXA scan completion within 3 months and scheduling follow up visits were highest in FLS, perhaps due performance of “patient navigator” functions by the NP post discharge. Recommendation for drug was similar, but this does not imply equal initiation and adherence, since drugs were typically prescribed in the outpatient setting.

Conclusion: This study highlighted the effectiveness of FLS and FPP in adherence to core measures and demonstrated the need for seamless integration between inpatient and outpatient settings.


Nothing to Disclose: MCMF, SWI, LP, JS, LS

27654 18.0000 FRI 366 A Evolution of a Fracture Prevention Program : A Review of Our Experience at the Ohio State University 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 349-367 7685 1:15:00 PM Osteoporosis - Risks and Treatments (posters) Poster

Cristina Capatina, Mihai Berteanu and Catalina Poiana*
Carol Davila University of Medicine and Pharmacy, Bucharest, Romania


  Introduction.   Postmenopausal osteoporosis is associated with an increased risk of fragility fractures and recurrent falling. The potential impact on the quality of life is insufficiently characterised.

  Objective. To analyse the quality of life in postmenopausal women with osteoporosis (compared to an age-matched group of healthy women) and possible correlations with the numbers of prevalent fragility fractures and falls.

  Material and methods. The study group consisted of 72 women with postmenopausal osteoporosis (defined by central densitometry). The control group consisted of 48 age-matched women with normal bone mass and no positive history for fragility fractures. We assessed the quality of life using the osteoporosis assessment questionnaire (OPAQ) and the EuroQoL. The serum concentration of 25OHD was measured by electrochemiluminiscence in all subjects.

  Results. VD deficiency was markedly prevalent in both groups. Fractures were present only in the osteoporotic group: 35 cases had at least one prevalent fragility fracture.

The patients with osteoporosis had significantly worse scores on bodyimage and pain (OPAQ) (p=0.006 and 0.029 respectively), borderline significance for lower visual assessment (VAS) score (EuroQoL) (p=0.049). Also the number of fallers (subjects who had at least one fall episode in the previous year) was significantly higher in the study group compared to the control group.(17 versus 4, p=0.031)

In the osteoporotic group, the fearfall and bodyimage were significantly worse in patients with falling episodes in the previous year (p =0.022 and 0.041, respectively) and correlated with the number of falls. As expected, the number of fractures was also significantly correlated with the number of falls (p=0.000). However, if corrected for the falls number, the number of prevalent fractures is not significantly correlated to any of the QoL items.

  Conclusions. Women with postmenopausal osteoporosis have impaired quality of life compared to age-matched postmenopausal women.  Affected quality of life subscores are significantly correlated with the number of falls in the previous year.


Nothing to Disclose: CC, MB, CP

27489 19.0000 FRI 367 A The Quality of Life in Women with Postmenopausal Osteoporosis 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Bone, Calciotropic Hormones & Vitamin D Friday, April 1st 3:15:00 PM FRI 349-367 7685 1:15:00 PM Osteoporosis - Risks and Treatments (posters) Poster

Carolina Panico1, Resmi Premji*2, Sonia Q Doi3 and Eric S Nylen4
1Veterans Affairs Medical Center, Washington, DC, 2George Washington University, Washington, DC, 3Uniformed Svcs Univ Hlth Sci, Bethesda, MD, 4VA Medical Center, Washington, D.C.


Background: Exercise can improve renal function in diabetic subjects with overt nephropathy, however it is still unclear whether exercise in chronic kidney disease (CKD) with estimated glomerular filtration rate (eGFR) above 60 mL/min/1.73 m2 (i.e., CKD 1- 2) could delay the progression of and/or reverse kidney damage. Furthermore, the clinical diagnosis of CKD 1 and 2, based on eGFR measurements may not be accurate, especially in the elderly, and this may explain the lack of information regarding the effect of exercise at early stages of CKD. Detection of urinary protein levels in exosomes is a putative noninvasive method to monitor the progression of diabetic kidney disease. In this preliminary study, we measured podocyte proteins Wilms Tumor -1 (WT-1) and nephrin from urinary exosomes, and exosomal marker CD 9 before and after a bout of exercise in elderly diabetic subjects with microalbumiuria and early CKD.

Methods: Our study subjects consisted of elderly individuals with type 2 diabetes mellitus, microalbuminuria, and eGFR between 60 and 90 mL/min/1.73 m2 and non-diabetic controls. All subjects exercised for 30 minutes and urine was collected before and immediately after the exercise session. We isolated urinary exosomes using differential centrifugation. Subsequently exosomal proteins were resolved by SDS-PAGE, and Western Blot was performed for the target proteins WT-1 and nephrin. We also measured albumin/creatinine ratio before and after the exercise session.

Results: We observed that baseline WT-1 and nephrin were elevated in urinary exosomes of diabetic subjects compared to non-diabetic individuals. In the urine collected immediately after an acute session of exercise, both WT-1 and nephrin were markedly and consistently decreased compared to the pre-exercise value. Similar findings were observed for the exosomal marker protein CD9.  On the other hand, levels of albuminuria varied after exercise, and the changes were not consistent among the individuals.

Conclusion: The urinary exosomal podocyte markers WT-1 and nephrin were elevated prior to exercise. There was an acute loss of these proteins as well as a decrease in exosomes following exercise, a pattern not observed with albumin. Thus, both WT-1 and nephrin were shown to be responsive to exercise while the response of albumin was variable. Future studies will aim to explore the durability of these exosomal changes and its relationship to diabetic kidney damage and response to long-term exercise training.


Nothing to Disclose: CP, RP, SQD, ESN

26552 1.0000 FRI 651 A Exosomal Response to Exercise in Diabetic Kidney Disease 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Sivaporn Sivasinprasasn*, Pongpan Tanajak, Wanpitak Pongkan, Wasana Pratchayasakul, Siriporn C Chattipakorn and Nipon Chattipakorn
Chiang Mai University, Chiang Mai, Thailand


Estrogen deprivation has been shown to result in more severe cardiac tissue damage and dysfunction after myocardial ischemia and reperfusion (I/R).  In estrogen-deprived condition, chronic high-fat diet consumption accelerated and increased the severity of cardiometabolic dysfunction.  Both estrogen supplement and DPP-4 inhibitor; vildagliptin, has been reported to reduce myocardial damage due to cardiac I/R injury.  However, their effects on the heart in obese-insulin resistant and estrogen deprived condition remain unknown.  Bilateral ovariectomized (O) rats (n= 36) were divided to receive either normal diet (NDO) or high-fat diet (HFO) for 12 weeks, followed by being treated with vehicle, estrogen (E; 50 µg/kg) or vildagliptin (Vil; 3 mg/kg) for 4 weeks.  Sham-operated female rats fed with normal diet (NDS) were used as a control.  After 4-week treatment, heart rate variability (HRV), echocardiography and metabolic parameters were examined.  Then, 30-min ischemia followed by 120-min reperfusion was performed, and the infarct size and cardiac mitochondrial function was determined.  After being ovariectomized for 12 weeks, both NDO and HFO rats exhibited obese-insulin resistant condition.  Treatments with E and Vil effectively reduced fasting plasma levels of glucose, insulin and HOMA index in both NDO and HFO rats (P<0.05, n=6/group).  Both treatments also improved cardiac function via restoring HRV and %EF (P<0.01, when compared with vehicle group in both NFO and HFO rats).  After I/R, cardiac mitochondrial function was preserved by both E and Vil, as indicated by decreased ROS level, mitochondrial depolarization and swelling in ischemic myocardium, when compared with vehicle-treated NDO and HFO groups (P<0.05, n=6/group).  Moreover, the level of infarct size in both NDO and HFO rats was significantly decreased by both E (P=0.01, n=6/group) and Vil (P=0.03, n=6/group) in consistent with the reduced levels of oxidative stress (P<0.05, n=6/group) as well as the apoptotic protein expression (P<0.05, n=6/group) in the ischemic myocardium.  These findings clearly demonstrated that treatments with estrogen and vildagliptin effectively improved metabolic status, and share similar efficacy in reducing myocardial infarction and protecting cardiac mitochondrial function against I/R injury in estrogen-deprived obese-insulin resistant rats.


Nothing to Disclose: SS, PT, WP, WP, SCC, NC

25257 2.0000 FRI 652 A DPP-4 Inhibitor and Estrogen Attenuate Metabolic Dysfunction and Mitochondrial Impairment from Cardiac Ischemic-Reperfusion Injury in Obese-Insulin Resistant and Estrogen-Deprived Rats 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Nattayaporn Apaijai*1, Narattaphol Charoenphandhu2, Jitjiroj Ittichaichareon1, Panan Suntornsaratoon2, Nateetip Krishnamra2, Ratchaneevan Aeimlapa2, Siriporn C Chattipakorn1 and Nipon Chattipakorn1
1Chiang Mai University, Chiang Mai, Thailand, 2Mahidol University, Bangkok, Thailand


Both type 2 diabetes mellitus (T2DM) and an estrogen deprivation have been shown to be associated with the development of cardiovascular disease.  In T2DM, adverse left-ventricular (LV) remodeling together with cardiomyocyte growth, inflammation, fibrosis, and arrhythmia have been observed.  Estrogen deprivation has been found to be associated with coronary heart disease.  However, the role of estrogen deprivation on adverse LV remodeling in T2DM has not been clearly elucidated.  We hypothesized that estrogen-deprivation aggravates adverse LV remodeling in T2DM rats.  Wild-type (WT) and Goto-Kakizaki (GK) rats (9 months old, n=12/group) were used in this study.  Rats in each group were divided into 2 subgroups to receive either sham (S) operation or bilateral ovariectomy (O) (n=6/subgroup).  Four months after operation, rats were sacrificed, and the heart was rapidly excised for the determination of metabolic parameters and adverse LV remodeling.  The level of connexin-43 phosphorylation (p-Cx43) was determined for the function of gap junctions.  Cardiac fibrosis was determined using picrosirius red staining and TGF-β expression (a fibrosis inducer).  LV hypertrophy was determined using the Heart weight/Body weight (HW/BW) ratio, H&E staining, and NFκB (an inflammatory marker) expression.  Our results showed that all GK rats, a genetically-engineered T2DM rats, had hyperglycemia with hypoinsulinemia.  In ovariectomized group, body weight was increased in both WTO and GKO rats, compared with their sham-operated rats in the same genotype (p<0.05).  However, GKO rats had lower plasma glucose than GKS rats.  In the cardiac tissues, GKS and GKO rats had lower p-Cx43, compared with WTS rats (p<0.05), and there was no statistical significant between GKS and GKO rats, indicating that gap junction function was greater impaired in GK rats, and that estrogen deprivation had no influence on p-Cx43 in GK rats.  Picrosirius red staining and TGF-β expression were increased in both GKS and GKO rats, compared with WTS rat (p<0.05), and no significant difference was found between GKS and GKO rats, indicating that cardiac fibrosis was developed in GK rats.  Furthermore, HW/BW and cardiomyocyte cross-sectional area were increased in GKS and GKO rats, compared with WTS rats.  However, GKO rats had a higher HW/BW ratio and cardiomyocyte cross sectional area than GKS rats and WTO rats (p<0.05).  Our data indicated that LV hypertrophy was observed in GK rats, and that estrogen deprivation aggravated LV hypertrophy in these T2DM rats.  Moreover, NFκB expression was increased in GKS and GKO rats, compared with WTS rats, and NFκB expression was higher in GKO rats, compared with GKS and WTO rats (p<0.05).  All of these findings indicated that although an estrogen deprivation reduced blood glucose level in T2DM rats, it aggravated adverse LV remodeling by provoking cardiac inflammation in these genetically-engineered T2DM rats.


Nothing to Disclose: NA, NC, JI, PS, NK, RA, SCC, NC

25241 3.0000 FRI 653 A Estrogen Deprivation Aggravates Adverse Left Ventricular Remodeling in Type 2 Diabetic Rats 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Alison Heather*1, Obialunanma Vivian Ebenebe2 and Lucinda Sue McRobb3
1University of Otago, Dunedin, New Zealand, 2University of Otago, 3Macquarie University, Sydney, Australia


Calcification is a hallmark of advanced atherosclerosis and is associated with increased risk of myocardial infarction and stroke. Vascular calcification is an active and complex process that involves numerous mechanisms responsible for calcium depositions in arterial walls. Estrogens used for hormone replacement therapies may interfere with processes occurring in the vessel wall. To study whether estradiol treatment affects calcification of advanced atherosclerotic lesions, we implanted silastic estradiol (E2) capsules into 34-week old atherosclerosis-prone mice (apolipoprotein E-null mice). After 8 weeks, the atherosclerotic calcified lesion area was measured in the aortic sinus. There was no significant change in total lesion area with E2 treatment; however, there was an increase in calcified area relative to total area (0.5801 vs 1.894, p<0.001). E2 mediates its effects via estrogen receptor (ER)-α or –β so we measured ERα and –β levels by immunohistochemistry. E2 had no effect on ERα levels but did decrease ERβ (81±1% vs 11±5% positive nuclei, p<0.0001). To begin to understand the molecular underpinnings that alter vascular calcification processes, we exposed vascular smooth muscle cells (VSMC) to 4nM E2. VSMCs exposed to E2 showed increased calcification, as measured by von Kossa staining and calcium assay (by 175±7.9%, p<0.001). The in vivo data suggested that calcification associates with decreased ERβ so we next treated cells with E2 in the presence of ERβ antagonist (PHTPP), and found that the antagonist increased calcification (by 231±6.6 %, p<0.01). To explore further the mechanism of VSMC differentiation to a mineralizing cell induced by E2, we used digital PCR to measure expression of osteoblast cell markers. E2 treatment significantly decreased expression of 2 inhibitors of calcification, matrix Gla protein (MGP, 209 vs 148 copies/μL, p<0.05) and osteopontin (OPN, 149 vs 109 copies/μL p<0.05). By contrast, E2 increased calcification and/or osteoblast markers, osteocalcin (107 vs 150 copies/μL p<0.05), bone sialoprotein (BSP, 170 vs 202 copies/μL p<0.05), and collagen I (200 vs 224 copies/μL p<0.05) and II (381 vs 407 copies/μL p<0.05). To test whether VSMC express these same osteoblast markers in vivo, laser capture microdissection of VSMCs from non-calcified areas immediately adjacent to calcified areas was performed. Digital PCR showed MGP was decreased (17.9 vs 9.3 copies/μL p<0.05) while BSP (11.5 vs 32.4 copies/μL p<0.05) and collagen I (41.1 vs 72.1 copies/μL p<0.05) were increased. From these findings, we now speculate that E2 can drive calcification in advanced, non-growing, atherosclerotic lesions by promoting the dedifferentiation of VSMC to osteoblast-like cells. These findings require some consideration as E2 therapies are often prescribed for older females that may have advanced atheroma.


Nothing to Disclose: AH, OVE, LSM

27219 7.0000 FRI 657 A Estradiol Promotes Atherosclerotic Lesion Calcification in Aged, Female Mice 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Harriette Rosen Mogul1, Ruth G Freeman*2, Khoa Nguyen3 and Philipp E Scherer4
1New York Med Coll, Valhalla, NY, 2Montefiore Med Ctr, New Rochelle, NY, 3New York Medical College, Valhalla, NY, 4University of Texas Southwestern Medical Center, Dallas, TX


INTRODUCTION Android obesity is a major determinant of insulin resistance, metabolic syndrome (MS), and cardiovascular disease, whereas gynoid adiposity lowers these risks. Android to gynoid fat ratio (A/G ratio), an important new index of adiposity, is easily measured by DXA, providing precise, accurate assessment of these critical regions of interest.(1)  A/G ratio is the best correlate of markers of MS in cross-sectional studies of children,(2) and young adults,(3) and the strongest independent predictor of MS,(4) incident myocardial infarction,(5) and mortality in women(6) in longitudinal analyses. A/G ratio has not been evaluated in clinical trials of high risk females. 

METHODS EMPOWIR was a double-blind, placebo controlled, 12-month (12m) trial of women with ≥20lb midlife weight gain, normal glucose tolerance, and increased Area-under-the-curve insulin.(7) Subjects attended 4 nutrition workshops to introduce the carbohydrate modified diet (CMD), (45-50% carbohydrates, 30-35% protein, and 15-20% fat), a food exchange program, promoting increased intake of vegetables, low glycemic index fruits, low-fat protein & dairy products, elimination of free sugars, and notable restriction of 3 additional carbohydrates (starches) to after 4PM.  Participants were then randomized to CMD with metformin (M) or with metformin + rosiglitazone (MR) for 12 months, or to CMD alone for 6 months, with re-randomization to active treatment for months 7-12. Baseline and 12m body composition were measured by GE Lunar Prodigy DXA and reviewed by a single observer, with A/G ratio as previously defined.(1,4,8-10). Total adiponectin was measured with a commercial ELISA kit (Millipore/Linco) in the laboratory of Dr. Philipp Scherer. Paired t-tests were used to compare mean A/G ratios at baseline and 12m in 27 of 32 study completers (46.7± 6.5 years, BMI 30.8±2.8kg/m2, 50% white) with available data. Multiple regression was used to assess the relationship of 12-month A/G ratio with %-change in total adiponectin (Adp), race, baseline BMI, age, and initial treatment assignment (SPSS 22). 

RESULTS. Mean 12 month A/G ratio declined significantly in D and M arms (.986±.075 to .923±.098 (P=.011) and .961±.062 to .914±.091 (P=.018), and .974±.006 to .904±.150 (P= 0.007) in the total group. % change in Adp was the single best predictor of 12-month A/G ratio, Ɓ = - 0.001, R2 = .190, P=0.033, with no significant change after adjustment for included covariates. 

CONCLUSIONS We believe this is the first demonstration of reduction in A/G ratio in high risk women and suggest the EMPOWIR interventions merit additional investigation in larger scale long term studies.


Disclosure: PES: Investigator, Takeda, Investigator, Merck & Co.. Nothing to Disclose: HRM, RGF, KN

24449 9.0000 FRI 659 A Carbohydrate Modified Diet and Insulin Sensitizers Reduced Android to Gynoid Fat Ratio at 12 Months in EMPOWIR (Enhance the Metabolic Profile of Women with Insulin Resistance): A Double Blind, Placebo Controlled Randomized Trial of Normoglycemic Women with Midlife Weight Gain (NCT00618072) 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Jonathan Eli Arbelle*1, Morris Mosseri2 and Avi Porath1
1Maccabi Health Services, Tel Aviv, Israel, 2Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel


The role of HMG-COA inhibitors (statins) in reducing serum LDL-C levels and risk ratio for cardiovascular disease (CVD) in primary prevention is well established. Statin therapy is generally considered safe, with a low rate of reversible side effects.  Recently, statin therapy has been found to be associated with an excess risk for incident diabetes mellitus (DM). This hyperglycemic effect might induce diabetic complications and offset its cardiovascular protective effect. Whereas the consensus is that the benefits of statin therapy outweigh the risks in appropriately selected patients for secondary CVD prevention, the risk-benefit balance of statin treatment in primary prevention is less clear. The aim of this study was to assess the risk-benefit of statin therapy in a large, unselected population without DM or CVD. In a retrospective, large, population-based study we assessed the effect of statin therapy on the incidence of DM and coronary heart disease (CHD) among 351,917 subjects aged 40-70 years old without DM or CHD at baseline, members of Maccabi Health Services, Israel’s second largest health service provider. From 2010-2014, 6,359 (1.9%) new cases of CHD and 14,221 (4.0%) of DM were recorded. Observed outcomes were evaluated according to dose-intensity regimen, level of adherence to statin therapy and retrospectively determined baseline 10-year cardiovascular (CV) mortality risk (European SCORE). Statin therapy was used by 15.8% of the study population. 89.6% of statins users were prescribed low-intensity dose regimens. Statin therapy of low-intensity and above 50% adherence increased DM incidence in patients at low or intermediate baseline CV risk, but not in patients at high CV risk. At all intensity and adherence levels, statin therapy reduced CHD incidence in patients at intermediate or high baseline CV risk, but not in low-risk patients where only high intensity statins reduced CV risk. Expressed as numbers needed to treat (NNT) or harm (NNH), in patients at low CV risk practically no CV protective benefit was obtained and the NNH (incident DM) for low-intensity dose regimens and above 50% adherence was 40; in patients at intermediate CV risk the NNT was 125 with an NNH of 50; in high CV risk patients the NNT was 29 with an NNH of 200. Assessing the CV risk of patients can determine the balance between benefit and harm to patients with statin therapy. Primary prevention of CVD using statins should take into consideration the risk of statin-induced DM. Our findings support the current guidelines that call for individualised treatment decisions in healthy at-risk subjects.


Nothing to Disclose: JEA, MM, AP

24540 10.0000 FRI 660 A Primary Cardiovascular Disease Prevention with Statin Therapy: The Risk of Diabetes Mellitus and Cardiovascular Benefit per Euro-Score Risk 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Gabriela Heiden Telo*1, Rita Fernandes2, Felipe Vogt Cureau1, Laura Augusta Barufaldi2, Luciane Gaspar Guedes2, Cristina Kuschnir2, Beatriz D'Agord Schaan3 and Katia Vergetti Bloch2
1Universidade Federal do Rio Grande do Sul, 2Universidade Federal do Rio de Janeiro, 3Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil


Introduction:Considering that the prevalence of obesity among adolescents is increasing worldwide in recent decades, development of prevention strategies for high-risk groups is likely to have the largest impact in reducing the occurrence of diabetes and its chronic complications. Hemoglobin A1c (HbA1c) has been used as a primary measure to identify patients at high-risk for diabetes and cardiovascular diseases.

Aim: To evaluate the prevalence of high-risk HbA1c levels based on demographics and clinical characteristics among students from 12 to 17 years of age who participated in the Study of Cardiovascular Risk in Adolescents (ERICA).

Methods:We developed a cross-sectional study to evaluate a national school-based sample of adolescents in all capitals of the five Brazilian macro-regions, as well as their municipalities with more than 100,000 people. Blood sample were collected in schools and then analyzed in a single laboratorial center. Data analysis was adjusted for sampling design. HbA1c ≥5.7% was used to determine participants at high-risk for diabetes. Analyses were based on means and 95% confidence intervals (CI). Prevalence of high-risk HbA1c levels was evaluated by age, sex, macro-region, type of school (public or private), race/ethnicity, and weight status (age and sex specific z-score body mass index [z-BMI]).

Results:The mean HbA1c level among the total sample of 37,804 adolescents was 5.38% (95%CI 5.37 to 5.39). Overall, 20.5% (95%CI 19.1 to 22.0) had HbA1c values ≥5.7%; 23.6% (95%CI 21.3 to 26.0) of the adolescents aged 12-13 years, 21.6% (95%CI 19.7 to 23.6) aged 14-15 years, and 16.9% (95%CI 15.0 to 19.1) aged 16-17 years had high-risk HbA1c levels for diabetes. Among males and females, 23.6% (95%CI 21.8 to 25.6) and 17.5% (95%CI 15.9 to 19.2) had HbA1c values ≥5.7%, respectively. The highest prevalence was observed in Northern (23.7%; 95%CI 22.1 to 25.4) and lowest in Southern (17.6%; 95%CI 14.4 to 21.2). The prevalence of high-risk HbA1c levels was higher in Afro-descendent (27.6%; 95%CI 23.2 to 32.4) and in adolescents from public schools (21.6%; 95%CI 20.0 to 23.4) in comparison to Caucasians (16.9%; 95%CI 15.4 to 18.5) and private school students (16.7%; 95%CI 14.7 to 19.0). Also, with regard to weight status, adolescents who were obese had higher prevalence of high-risk HbA1c levels (29.7%; 95%CI 25.4 to 34.3) in comparison to adolescents who were overweight (19.7%; 95%CI 17.1 to 22.6) and eutrophics (19.3%; 95%CI 18.0 to 20.7).

Conclusion: Male sex, obesity, and lower socioeconomic status were associated with higher prevalence of high-risk HbA1c levels among Brazilian adolescents. This finding highlights the importance to focus on these priority subgroups for interventions aimed at preventing diabetes and its consequences. Socioeconomic intermediations may help to prevent diabetes in these populations.


Nothing to Disclose: GHT, RF, FVC, LAB, LGG, CK, BDS, KVB

25927 11.0000 FRI 661 A Prevalence of High HbA1c Levels Among Brazilian Adolescents Participating in the Study of Cardiovascular Risk in Adolescents (ERICA) 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Victoria Trendafilova*1, Arfana Akbar2, Irina Ciubotaru1, Hajwa Kim3, Subhash C Kukreja1 and Elena Barengolts1
1University of Illinois at Chicago, Chicago, IL, 2Jesse Brown VAMC, Chicago, IL, 3UIC Medical Center, Chicago, IL


Background: The relationship between metabolic syndrome and various risk factors has not been well-studied in African-American men (AAM).  In addition, there are limited data available on the effects of testosterone levels on cardiovascular and metabolic factors in older African-American men.

Objective: to investigate the association between metabolic syndrome (MetS) and cardiometabolic risks including serum testosterone level (sTes).

Methods: Data were from a final visit for AAM (n=178) participating in D Intervention at Veteran Administration trial (DIVA, NCT01375660). The subjects were overweight or obese AAM with prediabetes (A1C 5.7-6.4%) and Vitamin D insufficiency (25OHD=/<30ng/mL) who were treated with weekly 50,000 IU ergocalciferol for 1 year (1). MetS was defined by AHA/NHBLI criteria. Charlson index of chronic disease (Charlson) was calculated and included hypertension, cardiovascular disease (CVD), and cancer among other chronic diseases. We compared men with MetS (MetS1) and without MetS (MetS0). The linear regression models were used to test the relationship between MetS and hormonal, lifestyle and disease-related cardiometabolic risks.

Results: For the main characteristics AAM with MetS1 vs MetS0 showed (average) A1C 6.24 vs 5.97%; BMI 33.6 vs 30.6kg/m2, fat mass of 37k vs 28kg, sTes 300 vs 393 mg/dl (p<0.05 for all) and age 60 vs 58 years (p=0.08).

MetS was positively correlated with age, total body fat (TBF), android and gynoid fat, Charlson, hip circumference, perception of pain, time spent outdoors (TimeOutdoors), dietary intake of saturated fat (DietSatFat) and sTes.  In multivariable regression analysis TBF, TimeOutdoors, Charlson, DietSatFat, and sTes were selected among all risk factors that are correlated with MetS (under α=0.1) as significant (p<0.05) predictors when they are adjusting each other.

Conclusions: Modifiable lifestyle risks for MetS should be addressed in AAM with MetS. Whether testosterone treatment could be helpful in this population for preventing development of MetS and its complications including CVD and cancer remains to be determined.


Nothing to Disclose: VT, AA, IC, HK, SCK, EB

26953 12.0000 FRI 662 A Modifiable Risks Including Time Spent Outdoors and Dietary Saturated Fat As Well As Testosterone Level Are Independent Predictors of Metabolic Syndrome in African-American Men 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Hanna Savolainen-Peltonen*1, Pauliina Tuomikoski1, Pasi Korhonen2, Fabian Hoti2, Pia Vattulainen2, Mika Gissler3, Olavi Ylikorkala1 and Tomi S Mikkola1
1University of Helsinki and Helsinki University Hospital, Helsinki, Finland, 2EPID Research Oy, Espoo, Finland, 3National Institute of Health and Welfare, Helsinki, Finland


Background: In large observational studies both postmenopausal estrogen (ET) and estrogen-progestin therapy (EPT) were associated with decreased risk of cardiovascular disease. These findings were not supported by the randomized placebo-controlled trials with conjugated equine estrogens (CEE) and medroxyprogesterone acetate (MPA), initiated in women with a mean age of 63 years. A woman’s age of under 60 years at the initiation of CEE plus MPA was accompanied with vascular benefits, but an age over of 60 years with vascular risks (“window hypothesis“). Conjugated equine estrogens used in the USA may differ in vascular effects from estradiol, which is used in Europe. Moreover, various progestins, such as MPA, norethisterone acetate (NETA), dydrogesterone, or tibolone, differently affect the lipid profile, platelet function, and inflammatory markers in estrogen-using women, and therefore, progestin components of hormone therapy (HT) may be additional determinants for the cardiovascular risk. We studied the “window hypothesis” with natural estradiol alone or in combination with various progestins.

Methods: All women aged 40 years or older who had bought various systemic HT regimens or tibolone from January 1st, 1994 to the end of 2009 were extracted from the nationwide prescription register. Altogether, 498 105 women had used estradiol alone, combined with progestins (MPA, NETA, dydrogesterone, other progestins), or tibolone during 3.7 million years. The women using EPT had been most commonly exposed to NETA (48%) followed by MPA (35%), dydrogesterone (17%), and other progestins combined (17%). Women were followed from the therapy initiation to coronary heart disease (CHD) death, or to the end of year 2009. The risk of CHD death, analysed for ≤5 or >5 years of hormone use, was compared with that in the age-matched background population using standardized mortality ratio (SMR) with 95% confidence interval (CI). The data were first analysed for a 60 years of age threshold and for progestin comparisons and then, the CHD death SMR and the age at the therapy initiation were related in the entire study population.

Results: The age <60 rather than ≥60 years at the initiation of ET (SMR 0.53; CI 0.47-0.59 versus 0.76; 0.71-0.82), EPT with NETA (0.45; 0.41-0.49 versus 0.74; 0.67-0.81), or tibolone (0.35; 0.26-0.47 versus 1.01; 0.67-1.46) therapy lasting for ≤5 years was associated with significantly greater falls in the SMR. A similar tendency was also seen for other EPT groups and for longer use. In all hormone users, the CHD death SMR was smaller, the earlier the hormone therapy had started (p<0.05).

Conclusions: Our data demonstrate that the 60 years of age at the initiation of postmenopausal HT is not a threshold age, but the earlier the HT had been started, the smaller was the cardiac mortality risk. The various progestins as complements to estradiol do not modify this “timing effect”.


Nothing to Disclose: HS, PT, PK, FH, PV, MG, OY, TSM

25083 13.0000 FRI 663 A Reduced Cardiac Death Risk in Relation to the Younger Age at Initiation of Various Postmenopausal Hormone Therapies 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Adegbenga BOLANLE Ademolu*1, Abiola Olanike Ademolu2, Anthonia Ogbera3 and Augustine E Ohwovoriole4
1Lagos State University Teaching Hospital (LASUTH), Ikeja, Nigeria, 2Olabisi Onabanjo University Teaching Hospital, Ogun, Nigeria, 3Lagos State University, Nigeria, 4University of Lagos, Yaba Lagos, Nigeria


Cholesterol is an important substance in intermediary metabolism. The correlation between hyperlipidemia and hypertension, mean arterial pressure, pulse pressure is not well characterised in Africans. This paper aim to address this.This is a pilot study involving 36 hypertensives and 35 normotensives as controls. The blood pressure of each participants was measured using a mercurial sphygmomanometer at heart level  in the sitting position. The fasting plasma cholesterol for the hypertensives and control were taken using a cholesterol meter. The JNC 7TH sustained by JNC 8TH Classification of hypertension was used to define hypertension as blood pressure equal to or greater than 140/90mmHg.Hypercholesterolemia(hyperlipidemia) was  defined as fasting plasma cholesterol above 200mg/dl.The mean arterial pressure was computed by  adding diastolic blood pressure to one third of pulse pressure.The pulse pressure was computed by subtracting diastolic from systolic blood pressure.All measurements were taken during a free  medical screening programme in Surulere Lagos,Nigeria,West Africa.The age range among  hypertensives was 30 to 81years while it was 18 to 88years among normotensive controls. Hyperlipidemia  was present in 33.3% of the hypertensives while it was present in only 17.1% of the control.We can therefore infer that in this African cohort sample studied though small in  size, hyperlipidemia is twice as common in hypertensives in Africa compared to the general African population. Genderwise,hyperlipidemia was commoner among  Hypertensives that are  female. Further analysis showed that of the 18 subjects (25.35%) in the study with hyperlipidemia ,66.6% had hypertension while 33.3% were normotensive. Routine screening of hypertensives for hyperlipidemia is supported by this study as 1 out of 3 hypertensives in Africa have hyperlipidemia. This paper findings also support the routine screening for hypertension among subjects with hyperlipidemia as 2 out of 3 with hyperlipidemia  had hypertension. There is no correlation between hyperlipidemia and mean arterial pressure in hypertensive Africans according to this study.Non hypertensive Africans with hyperlipidemia tend to have lower mean arterial pressure than the other non hypertensive general population without hyperlipidemia.Hyperlipidemia occurs commonly at  low pulse pressure in hypertensives and at  wide pulse pressure in non hypertensive Africans.There is correlation between hyperlipidemia and hypertension and pulse pressure but none between hyperlipidemia and mean arterial blood pressure in African hypertensives according to this pilot  study.

Nothing to Disclose: ABA ,AA,OA,AO                                                                                                                                                                                                                                                                                                                                                                                                                      


Nothing to Disclose: ABA, AOA, AO, AEO

24204 14.0000 FRI 664 A Correlation Between Hyperlipidemia and Hypertension,Mean Arterial Pressure,Pulse Pressure Among Africans 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

James Carey, Julia Wenger, Jennifer Huynh, Ravi Thadhani and Rhonda Bentley-Lewis*
Massachusetts General Hospital, Boston, MA



Gestational diabetes mellitus (GDM) is associated with increased maternal cardiovascular disease (CVD) risk manifest as hypertension subsequent to pregnancy (1, 2). The impacts of dysglycemia and race/ethnicity on hypertension risk have been examined (3); however, the study of other putative risk factors has been limited.


We hypothesize that GDM and gestational dysglycemia are sufficiently predictive of hypertension subsequent to pregnancy and that a risk score based on degree of dysglycemia can be used to identify at-risk patients.


We examined a cohort of 23,223 women who presented for prenatal care to the Massachusetts General Hospital Obstetrical Department between September 1998 and January 2007. We selected women, ages 18 to 40 years, free of CVD or pre–GDM, with complete glycemic and demographic data who delivered a singleton, live birth. Each woman’s initial pregnancy was selected in order to not violate the independence assumption. The population was stratified by incident hypertension subsequent to pregnancy. Logistic regression models (with and without stepwise selection) were used to identify significant maternal risk factors based on their predictive performance. Finally, the risk prediction score was developed from β-coefficients in multivariable Weibull models.


The study sample comprised of 11,161 women; 547 (4.9%) women developed hypertension during 4.2 years (median) follow-up. Significant predictors of hypertension included age, body mass index, systolic and diastolic blood pressures, parity, dysglycemia, and race/ethnicity. These factors were used to develop a risk score for subsequent hypertension risk. The final risk score achieved a c-statistic of 0.76, indicating moderate discriminative ability.


We developed a risk score that may be used to identify patients with an elevated risk of hypertension subsequent to a pregnancy complicated by dysglycemia. Further external validation is warranted to determine clinical effectiveness.


Nothing to Disclose: JC, JW, JH, RT, RB

25013 15.0000 FRI 665 A Development of a Risk Score for Predicting Hypertension in Women with a History of Gestational Diabetes Mellitus 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Ramon Bossardi Ramos*1, Vitor Costa Fabris2, Maria Augusta Maturana3 and Poli Mara Spritzer4
1Hospital de Clínicas de Porto Alegre (HCPA), Porto Alegre, RS, Brazil, 2Universidade Federal Rio Grande do Sul, 3Gynecological Endocrinology Unit, Division of Endocrinology, Hospital de Clínicas de Porto Alegre, 4Hospital de Clínicas de Porto Alegre and Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil


Postmenopausal women experience an increase in cardiovascular disease (CVD), which could be associated with both aging and the changing hormonal status. Evidence suggests that the development of CVD results from environmental and genetic factors. Epigenetics, including DNA methylation seems to present major relevance to all disease processes, including CVD and related conditions. Therefore, the aim of the present study was to determine whether global DNA methylation was associated with cardiovascular risk in postmenopausal women. In this cross sectional study carried out in a university hospital, 90 postmenopausal women (55.5±4.9 years, 5.8 (3.0–10.0) years since menopause) were enrolled. The inclusion criteria were menopause (defined as follicle-stimulating hormone levels above 35 IU/L and last menstrual period at least 1 year before the beginning of the study), age between 45 and 65 years, and no use of hormone therapy for at least 3 months before the enrollment. Exclusion criteria were prior diagnosis of CVD, current smoking or a diagnosis of diabetes. DNA was extracted from peripheral leukocytes and global DNA methylation levels were obtained with an ELISA kit (MDQ1, Imprint® Methylated DNA Quantification Kit, Sigma Aldrich). Cardiovascular risk was estimated by using the Framingham General Cardiovascular Risk Score (10-year risk) (FRS). All participants were evaluated for clinical and laboratorial variables. Patients were stratified into 2 groups according to FRS, low cardiovascular risk (FRS: <10%, n=69) and medium/high cardiovascular risk (FRS ≥10%, n=21). Age, time since menopause, blood pressure, total cholesterol and LDL-c levels were higher in FRS ≥10% group in comparison with the FRS <10% group. BMI, triglycerides, HDL-c, HOMA-IR, glucose and C-reactive protein levels were similar between the two groups. Global DNA methylation (% 5mC) in the overall sample was 26.5% (23.6 – 36.9). The FRS ≥10% group presented lower global methylation levels compared with FRS <10% group: 23.9% (20.6 – 29.1) vs. 28.8% (24.3 – 39.6), p= 0,02. This analysis remained significant even after adjusting for time since menopause (p= 0.02).  In conclusion, our results indicate global DNA hypomethylation is associated with higher cardiovascular risk in postmenopausal women and support further endeavors to assess epigenetic mechanisms for CVD in this population in future large-scale epidemiological studies.


Nothing to Disclose: RBR, VCF, MAM, PMS

26758 16.0000 FRI 667 A Association Between Global Leukocyte DNA Methylation and Cardiovascular Risk in Postmenopausal Women 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Korapat A Mayurasakorn, Tsuyoshi Homma, Mika Homma, Isis Akemi Katayama*, Tham Yao, Jose R Romero, Gail K. Adler, Gordon H Williams and Luminita H Pojoga
Brigham and Women's Hospital, Harvard Medical School, Boston, MA


Background and Purpose—Caveolin-1 (CAV-1), a plasma membrane protein has been shown to be involved in modulating glucose homeostasis and the actions of the renin-angiotensin-aldosterone system (RAAS). Caloric restriction (CR) is widely accepted as an effective therapeutic approach to improve insulin sensitivity and reduce the severity of diabetes. Recent data indicated that polymorphisms of the CAV-1 gene are strongly associated with insulin resistance, hypertension and metabolic abnormalities in non-obese phenotypes. Therefore, we determined whether the CR improves the abnormal metabolic phenotypes and several cardiovascular (CV) risk factors in the lean CAV-1 KO mice.

Materials/Methods- Twelve- to fourteen-week-old cav-1 knockout (KO) and genetically matched wild-type (WT) male mice were subjected to 2 dietary regimens: ad libitum (ad lib) or paired-fed 40% CR for 4 weeks. Three weeks following the onset of dietary restriction, all groups were assessed for insulin sensitivity. At the end of the study, all groups were assessed for fasting glucose, insulin, HOMA-IR, lipoproteins, corticosterone levels and blood pressure. Aldosterone secretion was determined from acutely isolated Zona Glomerulosa cells. Renal mRNA expression of serum-and glucocorticoid-inducible kinase 1 (SGK1) and the epithelial sodium channel (ENaC) were measured by quantitative real time RT-PCR.

Results-We confirmed that the CAV-1 KO mice display a phenotype consistent with the cardiometabolic syndrome, as shown by higher systolic blood pressure (SBP), plasma glucose, HOMA-IR and aldosterone levels despite lower body weight. CAV-1 KO mice maintained their body weight on the ad lib diet, but had substantially greater weight loss with CR, as compared to WT mice. CR-mediated changes in weight were associated with dramatic improvements in glucose and insulin tolerance in both genotypes. These responses, however, were more robust and were related to reduced plasma insulin, and HOMA-IR in CAV-1 KO vs. WT mice. Surprisingly, in the CAV-1 KO vs. WT mice, CR was associated with increased aldosterone levels which correlated with an increase in SGK1 and ENaC mRNA expression, which may explain the net increase in sodium retention and the BP response to CR in the CAV-1 KO. This suggests that in these mice CR induced an increase in CV risk.

Conclusions—CR improved the metabolic phenotype in CAV-1 KO mice by increasing insulin sensitivity; nevertheless, this intervention also increased CV risk. The underlying mechanism may be related to inappropriate aldosterone regulation potentially leading to exaggerated sodium retention in the kidney and ultimately to blood pressure increase.


Nothing to Disclose: KAM, TH, MH, IAK, TY, JRR, GKA, GHW, LHP

26666 17.0000 FRI 668 A Caloric Restriction: A Paradox of Improved Glucose Homeostasis Vs. Hyperaldosteronism, Leads to Increased Cardiovascular Risk in Caveolin-1-Deficient Mice 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Hyeong Kyu Park*1 and Mi Kyung Kwak2
1Soonchunhyang University Hospital, Seoul, Korea, Republic of (South), 2Soonchunhyang Univ. Hospital, Seoul, Korea, Republic of (South)


Angiopoietin-like protein (ANGPTL) 6, a novel hepatokine, is known to modulate angiogenesis and metabolism. ANGPTL6 has been shown to be higher in metabolic syndrome than in healthy individuals. Recent clinical data suggest a potential role of ANGPTL6 in endothelial dysfunction. However, the functional role of ANGPTL6 in type 2 diabetes mellitus (T2DM) has not been determined. Therefore, we analyzed plasma ANGPTL6 levels and other biochemical markers in patients with T2DM.

A total of 108 Korean patients with T2DM were enrolled. Subjects with known cardiovascular diseases, chronic kidney disease, or active infection were excluded. Plasma ANGPTL6 was quantified. We assessed vascular health status by measuring carotid intima-media thickness (IMT).

Plasma ANGPTL6 correlated positively with C-reactive protein (CRP; r = 0.31, P < 0.01) and resistin (r = 0.28, P < 0.05), and negatively with high-density lipoprotein (HDL) cholesterol (r = - 0.24, P < 0.05). However, ANGPTL6 did not correlate with carotid IMT, adiponectin, or leptin. A multiple regression analysis showed that CRP and HDL cholesterol remained independently associated with ANGPTL6 after adjustment for age, gender, body mass index, and resistin.

Circulating ANGPTL6 concentrations are positively associated with an inflammatory marker, and negatively with HDL cholesterol in subjects with T2DM. Further studies will be needed to explore the physiologic functions of ANGPTL6 in T2DM.


Nothing to Disclose: HKP, MKK

27380 18.0000 FRI 670 A Angiopoietin-like Protein 6 Is Associated Wth an Inflammatory Marker and Low HDL Cholesterol in Type 2 Diabetes 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Sirimon Reutrakul1, Nantaporn Siwasaranond*1, Hataikarn Nimitphong2, Sunee Saetung3, Naricha Chirakalwasan4, La-or Chailurkit5, Kriangsuk Srijaruskul6, Boonsong Ongphiphadhanakul7 and Eve Van Cauter8
1Faculty of Medicine Ramahibodi Hospital, Bangkok, Thailand, 2Department of Medicine, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok, Thailand, 3Department of Medicine, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand, 4Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand, 5Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand, 6Faculty of Medicine Ramathibodi Hospital, Bangkok, Thailand, 7Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok, Thailand, 8University of Chicago, Chicago, IL


Context Melatonin, a hormone released during the biological night under control by the circadian clock, plays a role in modulating the sleep-wake cycle. Reduced nocturnal melatonin release has adverse effects on glucose metabolism and promotes the risk of type 2 diabetes (T2DM). Obstructive sleep apnea (OSA) is a common comorbidity of type 2 diabetes (T2DM) that has been associated with poorer glycemic control. Whether nocturnal urinary excretion of 6-sulfatoxymelatonin in type 2 diabetes is affected by the presence and severity of OSA is unknown.

Objective To explore the relationship between nocturnal urinary excretion of 6-sulfatoxymelatonin and OSA severity in type 2 diabetes

Setting and Design A cross-sectional study conducted at the endocrinology clinic, Ramathibodi Hospital, Mahidol University, Bangkok, from March to December 2014

Materials and methods Fifty-six patients with type 2 diabetes who were not taking beta-blockers, had not been previously diagnosed with OSA and were not shift workers were recruited.  Participants underwent 7 nights of ambulatory monitoring of habitual sleep duration and fragmentation by actigraphy. On one of these nights, the presence of OSA was assessed using a validated home monitoring device (WatchPAT-200). Urinary 6-sulfatoxymelatonin was measured on an overnight urine sample. The ratio of 6-sulfatoxymelatonin to creatinine was used as an indicator of nocturnal melatonin release.

Results: OSA was diagnosed in 43 participants (76.8%). The presence of OSA was associated with a lower ratio of 6-sulfatoxymelatonin to creatinine [9.7 (5.2-17.2) ng/mg vs 17.8 (9.7-28.1) ng/mg, p=0.05). Longer diabetes duration, retinopathy, insulin use and higher HbA1c correlated with lower 6-sulfatoxymelatonin excretion. Greater OSA severity [as assessed by apnea hypopnea index (AHI), oxygen desaturation index (ODI) and percentage of sleep time spent under oxygen saturation of 90% (T90)] was significantly correlated with lower urinary excretion of 6-sulfatoxymelatonin, while sleep duration and fragmentation were not. After adjusting for diabetes duration, retinopathy, insulin use and HbA1c, ODI (B= -0.331, p=0.015) and T90 (B= -0.224, p=0.001) independently predicted lower nocturnal urinary excretion of 6-sulfatoxymelatonin.

Conclusion OSA severity was independently associated with lower nocturnal urinary excretion of 6-sulfatoxymelatonin in T2DM patients, suggesting a negative interaction between overnight melatonin release and OSA. Whether melatonin supplementation will be beneficial in patients with T2DM and OSA is an open question worthy of future research.


Disclosure: SR: Speaker, Sanofi, Speaker, Medtronic Minimed, Clinical Researcher, Merck & Co.. EV: Clinical Researcher, Shire/Viropharma, Clinical Researcher, Philips/Respironics, Clinical Researcher, Astra Zeneca, Clinical Researcher, Amylin Pharmaceuticals, Consultant, Shire/Viropharma, Consultant, Vanda Pharmaceuticals, Consultant, Philips/Respironics. Nothing to Disclose: NS, HN, SS, NC, LOC, KS, BO

24123 19.0000 FRI 671 A The Relationship Between Nocturnal Urinary Excretion of 6-Sulfatoxymelatonin and Obstructive Sleep Apnea Severity in Type 2 Diabetes 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Iris Gutmark-Little*1, Dhananjay Subramaniam2, Goutham Mylavarapu3, William Stoddard2, Ephraim Jeff Gutmark2, Christian Trolle4, Steffen Ringgaard5, Philippe Backeljauw1, Kristian Havmand Mortensen6 and Claus H. Gravholt4
1Cincinnati Children's Hospital Medical Center, Cincinnati, OH, 2University of Cincinnati, 3Cincinnati Children's Hospital Medical Center, 4Aarhus University Hospital, Aarhus, Denmark, 5Aarhus University, 6Cambridge University Hospitals, Cambridge, United Kingdom


Background: Change in aortic dimension in Turner syndrome (TS) is poorly described, despite dilatation being a key risk factor for dissection. We have developed a computed methodology that enables continuous aortic measures for improved monitoring of progressive disease.

Objective and hypotheses: Continuous aortic measures will improve reproducibility, accuracy, and robustness of dimension quantification. Our objective was to use patient-specific geometries to validate this tool and show advantages over an existing method.

Methods: ECG-triggered, static, diastolic longitudinal cardiac magnetic resonance (CMR) scans for 15 TS subjects (age = 49 ± 6.4 years; 2-4 scans/subject/5-10 years; 44 total scans).

3D aortic geometries were reconstructed, defining centerlines for each. Cross-sectional area and maximum (max) diameter were computed continuously, perpendicular to this line (aortic root to diaphragm). Available manual measurement max diameter data at 9 defined locations was compared and correlations calculated. To assess variability between methods, % aortic growth/time and standard deviation (SD) were calculated per segment (ascending - AA, transverse - TA, descending - DA). Aortic growth was defined as change beyond inherent continuous method variability.

Results: Manual and continuous measures were highly correlated (r-value 0.77, p <0.01).

Less variability was shown using the continuous method (SD of % change between scans: AA/Computed 1.95, AA/Manual 8.21, p <0.01; TA/C 2.5, TA/M 5.7, p <0.01; DA/C 1.8, DA/M 8, p <0.01).

12/15 had no change in continuous dimensions over time, despite 7/12 showing change using manual measurements. 3/15 subjects showed aortic growth; 2 of which showed agreement between methods.

Conclusions: Continuous automated measurement ensures rapid, accurate, and reproducible detection of changes over the entire aortic span.

The advantages of this method are:

  1. Reproducibility and less user bias. Eliminates the need for measurement at identical anatomic location and cross-sectional tilt angle.
  2. Complete anatomic information throughout aortic span, optimizing measurements near steep gradients (i.e. coarctation or near branch points, where manual overestimation occurs).
  3. Neighbouring points establish continuity and validation.
  4. Robustness of data, assessing the entire aortic geometry (i.e. max diameter and circumferential location, area, aspect ratio, curvature, tortuosity). True max diameter is identified given the full circumferential cross-sectional information.
  5. CMR artifacts identified due to curve continuity.

Most subjects had stable aortic dimensions over time using continuous measures after several had been noted to have growth by manual measurements, decreasing unnecessary intervention. This tool allows endocrinologists, as primary TS providers, improved identification of those with progressive disease.


Disclosure: PB: Advisory Group Member, Novo Nordisk, Advisory Group Member, Sandoz, Advisory Group Member, EMD Serono, Advisory Group Member, Ipsen. Nothing to Disclose: IG, DS, GM, WS, EJG, CT, SR, KHM, CHG

24398 20.0000 FRI 672 A Monitoring for Aortic Dilatation Using Continuous Aortic Dimensions in Turner Syndrome 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Jan Wen*1, Christian Trolle2, Steffen Ringgaard3, Niels Holmark Andersen2, Iris Gutmark-Little4, Ephraim Jeff Gutmark5, Kristian Havmand Mortensen6 and Claus H. Gravholt2
1Aarhus University Hospital, Aarhus C, Denmark, 2Aarhus University Hospital, Aarhus, Denmark, 3Aarhus University, 4Cincinnati Children's Hospital Medical Center, Cincinnati, OH, 5University of Cincinnati, 6Cambridge University Hospitals, Cambridge, United Kingdom



A high prevalence of cardio-vascular malformations and non-structural cardio-vascular diseases characterizes individuals with Turner Syndrome (TS). The incidence of aortic dissection is 100 fold increased, and cardio-vascular disease explains 50% of the excess mortality. Reduced aortic wall distensibility is associated with increased risk of cardio-vascular diseases in the general population and may bear impact on the risk of aortic dissection. A prior study (van den Berg et al 2006) has shown a significantly lower aortic distensibility of the ascending aorta at the level of the pulmonary bifurcation and of the descending aorta at the level of the diaphragm, compared to age-matched and BSA-matched female controls. We posit that aortic distensibility is reduced in TS patients compared to controls and associated with an increase in Pulse Wave Velocity (PWV) and the risk of aortic dilatation. We, for the first time, relate these measures to the changes in aortic diameter during a 4.8 year follow-up.

Material and methods

Participants were recruited from an ongoing prospective study, and we performed a cross-sectional study assessing aortic distensibility in relation to progression in aortic diameter. The study, at baseline, included 102 TS individuals and 67 age-matched 46,XX controls. 90 TS individuals and 67 controls were asked to participate again. As of today, 33 with TS (47.6 years, range [27-62]) and 25 controls (51.3 years, range [26-67]) have participated. Aortic distensibility was assessed with MRI (Philips Achieva-dStream 1.5 Tesla) at three different positions: 1) The ascending aorta at the pulmonary artery bifurcation, 2) The aortic arch, and 3) The descending aorta at the level of the pulmonary artery bifurcation. Images were analyzed using Siswin®. Blood pressure (Aneroid Sphygmomanometer, ERKA, Germany), Pulse Wave Velocity (SphygmoCor, AtCor Medical, Australia) were measured. Physical examination, 24-hour ambulatory blood pressure, and medical history were done as well.


Our principal finding is an increase in aortic distensibility of borderline significance (P = 0.0505) in TS compared to controls. We find comparable aortic distensibility within TS subgroups: Tricuspid aortic valve vs. Bicuspid aortic valve (P = 0.1); 45,X vs. mosaics (P = 0.7); Elongated Thoracic Aorta (ETA) vs. non-ETA (P = 0.2); dilatation vs. no dilatation (P = 0.6). In addition, we find no difference in PWV between TS and controls (P = 0.2).


Contrary to previous studies, our preliminary results suggest a trend towards an increase in aortic distensibility at the level of the aortic arch. We speculate that this discrepancy is due to an older study population.


Nothing to Disclose: JW, CT, SR, NHA, IG, EJG, KHM, CHG

26493 21.0000 FRI 673 A Aortic Distensibility in Turner Syndrome Compared to Sex-Matched Controls with the Use of MRI 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Fiona Dore1, Aytan Mammadova2, Richard Amdur3 and Sabyasachi Sen*4
1George Washington University Medical Faculty Associates, 2George Washington University, 3Georgetown University School of Medicine, 4The George Washington University, DC


Endothelial dysfunction leading to vascular complication is a major concern in a type 2 diabetes (DM) and obese population. Arterial stiffness is an established predictor of endothelial health and cardio-vascular disease (CVD) risk. Arterial stiffness is commonly measured by two tonometry based tests: pulse wave velocity (PWV) and pulse wave analysis (PWA), which provides the values of Augmentation index-75 (AI-75) and Augmentation Pressure (AP). Serum biochemistry is also used to monitor disease progression. In this cross-sectional study, we wanted to determine which biochemical or biophysical measure is the best overall predictor of arterial stiffness. The measures investigated were: C-Reactive Protein (CRP), IL-6, TNF-alpha, Leptin, Adiponectin, Insulin, Glucose, LDL, Non-HDL Cholesterol, BMI, Total Fat Mass, Fat Free Mass, % fat, % body water, waist circumference, hip circumference, and waist/hip ratio with PWV, AI-75, and AP.

Data from 16 subjects with Diabetes Mellitus ≤ 8 years, age 40-70 years, with HbA1c 6.0 – 9.0, and a BMI of 25 – 39.9 were analyzed.

Looking at the Pearson correlation between individual biochemical and biophysical measures, and the individual measures of Arterial Stiffness (PWV, AI-75, AP), the results show that CRP is very strongly correlated to PWV (p=0.6), and moderately correlated to AP and AI-75 (p=0.44, 0.30). Leptin is found to be relatively equally well correlated to PWV, AI-75, and AP (p=0.30, 0.34, 0.30). However, when looking at a composite AS score (from standardized versions of PWV, AI-75, and AP) versus the biophysical and biochemical measures, the only measures of significance were CRP, Fat Mass, and Hip measurements. When these values were placed in a multivariate model, the only independent predictors of AS were Fat Mass (p=.018), and Hip measurements (p=0.016). When taking the numerous measures of AS into account, it is found that Fat Mass and Hip measurements were the single best predictors of arterial stiffness, even better than any of the serum biochemistry parameters. In conclusion, if we want to determine a single factor which has a the most bearing on a type 2 diabetes subjects arterial stiffness, then degree of fat mass seems to be the best predictor even better than any of the biochemical parameters. Therefore, in our study, fat mass appears to be an important indicator of CVD risk.


Nothing to Disclose: FD, AM, RA, SS

24994 22.0000 FRI 674 A Fat Mass Is the Best Single Predictor of Arterial Stiffness in a Type 2 Diabetes Population 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Jessica A. Lundgren*, Paul V. Targonski, Charles H. Brooks, Jamie L. Kennedy and Jennifer L. Kirby
University of Virginia, Charlottesville, VA


Background: Systolic heart failure (SHF) and type 2 diabetes mellitus commonly co-exist, with each disease process exacerbating the detrimental effects of the other. SHF may worsen diabetes control due to decreased patient exercise tolerance, decreased cardiac output with consequent reduction in skeletal muscle glucose uptake, and increased insulin resistance related to elevated cortisol and catecholamines. Increasingly, left ventricular assist devices (LVADs) are being implanted to provide circulatory support in advanced SHF. Several small studies suggest that diabetes control improves after LVAD placement.

Objective: Our primary objective was to study whether diabetes control improved after LVAD placement in the immediate post-operative setting and in longer-term outpatient follow up. Our secondary objective was to investigate factors that may relate to change in diabetes control.

Methods: Single-center, retrospective study of 47 adult patients with established diagnoses of SHF and diabetes mellitus who underwent LVAD placement from 1/1/2009 to 10/20/2013 at the University of Virginia and survived to discharge. Primary endpoints were change in daily insulin dose from admission to discharge, and change in hemoglobin A1c (HbA1c) from pre-LVAD to follow up. When multiple post-LVAD HbA1c values were available, we used those collected closest to 180 days post-LVAD. Secondary endpoints were change in creatinine from admission to 1 month post-LVAD, and change in BMI from pre-LVAD to 1 month post-LVAD.

Results: In patients receiving insulin injections (n=20), daily insulin dose decreased 45% from admission to discharge (95% CI, -65% to -26%; p<0.0001). Average absolute change in HbA1c was -1.81% (n=41, 95% CI, -2.38 to -1.26%; p<0.0001), at an average of 233 days post-LVAD. By linear regression, HbA1c decreased significantly as BMI decreased (n=47, F=6.74, p=0.013). Creatinine decreased by an average of -0.59 mg/dL at 1 month post-LVAD (n=47, 95% CI, -0.79 to -0.40 mg/dL; p<0.001); this was not significantly related to HbA1c improvement, however, there was a trend towards improvement.

Conclusions: We observed significant decreases in insulin requirements by the time of post-LVAD discharge and significant decreases in HbA1c over longer-term follow up. Our findings demonstrate durable improvement in glycemic control after LVAD implantation. There are several potential explanations for improved glycemic control, including reduced systemic inflammation, improved renal function and decreased BMI. Further studies are needed to investigate the physiologic mechanisms underlying the improvement in diabetes control.


Nothing to Disclose: JAL, PVT, CHB, JLK, JLK

25307 23.0000 FRI 675 A After Left Ventricular Assist Device Placement in Advanced Heart Failure, Diabetes Control Improved Immediately and in Outpatient Follow up 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Katie L O'Sullivan*1, Magdalena Dumin2, Leila Kheirandish-Gozal1, Rakesh Bhattacharjee3, David Gozal1 and Dorit Koren1
1University of Chicago, Chicago, IL, 2University of Chicago Medical Center, Chicago, IL, 3Rady's Children's Hospital, San Diego


Background: Acute sleep deprivation is associated with obesity and type 2 diabetes (T2DM) in adults. Chronic insufficient sleep associates with obesity in children, but other adverse cardiometabolic sequelae of acute sleep deprivation such as T2DM risk are less well-described in children.

Objective: To examine the metabolic impact of acute sleep restriction in obese adolescents.

Design/Methods: Seven obese, pubertal adolescents (15.4±2.1 yrs, BMI 36.4±5.9 kg/m2; 86% female, 100% African-American) enrolled in a pilot study. Habitual sleep duration was recorded by wrist actigraphy over 2 weeks. Participants restricted sleep duration by 1 hour/night for 1 week. Subjects underwent anthropometric measurements and a 180-minute oral glucose tolerance test (OGTT) with ghrelin and leptin levels at baseline and after sleep restriction.

Results: Participants spent 1.5 hours less in bed (8.9±0.8 versus 7.4±1.0 hours; p=0.002) and slept 1 hour less (6.9±0.8 versus 5.9±0.6 hours; p=0.008) during the week of sleep restriction compared to baseline. Participants gained an average of 0.83 kg (from 99.6±13.8 to 100.4±14.1 kg; p=0.063) over the week of sleep restriction. 30-minute post-challenge glucose levels were significantly higher and 15-minute post-challenge glucose levels trended toward being higher after sleep restriction; 15-minute post-challenge insulin levels trended higher in the sleep-restricted state. Mean leptin levels were higher following sleep restriction (79.4±46.8 versus 68.8±41.3 ng/mL, p=0.04). 120-minute ghrelin levels trended higher in the sleep-restricted state (54.0±18.1 versus 37.4±5.2 pg/mL, p=0.094).

Conclusions: In our pilot group, participants gained weight and had mildly higher post-challenge glucose and insulin levels following short-term, mild sleep restriction; leptin levels were higher, contrary to what is seen in adults but consistent with prior studies in children. We speculate that leptin levels may indicate greater leptin resistance and higher ghrelin may mediate greater post-prandial hunger, potentially mediating weight gain. Optimizing sleep duration may reduce weight gain and potentially glucose levels in obese adolescents. Our preliminary findings warrant expansion of our pilot study to a larger cohort.


Nothing to Disclose: KLO, MD, LK, RB, DG, DK

25812 24.0000 FRI 676 A The Effects of Acute Sleep Restriction on Weight and Cardiometabolic Risk Indices in Obese Adolescents 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Irina Dzherieva*1, Natalya Volkova2, Ilya Davidenko3, Maria Porksheyan4, Igor Reshetnikov5, Aida Gulmagomedova6 and Maria Komurdzhyantz7
1Rostov State Medical University, Rostov on Don, Russia, 2The Rostov State Medical University, Rostov-on-Don, Russia, 3Rostov State Medical University, 4Rostov State Medical University, Rostov-on-don, 5Rostov State Medial University, Rostov on Don, Russia, 6Rostov State Medical University, Rostov on Don, Russia, 7Hospital #7 Rostov on Don Russia, Rostov on Don, Russia


Shift work is a risk factor of dislipidemia and diabetes melitus. There is a hypotesis that melatonin (M) might be a link between shift work and these metabolic desoders. To study the influence of shift work on the secresion of M and the development of dislipidemia and diabetes melitus  we observed two groups: group «A» including  26 men, avarage age 44±4.3 (95%CI:41.6-47.0)years,   with shift work more 6 years and night work once a week and  group «B» including 25  men avarage age 45.1±2.3 (95%CI:41.1-46.1)years who had day-time work only. High-density lipoproteins (HDL), fasting triglycerides (TG), fasting glucose were determined. The secretion of M was determined according to excretion 6-sulfatoxymelatonin (MT6S) in urine.   The total MT6S in both groups was equal, p=0.07759. The  night - time MT6S in group «A» (23.6 ±16.3 95% CI: 16.9-31.1) ng/ml was less than in group «C», p=0.028. The day –time MT6S in group «A»(10.2±6.9 (95%СI: 7.3-13.1) ng/ml was higher than in group «C», p<0.001. The peak of MT6S was less in group «A», p=0.014 although the peak was observed in both groups. In group «A» HDL were 0.96±0.23 (95% CI:1.04-1.29) mmol/L vs 1.30±0.08 (95% ДИ 1.27-1.35) mmol/L, р=0.030 in group«C». In group «A» TG  were 2.4±1.3 (95% CI:0.4-3.9) mmol/L higher than in group «C» р=0.046.  The correlation was determined between the peak of MT6S and fasting TG (r= - 0.34), HDL (r=0.26), р<0.05 and fasting glucosae(r=-0.30, p<0.05) .  The  night - time MT6S and the day – time MT6S correlated with HDL (r=-0.24 и r=-0.40 respectively, р<0.05). The falling of the peack of MT6S leaded to increasing of  hypertriglyceridemia (OR=1.4, 95% CI: 0.7–2.1, p<0.05), declining of HDL (OR=1.7, 95% CI: 0.9–2.6, p<0.05), hyperglicemia  (OR=8.0, 95% CI: 6.1–10.2, p<0.05). The decreasing night - time MT6S leaded to raising of hypertriglyceridemia (OR=1.4 95% CI: 0.5–3.7, p<0.05), declining of HDL (OR= 1.4.95% CI: 0.5–3.7, p<0.05), hyperglicemia (OR=7.7, 95% CI: 5.1–10.8, p<0.05). The increasing day –time MT6S leaded to reducing of HDL (OR=1.6, 95% СI: 0.5–3.7, p<0.05). Thus shift work leaded to desoders of M secrecion andthis prosses in its turn promoted  dislipedemia and diabetes melitus.


Nothing to Disclose: ID, NV, ID, MP, IR, AG, MK

26896 25.0000 FRI 677 A Disoder of Melatonin Secretion and Shift Work 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Omar Bayomy*1, Ajay D Rao2, Rajesh K Garg2, Anand Vaidya3, Beata Reiber2, Stephanie Nijmeijer2, Michael Jerosch-Herold2, Raymond Y Kwong2 and Gail K. Adler3
1Harvard Medical School, Boston, MA, 2Brigham and Women's Hospital/Harvard Medical School, Boston, MA, 3Brigham and Women's Hospital, Harvard Medical School, Boston, MA



Plasminogen activator inhibitor-1 (PAI-1), a pro-thrombotic and pro-inflammatory protein, is implicated in the pathophysiology of coronary artery disease (CAD).  PAI-1 levels are increased in type 2 diabetes mellitus (T2DM).  Epicardial fat is a CAD and cardiovascular disease risk factor and correlates with PAI-1 in non-diabetic obese individuals.  The relationship between epicardial fat and PAI-1 in individuals with T2DM is uncertain.  We tested the hypothesis that epicardial fat predicts PAI-1 levels independent of body mass index (BMI) in patients with well-controlled T2DM.


We studied thirty-seven study participants (25 male, 12 female, ages 35-70 years) with T2DM, without clinical evidence of CAD or other active medical problems except for hypertension.  Study participants underwent a 3 month run-in period during which medications were adjusted to achieve good cardiometabolic control.  Study participants then underwent cardiovascular magnetic resonance imaging.  Axial T1 turbo spin echo images were analyzed to quantify epicardial fat volume by software tracing using QMassMR7.6 software.  Partners HealthCare Institutional Review Board approved the protocol, and all participants provided written informed consent.


At the end of the 3 month run-in period, hemoglobin A1c (HbA1c) was 7.07 ± 0.87% [mean ± standard deviation]; BMI 32.66 ± 3.79 kg/m2; average systolic and diastolic blood pressures 127.9 ± 14.48 and 76.26 ± 8.95 mmHg, respectively; cholesterol 147.51 ± 33.42, triglycerides 119.43 ± 61.73, HDL 40.05 ± 10.87, cVLDL 23.86 ± 12.38, and cLDL 83.59 ± 28.37 [all mg/dL]; PAI-1 4.56 ± 2.4 ng/mL. 

Both epicardial fat and BMI correlated with PAI-1 (R2 = 0.17, p = 0.01; R2 = 0.28, p < 0.001, respectively) in univariate analyses.  In multivariable regression analysis (model included epicardial fat and BMI), both epicardial fat and BMI were independent predictors of PAI-1 (epicardial fat, T = 2.11, P = 0.04; BMI, T = 3.21, P = 0.003).  There was no significant correlation between epicardial fat and BMI (R2 = 0.06, p = 0.14) and PAI-1 was not associated with age, gender or HbA1c. 


Epicardial fat is significantly related to PAI-1 in patients with T2DM independent of BMI.  This result suggests that in diabetes, as shown previously in non-diabetic populations, epicardial fat contributes to pro-inflammatory pathology.


Nothing to Disclose: OB, ADR, RKG, AV, BR, SN, MJ, RYK, GKA

25992 26.0000 FRI 678 A Epicardial Fat and Plasminogen Activator Inhibitor-1 Levels in Patients with Type 2 Diabetes Mellitus 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Ibiye Owei*1, Nkiru Umekwe1, Hanan Mohamed1, Sotonte Ebenibo1, Jim Wan2 and Samuel Dagogo-Jack1
1University of Tennessee, Memphis, TN, 2The University of Tennessee Health Science Center, Memphis, TN


Background:Endothelial function (EF) predicts cardiovascular risk but its interactions with race/ethnicity and glycemic status remain to be clarified.

Subjects and Methods:We assessed EF in relation to glycemia and cardiometabolic profile in African American (AA) and European American (EA) offspring of parents with type 2 diabetes (T2DM), who are participants in the prospective Pathobiology and Reversibility of Prediabetes in a Biracial Cohort (PROP-ABC) study. Assessments at enrollment included a 75-g OGTT, blood pressure and anthropometric measurements, lipid profile and blood biochemistries. Other assessments during subsequent visits included insulin sensitivity and secretion, body composition (dexa) and resting energy expenditure (REE). Endothelial function was assessed using EndoPAT 2000 (Itamar medical) and expressed as Reactive Hyperemia Index (RHI).

Results: We studied 190 subjects (100 AA, 90 EA), mean (± SD) 53.1 ± 9.1yrs and BMI 30.6 ± 6.8 kg/m2. Based on OGTT data, 96 subjects (52 AA, 44 EA) had prediabetes and 94 subjects were normoglycemic (46 C and 48 AA). The RHI tended to be lower in AA than EA offspring (2.17 ± 0.55 vs 2.36 ± 0.72), but was similar in men and women (2.19 ± 0.64 vs 2.29 ± 0.65, P>0.05). Compared to normoglycemic controls, prediabetic subjects had lower RHI (2.14 ± 0.62 vs. 2.38 ± 0.65, P=0.013). Using linear regression, RHI was significantly correlated with age (R= 0.21, P=0.005) and weight (R= -0.16, P=0.03) in all subjects, with total cholesterol (P=0.02) in men but not women, and with 2hr plasma glucose (P=0.04) in AA but not EA subjects.  No significant correlations were seen between RHI and blood pressure, ankle brachial index, and REE.

Conclusion: In our biracial cohort, endothelial dysfunction was a sensitive indicator of recent transition from normoglycemia to prediabetes status. This finding reinforces current understanding of the cardiovascular risk associated with prediabetes.


Nothing to Disclose: IO, NU, HM, SE, JW, SD

27617 27.0000 FRI 679 A Endothelial Function and Prediabetes Status in African American and Caucasian Offspring of Parents with Type 2 Diabetes 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Aytekin Oguz*1, Gunes Alkaya2, Ozge Telci Caklili2 and Irem Bozkurt2
1Istanbul Medeniyet University Göztepe Training and Research Hospital, 2Istanbul Medeniyet University Goztepe Training and Research Hospital


Aspirin use for secondary prevention provides significant and clinically important reductions in the risk of  cardiovascular events. Recommendations for using aspirin in primary prevention has changed substantially in the past  years. American Diabetes Association (ADA) recommends (2015) considering aspirin therapy as a primary prevention strategy in patients with diabetes with increased cardiovascular risk (10-year risk >10%). Major risk factors which increase the risk are age, family history of cardiovascular disease (CVD), hypertension, smoking, dyslipidemia, or albuminuria.

We evaluated real world aspirin use in diabetic patients admitted to diabetes outpatient clinics of Istanbul Medeniyet University Goztepe Training and Research Hospital. Six hundred seventy four ( f: 279, m: 395 ) diabetic patients were enrolled in the study group. Patients were grouped with their cardiovascular risk status for aspirin indication as defined by ADA 2015 recommendations. Aspirin users for an indication other then antiagregant purpose were excluded.

Among 322 patients (M:180, F: 142) who have a net indication of aspirin use according to ADA recommendations (age>50 y for males, and>60 y for females plus at least one additional risk factor) 83 of men (45.1%), and 70 of women (49.3%) were not using aspirin or any other antiagregant medication. There were 50 patients (M:18, F:32) in whom aspirin use is not indicated for any other disease or their risk group as a diabetic (age<50 y for males, and<60 y for females and no additional major risk factor), 2 of women(11.1%), and 4 of men (12.5%)  were using aspirin. There were 143 patients with known CVD and  27 of them (18,9 %) were not using aspirin or any other antiagregant.

We found remarkable underuse of aspirin in our diabetes patients. Also there are considerable amount of patients who use  aspirin although not indicated.


Nothing to Disclose: AO, GA, OT, IB

27745 28.0000 FRI 680 A Aspirin Underuse in Patients with Diabetes Mellitus: Real World Data from Turkey 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Megan Beauchemin*, Anisha Contractor, Ivy Bergquist, Amy Davidoff and Karen L Houseknecht
University of New England, Biddeford, ME


Atypical  antipsychotic drugs (AA), including risperidone (RIS), are FDA approved for the treatment of mood disorders including schizophrenia and bipolar disorder and are increasingly prescribed off-label for diverse  indications including attention deficit hyperactivity disorder (ADHD/ADD), insomnia, post-traumatic stress disorder (PTSD) and aggression (autism and dementia).  AA medications cause significant endocrine and metabolic side effects including obesity, insulin resistance, hyperglycemia, hyperprolactinemia and dyslipidemia. Clinical studies have shown cardiovascular morbidities such as ventricular arrhythmias and sudden cardiac deaths linked with AA therapy, however the mechanisms underlying adverse endocrine, metabolic and cardiac outcomes are poorly understood.  AA have complex pharmacology, antagonizing several G-protein coupled receptors including: dopamine, serotonin, adrenergic and muscarinic receptors. MicroRNAs (miRNAs) are short noncoding RNAs that serve as powerful post-transcriptional regulators of gene expression, are associated with cardiac injury and disease, and have been recently shown to be aberrantly expressed in the plasma of patients treated with AA.  We hypothesize that antipsychotic drugs may “prime” the heart for metabolic disease via direct regulation of cardiac miRNAs.     To determine whether miRNAs are acutely regulated in the heart following AA administration, male Sprague-Dawley rats were treated orally with clinically relevant doses of RIS (0.03 or 0.3 mg/kg) or vehicle (V; methyl cellulose).  Plasma and apical sections of the ventricle were collected at 2, 6 and 24 hours post-dose.  As expected, prolactin in plasma was increased by 249% 2 hours post-dose vs. V (P<0.005).  Expression of a panel of miRNAs was also evaluated by qRT-PCR, including miR-21, miR-133a, miR-365, miR-101, miR-132 and miR-146. Of these, miR-133a, miR-21 and miR-365 showed dynamic regulation in response to RIS treatment. miR-133a was upregulated in the heart by ~30% and ~55%  at 2 hours and 6 hours following RIS treatment, respectively, compared to V (P<0.01), before returning to levels comparable to vehicle by 24 hr.  miR-365 also showed an early response to RIS exposure, being elevated by 2 hr  post dose (P<0.05).  Interestingly, miR-365 was downregulated in the hearts of RIS-treated rats by 24 hr, compared to V (P=0.057). miR-21 showed elevated expression of ~35% in the heart by 6 hours post RIS treatment compared to V (P<0.05) before returning to baseline levels by 24 hr.    Our data demonstrate an acute miRNA response in the heart following a single dose of RIS.  We are currently investigating the early regulation of these miRNAs, which are known to be associated with cardiac hypertrophy, diabetic cardiomyopathy, fibrosis, and heart failure, as potential mechanisms linking cardiometabolic disease and adverse cardiac events with AA medications.


Nothing to Disclose: MB, AC, IB, AD, KLH

25939 29.0000 FRI 681 A Acute Treatment with the Antipsychotic Drug, Risperidone, Regulates microRNAs Associated with Cardiovascular Disease 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Alessandra Bitto*1, Carmelo Gabriele Pizzino1, Giovanni Pallio1, Federica Galfo2, Natasha Irrera1, Domenica Altavilla2 and Francesco Squadrito1
1University of Messina, Messina, Italy, 2University of Messina


Atherosclerosis is mainly an inflammatory disease and a role for COX-1/2 and 5-lipoxygenase has been proposed in the last decade. Two naturally derived inhibitors of these enzymes, baicalin and catechin, have been combined in a proprietary blend, named flavocoxid. Flavocoxid has been proven effective in reducing the inflammatory cascade and ameliorating the clinical features of several inflammatory diseases. In addition this formula is already FDA approved as a medical food.

We tested the hypothesis that flavonoids contained in flavocoxid may have beneficial effects in the well known ApoE knock out atherosclerosis mouse model. Animals were 4-5 weeks old at the beginning of the study and were randomized to receive normal diet or high fat diet for 14 weeks. Concomitantly both groups were randomly assigned to receive flavocoxid (at the human equivalent dose of 500 mg/day) or placebo by oral gavage for the whole study period.

Every 2 weeks we assessed food intake, triglycerides and cholesterol blood levels. At the end of the 14 weeks animals were sacrificed and the aorta, liver, and blood were collected for further analysis. The administration of flavocoxid resulted in a reduced deposition of fat in the atherosclerotic lesions, with decreased intima-media thickness. In the liver an increased PPAR-alpha expression was observed together with increased expression of the AMPK-alpha in animals that received flavocoxid. Finally lipid profile was also improved by the flavonoids’ blend. 

These pre-clinical results suggest a potential use of flavocoxid, in the very next future, to halt the progression of the atherosclerotic disease.


Nothing to Disclose: AB, CGP, GP, FG, NI, DA, FS

27261 30.0000 FRI 682 A Flavonoids Contained in Flavocoxid Reduce the Extent of Atherosclerotic Lesions in the ApoE KO Mouse 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 651-682 7692 1:15:00 PM Cardiometabolic Risk/Vascular Biology (posters) Poster

Farid Saad*1, Aksam A Yassin2, Ahmad Haider3, Karim Sultan Haider3, Gheorghe Doros4 and Abdulmaged M Traish5
1Bayer Pharma AG, Berlin, Germany, 2Segeberger Kliniken, Norderstedt, Germany, 3Private Urology Practice, Bremerhaven, Germany, 4Boston University School of Public Health, Boston, MA, 5Boston University School of Medicine, Boston, MA


Background:While short-term studies using testosterone therapy (TTh) in hypogonadal men with type 2 diabetes mellitus (T2DM) have yielded inconsistent results, long-term treatment has shown beneficial effects of TTh. There is no information, however, whether TRT has benefits in hypogonadal men with prediabetes.

Material and methods: Men presenting to urologists with various complaints were screened for hypogonadism and, if found hypogonadal, offered TTh. Those who had received at least 1 year of treatment with testosterone undecanoate 1,000 mg injections (TU) were entered into two independent, prospective, observational, cumulative registry studies. 109 men with prediabetes, defined as baseline HbA1cfrom 5.7 to 6.4%, were analysed. TU was administered  in 3-month intervals following an initial 6-week interval for up to 8 years. At each or each other visit, anthropometric and metabolic parameters were measured. Patients whose TTh was temporarily interrupted were excluded from the analysis.

Results: Mean age was 57.37±8.99 years. Mean weight decreased from 96.15±13.05 to 84.14±6.98 kg by -14.58±0.68 kg, percent change from baseline -14±0.65%. Waist circumference decreased from 103.8±6.88 to 94.32±4.53 cm by -9.62±0.44 cm. BMI decreased from 30.55±4.35 to 27.04±2.55 kg/m2 by -4.66±0.23 kg/m2. Waist-to-height ratio decreased from 0.58±0.04 to 0.53±0.03. All anthropometric measures were statistically significant vs. baseline (p<0.0001) and improved progressively with statistical significance compared to the previous year for 6 to 7 years.

Fasting glucose decreased from 5.43±0.68 to 4.63±0.67 mmol/L (p<0.0001) by -0.94±0.11 mmol/L reaching a plateau after 1 year. HbA1cdecreased from 5.9±0.21 to 5.38±0.26% (p<0.0001) by -0.59±0.04% with statistical significance compared to the previous year for the first 3 years.

The triglyceride:HDL ratio, a surrogate parameter of insulin resistance, declined from 5.62±2.61 to 2.6±0.74 (p<0.0001). The product of fasting glucose and triglycerides (TyG Index), another surrogate for insulin resistance, improved from 4.04±0.17 to 3.81±0.14.

No patient progressed from prediabetes to T2DM. All but 4 patients’ last measured HbA1cwas <5.7%.

Lipids, blood pressure, liver transaminases and C-reactive protein all improved significantly.

3 patients dropped out, 2 due to relocation, 1 lost to follow-up. There were no major adverse cardiovascular events during the full observation time.

Conclusion: Hypogonadal men with prediabetes showed clinically meaningful and sustainable weight loss as well as improvements in glycaemic control when receiving long-term treatment with testosterone. No patient advanced from prediabetes to overt T2DM. TRT seems to be effective to improve anthropometric and metabolic parameters in hypogonadal men and in preventing progression from prediabetes to T2DM, thereby potentially reducing cardiometabolic risk.


Disclosure: FS: Employee, Bayer Schering Pharma. AAY: Investigator, Bayer Schering Pharma, Speaker, Bayer Schering Pharma. AH: Investigator, Bayer Schering Pharma. GD: Coinvestigator, Bayer Schering Pharma. Nothing to Disclose: KSH, AMT

25585 3.0000 FRI 685 A Progression from Prediabetes to Diabetes Type 2 Is Prevented in 109 Hypogonadal Men Treated with Testosterone for up to 8 Years 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 683-707 7707 1:15:00 PM Novel Treatment for Diabetes (posters) Poster

Aksam A Yassin*1, Ahmad Haider2, Gheorghe Doros3, Abdulmaged M Traish4 and Farid Saad5
1Segeberger Kliniken, Norderstedt, Germany, 2Private Urology Practice, Bremerhaven, Germany, 3Boston University School of Public Health, Boston, MA, 4Boston University School of Medicine, Boston, MA, 5Bayer Pharma AG, Berlin, Germany



While hypogonadism is highly prevalent in men with T2DM, effects of testosterone therapy (TTh), especially in short-term studies, is still controversial. This study investigated effects of long-term TRT in hypogonadal men with T2DM.


Hypogonadal men receiving TTh with testosterone undecanoate injections 1000 mg (TU) every 3 months following an initial 6-week interval in urological offices were entered into 2 registries. A pooled subgroup analysis was done in all 161 men with T2DM who received continuous TTh up to 8 years. T2DM was being treated by the respective family physicians. At each or each other visit, anthropometric and metabolic parameters were measured.


Mean baseline age was 60.67±5.62 years.

Mean baseline HbA1c was 7.97±0.86%. 22 patients (13.7%) were within an HbA1c target of 7.0%, 6 within an HbA1c target of 6.5%. Under TTh, HbA1c decreased from 7.97±0.86 to 7.46±0.77% after 1 year, 7.02±0.7% after 2 years, 6.75±0.67% after 3 years, 6.54±0.61% after 4 years, 6.31±0.55% after 5 years, 6.16±0.55% after 6 years, 6.0±0.51% after 7 years, and 5.76±0.52% after 8 years. These changes were statistically significant vs. baseline at each year (p<0.0001) and also statistically significant vs. previous year throughout the observation time. The mean change from baseline was -2.13±0.05%.

At the last observation, 150 patients (93.2%) were within an HbA1c target of 7.0%, 124 (77.0%) patients within an HbA1c target of 6.5%.

Fasting glucose decreased from 6.61±1.51 to 5.07±0.47 mmol/L (p<0.0001), mean change from baseline -1.63±0.10 mmol/L, reaching a plateau after 2 years.

2.5% of men had normal weight, 11.8% were overweight, and 85.7% obese. Mean weight decreased from 109.37±14.3 to 89.03±9.3 kg, change from baseline -21.13±0.53 kg, percent change from baseline -18.67±0.44%. Waist circumference decreased from 109.8±8.46 to 98.15±6.73 cm, change from baseline -12.37±0.29 cm. BMI decreased from 35.13±4.58 to 28.86±2.95 kg/m2, change from baseline -6.81±0.18 kg/m2. Waist-to-height ratio decreased from 0.62±0.05 to 0.56±0.04. All anthropometric measures were statistically significant vs. baseline (p<0.0001) and improved progressively with statistical significance compared to the previous year.

Lipid pattern, blood pressure, liver transaminases and C-reactive protein improved significantly.

No patient dropped out. Medication adherence was 100% as injections were administered in the office. There were no major adverse cardiovascular events during the full observation time.


Hypogonadal men with T2DM showed clinically meaningful and sustainable improvements in glycaemic control upon long-term TTh. Major weight loss has probably contributed largely to these effects. TTh seems to effectively improve T2DM-related parameters in hypogonadal men, in addition to standard therapy, thereby potentially reducing cardiometabolic risk.


Disclosure: AAY: Investigator, Bayer Schering Pharma, Speaker, Bayer Schering Pharma. AH: Investigator, Bayer Schering Pharma. GD: Coinvestigator, Bayer Schering Pharma. FS: Employee, Bayer Schering Pharma. Nothing to Disclose: AMT

26404 6.0000 FRI 689 A the Majority of Hypogonadal Men with Type 2 Diabetes Mellitus (T2DM) Achieve HbA1c Targets When Treated with Testosterone for up to 8 Years 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 683-707 7707 1:15:00 PM Novel Treatment for Diabetes (posters) Poster

Deepashree Gupta*1 and Stewart Gary Albert2
1Saint Louis University, St Louis, MO, 2St Louis Univ Sch of Med, Saint Louis, MO


Objective – An insulin infusion protocol (IIP) was instituted in medical and surgical ICUs for post-cardiac surgery stress hyperglycemia (SH), diabetes hyperglycemia (DH), and DKA. Prior to 2014, Saint Louis University Hospital (SLUH) used a proportionate delivery protocol which required hourly Insulin Infusin Rate (IIR) recalculation based on BG change.  A separate protocol was followed for diabetic ketoacidosis (DKA). A unified ICU protocol was designed in 2013 and implemented in 2014 for treatment of DH, DKA, and post-cardiothoracic surgery SH to meet SCIP criteria.  The protocol utilized conventional diabetes management techniques, i.e. 1) body weight based, 2) designed with the concept of basal plus correction factor,   3) pre-calculated insulin doses , 4) maximum IIR at higher BG and 6) progressively decreasing IIR as BG approaches designated target range (DTR) to limit hypoglycemia

Method - The IIP was assessed during one month for all patients in ICUs. The IIP was developed in Excel and our project is IRB exempt. The IIR is higher at BG >160mg/dL (0.5units/ kg/24 hour, defined as “Phase 1” to lower hyperglycemia), and decreases progressively as BG decline. There is an abrupt decrease in the IIR at a threshold BG of 160mg/dL, (rate 0.3units/kg/ 24hours defined as “Phase 2”, maintenance rate) to sustain BG between a DTR, 120 mg/dL-180 mg/dL. An acceptable target range (ATR) is considered between 100mg/dL-200mg/dL. A correction factor for BG >120mg/dL is added to the IIR by estimating Total Daily Dose (TDD) of insulin.  IIR is pre-calculated and stored in the electronic medical record system (EPIC) in weight based columns from 40kg through 150kg with increments of 10 kg.  Nurses titrate IIR hourly based only on the current BG. When BG has been in target, the frequency of BG determinations is decreased to every 2 hours and then to every 4 hours.  The protocol is designed to accommodate intravenous glucose but not to maintain glycemic control once the patient starts eating, at which point is replaced by a subcutaneous insulin regimen. When the protocol is utilized for DKA, all other non-insulin aspects of DKA protocol are maintained 

Research design- A convenience sample of 62 patients (20 SH, 39 DH, 3 DKA) were assessed. 

Results- IIP maintained BG 144.5 ± 24.7mg/dL for SH within an acceptable target range (ATR) (100mg/dL-200mg/dL) 95% of the time.  There were no BG > 240mg/dL or <70 mg/dL.  For DH, initial BG was 262 ± 88 mg/dL, and IIP maintained BG 165 ± 40 mg/dL within an ATR (81%) of time.  The time within ATR differed with DH alone (91%), on vasopressors (61%) or on glucocorticoids (84%), p<0.0001.

Conclusion- This IIP achieves BG targets of 100-200 mg/dL in majority of ICU populations. There were few episodes of hypoglycemia with BG <70mg/dL and none less than 40 mg/dL. The protocol adapts to increased insulin requirements on vasopressors and steroids. The protocol has been accepted for use in all medical and surgical ICUs


Nothing to Disclose: DG, SGA

24353 7.0000 FRI 690 A A Unified Hyperglycemia and Diabetic Ketoacidosis (DKA) Insulin Infusion Protocol Based on an Excel Algorithm 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 683-707 7707 1:15:00 PM Novel Treatment for Diabetes (posters) Poster

Cathy Sun*, Bertha Wong, Janine Charisse Malcolm and Mary-Anne Doyle
University of Ottawa, Ottawa, ON, Canada


Background: Poor glycemic control in patients with type 1 diabetes (T1DM) is associated with increased risk of acute and chronic complications.  Attaining optimal glycemic targets is known to improve outcomes however self-management is often challenging and complex.  Smart-phone mobile applications (mobile apps) have been shown to be beneficial in the management of many chronic diseases. Despite a wide variety of mobile apps designed to assist with diabetes management, the benefits of these interventions in TIDM has not been established.

Objectives: 1) To conduct a systematic review of the literature evaluating the use of stand-alone mobile apps and mobile apps with text-messaging in the management of T1DM. 2) To identify and characterize apps applicable to the management of T1DM and available through Google play and Apple app stores.  

Methods: Medline and Embase databases were systematically searched to identity all observational and intervention studies published before December 2014. We additionally reviewed top-rated apps and summarized each app’s key features, reviews, and costs.  

Results: Our initial search yielded 1727 studies. Eight studies met inclusion and exclusion criteria.  Four of these studies focused on the role of stand-alone mobile apps in management of T1DM.  These studies were small (a total of 265 T1DM participants) and short in duration (mean follow-up 3-6 months).  None of these studies demonstrated a significant improvement in HbA1c.  However, 2 studies did demonstrate improved adherence with glucose monitoring: frequency of daily blood sugar checks increased from 3.29 to 3.57 (p<0.05), and 2.4 to 3.6 (p<0.05).  Four other studies investigated the role of mobile apps with a text messaging system.   These studies were equally small (total of 252 participants) but slightly longer duration (3-9 months follow-up).  There was a trend towards improved HbA1c (-0.33 to -1.28%) and one study demonstrated a significant reduction in the incidence of severe hypoglycemic events (IR 0.33; 95% CI 0.17-0.63 (app+text) vs 2.29; 95% CI 1.80-2.91(control).

A search of the Google Play and the Apple app stores revealed over 600 mobile apps that were potentially useful in the management of type 1 diabetes.  Thirty of these were highly rated.  Many featured the ability to log diabetes parameters, provided graphical analysis, and enabled the setup of reminders.

Conclusion: Despite a vast array of relatively inexpensive mobile apps and evidence to support the benefit of these tools in other chronic diseases, there is a paucity of well-designed studies evaluating the role of these tools in management of T1DM.  This study highlights the need for larger and longer studies to explore the efficacy of mobile apps in T1DM in optimizing outcomes, the subgroup populations that would benefit most from these tools, and the resources needed to support mobile-apps plus text-messaging systems.


Nothing to Disclose: CS, BW, JCM, MAD

26592 8.0000 FRI 691 A Improving Glycemic Control in Type 1 Diabetes with the Use of Smart-Phone Based Mobile Applications: A Systematic Review 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 683-707 7707 1:15:00 PM Novel Treatment for Diabetes (posters) Poster

Om Jitendra Lakhani* and Surender Kumar
Sir Ganga Ram Hospital, New Delhi, India



New onset of diabetes after transplant (NODAT) is defined by international consensus guidelines published in 2003.(1) The objective of this study was to compare a novel algorithm developed to negate the effects of tacrolimus and prednisolone in hospitalized patients with NODAT vis a vis standard basal-bolus insulin algorithm.


This was a prospective, randomized, open labelled pilot study. Institutional ethics committee permission was sought for the trial and was granted. The study adhered to the requirements of the Declaration of Helsinki. Written and informed consent was obtained from the study participants

Hospitalized patients with NODAT were randomized to either of the two groups- Group A or Group B. Patients in group A were to receive insulin according to the novel algorithm, while patients in group B received insulin as per standard basal-bolus insulin algorithm followed in our hospital. The novel algorithm included insulin NPH given along with prednisolone in appropriate dose to negate the effect of later and insulin Glargine given once a day to negate the effect of tacrolimus. The blood glucose readings in the two groups were compared using glucometrics. The P value of <0.05 was considered significant.


A total of 28 patients with NODAT were recruited for the study. The baseline characteristics of the two groups were matched. There were 8 case of liver transplant and 6 cases of renal transplant in each group.

The patients treated with novel algorithm (group A) had significantly lower prelunch (p=0.003) and overall premeal blood glucose (p = 0.025). The hypoglycemia events were low in both the groups with a median value close to zero. Patients treated with novel algorithm had significantly less hyperglycemic excursion events (defined as blood glucose >300 mg/dl). Patients treated with standard basal-bolus algorithm had higher glycemic variability compared to those treated with novel algorithm (p=0.006).


We developed a novel algorithm that was built to negate the effects of prednisolone and tacrolimus in patients with NODAT. Patients treated with the novel algorithm had lower prelunch blood glucose, less episodes of hyperglycemic excursions and had lower glycemic variability.


Nothing to Disclose: OJL, SK

23876 9.0000 FRI 692 A Novel Insulin Algorithm for Management of Hyperglycemia in Hospitalized Patients with New Onset of Diabetes after Transplant (NODAT): A Pilot Study 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 683-707 7707 1:15:00 PM Novel Treatment for Diabetes (posters) Poster

Jacques Beltrand1, Kanetee Busiah1, Cecile Godot1, Raphael Scharfmann2, jean-Marc Treluyer3, Caroline Elie4 and Michel Polak*5
1Hôpital Necker Enfants Malades, Assistance Publique Hôpitaux de Paris, Faculté de médecine Paris Descartes, Paris, France, 2INSERM, Paris, France, 3Hôpital Necker Enfants Malades, Assistance Publique Hôpitaux de Paris, Faculté de médecine Paris Descartes, paris, France, 4Hôpital Necker Enfants Malades, Assistance Publique Hôpitaux de Paris, Faculté de médecine Paris Descartes, Paris, 5Hôpital Universitaire Necker-Enfants Malades, Paris, France


Background: Sulfonylurea therapy allows a better metabolic control than insulin in patients with neonatal diabetes secondary to mutation in potassium channel. Its galenic form (tablets) is not suitable for children, as the dosage can't be easily modulated and as it induces large PK variations when administer to young children.

Objective:To measure relative biodisponibility of a new galenic form of glibenclamide and to assess its safety and tolerability.

Method:Open-label, cross over randomised phase 1 study in 18 healthy male subjects. Single oral administration, in fasted conditions of two new oral glibenclamide suspensions (0.83 mL of a 6 mg/mL suspension (S6), 8.33 mL of a 0.6 mg/mL suspension (S0.6)) and of 5mg of Daonilâ crushed tablet (DCT).

Results:When suspensions were administered, glibenclamide plasma concentrations peaked 0.5 hour earlier than observed with a DCT (median value of 2.5 hours post-dose versus 3.00 hours post-dose). Mean plasma peak Cmax values were similar for the two suspensions (S6: 201.71±71.43 ng/mL S0.6: 206.93±67.33 ng/mL, approximately 40% higher than the DCT one (148.34±46.74 ng/mL). Exposures were similar for the two suspension dosages (AUC0-¥ values:  S6: 1120.9 ±400.5 ng.h/mL, S0.6: 1172.3±422.0 ng.h/mL), and superior to that observed after DCT administration. Relative bioavailability was 121.6% for the 0.6 mg/mL and 114.1% for the 6 mg/mL formulations when compared to the DCT. Elimination half-lives were similar for the two suspensions (close to 8 hours) and a little shorter than that observed with DCT (10.45 hours). No adverse events were reported.

Conclusion: Suspension of glibenclamide appears to be more suitable for use in pediatric patients as its dosage can be adjusted to patients needs with great precision more easily. PK studies reported it to be better absorbed than glibenclamide tablets. Tolerance and acceptability are being evaluated in patients with neonatal diabetes (ClinicalTrials.gov Identifier: NCT02375828). Preliminary results shows that the solution doesn't induce a higher rate of hypoglycemia or a significant change in HbA1C in patients with neonatal diabetes


Nothing to Disclose: JB, KB, CG, RS, JMT, CE, MP

24902 10.0000 FRI 693 A Pharmacokinetic of a New Suspension of Glibenclamide for Use in Young Patients and Infants with Neonatal Diabetes 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 683-707 7707 1:15:00 PM Novel Treatment for Diabetes (posters) Poster

Suzanne Erinn Kingery*, Sarah Spurling, Michael B. Foster, Adetokunbo Omoruyi, Sara E Watson and Kupper Anthony Wintergerst
University of Louisville, Louisville, KY


Children and adolescents with type 1 diabetes often struggle with the psychological, social, and physical burdens of disease management.  Wendy Novak Diabetes Care Center at the University of Louisville developed a specialized, multi-disciplinary clinic to assist patients with type 1 diabetes who have difficulty maintaining good diabetes control. The Diabetes Adherence Resource Team (D.A.R.T.) is comprised of a pediatric endocrinologist, diabetes nurse practitioner, certified diabetes educator, dietician and a pediatric psychologist.  Patients  follow-up as often as every four to eight weeks, rather than the standard three-month follow-up. Criteria for inclusion in the D.A.R.T. clinic includes HbA1c greater than 12% for two or more consecutive visits, frequent emergency room visits due to diabetes, hospital admission for diabetic ketoacidosis (DKA) and/or significant psychological disorder and psychosocial stressors. Patients in the D.A.R.T. clinic can choose either individual or group visits.

Forty-three individuals, 23 male and 20 females, enrolled in the D.A.R.T. clinic. Average age at initial visit was 15.4 years (range 11-19 years). Effectiveness of this multi-disciplinary clinic was measured by reduction in HbA1c, increase in average daily blood glucose checks, reduction in ED utilization and decrease in DKA admissions.  Mean HbA1c at the visit prior to the initial D.A.R.T. visit was 12.4% but had already decreased to 11.7% at the initial D.A.R.T. visit (p<0.0001, n=43). Mean HbA1c further decreased to 10.9 % at the first follow-up D.A.R.T. visit compared to the initial visit,(p<0.0001, n=18).  Average daily blood glucose checks had already increased to 1.97 at initial D.A.R.T visit compared to 1.39 at the visit prior to enrolling in the D.A.R.T. clinic (p<0.0001, n=43). Increase in average daily blood glucose checks continued to the first follow-up visit at 2.16 compared to the initial visit (p<0.0001, n=18). There was no difference in HbA1c (p=0.57) or average daily glucose checks (p=0.16) from initial D.A.R.T. visit to the follow-up visit between the individual or group visit formats. Since enrolling in the clinic, two patients have been admitted in DKA, but none have utilized the emergency room.

D.A.R.T. clinic is a more comprehensive diabetes visit that has the potential to improve the medical and behavioral health outcomes of individuals with type 1 diabetes. While this clinic is new in concept to our institution, the results of this targeted group indicate a positive influence on overall health and well-being of high risk youth with type 1 diabetes. As this clinic becomes more established and patients are followed for longer periods, we will be able to determine the long-term efficacy of this type of multi-disciplinary team intervention.


Disclosure: KAW: Advisory Group Member, Tandem. Nothing to Disclose: SEK, SS, MBF, AO, SEW

24793 11.0000 FRI 694 A Multi-Disciplinary Team Approach Improves Glycemic Control and Health Behaviors in High Risk Youth with Type 1 Diabetes Mellitus 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 683-707 7707 1:15:00 PM Novel Treatment for Diabetes (posters) Poster

Keren Zhou*1, Mia Williams2, Mary Angelynne Esquivel3, Anne Song1, Farah Rahman4, James Bena1, Simon Lam1, Deborah Rathz1 and M. Cecilia Lansang1
1Cleveland Clinic Foundation, Cleveland, OH, 2University of California San Francisco, San Fancisco, CA, 3Brown University, Providence, RI, 4Case Western Reserve University, Cleveland, OH


Hyperglycemia is a common phenomenon in the hospital and is a significant risk factor for prolonged hospitalization and mortality. Despite inpatient diabetes management guidelines recommending an overlap of basal insulin before discontinuing intravenous insulin infusion (IVII), there is a paucity of studies on transitioning from IVII to subcutaneous insulin (SCI). We aimed to compare glucose control and insulin dosing between 3 groups of patients in the medical ICU (MICU) transitioning from IVII to SCI: 1) NB (did not receive basal insulin), 2) CB (given basal insulin at a correct time period of up to 12 hours prior to IVII discontinuation, and 3) IB (received basal insulin at an incorrect time of 13-24 hours prior to IVII discontinuation or after IVII discontinuation). 

We conducted a retrospective review of patients admitted to the Cleveland Clinic MICU from June 2013 to January 2014 who received IVII. Patient characteristics, blood glucose levels (BG), insulin doses, and insulin dose per body weight in kg were compared using ANOVA for continuous variables. Time points used were at IVII discontinuation and at 1, 4, 8, 12 and 24 hours after discontinuation. 

There were 280 patients (NB 171, CB 50, IB 59), 55% male, mean age 58 + 17 years, and 75% with a known diagnosis of diabetes (NB 63%, CB 94%, IB 95%). Sixty-one percent did not receive any basal insulin. NB had better BG compared to IB and CB at the 8, 12, and 24 hour time points. IB and CB achieved comparable BG control for all time points with mean values mostly above 200 mg/dL. NB received 10.1 + 9.2 units of rapid- or short-acting insulin in 24 hr. CB received a significantly higher amount of basal insulin compared to IB (absolute amount 26.6 + 20.8 vs 18 + 12.3 units, p=0.012; dose per body weight 0.31 + 0.21 vs 0.22 + 0.15 unit/kg, p=0.017). CB and IB received a comparable amount of rapid- or short-acting insulin over the first 24 hours (absolute 15.8 +12.4 vs 12 + 9.2 unit, p=0.058, and per body weight 0.23 + 0.19 vs 0.2 + 0.17 unit/kg, p=0.408). 

There was a high rate of non-overlap with basal insulin when IVII was converted to SCI (NB plus IB).  However, NB had a greater percentage of no known diabetes, received a lower dose of insulin, and had consistently better BG than CB and IB, implying that this group did not necessarily require basal insulin after IVII discontinuation. Neither CB nor IB patients achieved ideal glycemic control of BG <180 mg/dL, suggesting that even if basal insulin was given at the appropriate time, 0.3 unit/kg is inadequate to achieve glucose control.


Nothing to Disclose: KZ, MW, MA, AS, FR, JB, SL, DR, MCL

25332 12.0000 FRI 695 A Insulin Doses and Glycemic Control during Transition from Intravenous to Subcutaneous Insulin in the Medical ICU 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 683-707 7707 1:15:00 PM Novel Treatment for Diabetes (posters) Poster

Tiffany Schumaker*1, Marisa Censani1, Elizabeth Mauer2 and Zoltan Antal1
1Weill Cornell Medicine New York Presbyterian, New York, NY, 2Weill Cornell Medicine, New York, NY


Background: Metformin therapy is associated with a higher prevalence of biochemical B12 deficiency in adults with diabetes, likely secondary to its effects on intestinal absorption.  Few studies to date have investigated the effects of metformin on B12 levels in children and adolescents and no studies have investigated the effects on markers of anemia.  No consensus regarding screening for B12 deficiency exists for adults or children on metformin. The aim of the study was to evaluate the association of metformin therapy with vitamin B12 levels and anemia in children and adolescents.

Methods:  This was a prospective study conducted in the pediatric endocrine outpatient clinics at Weill Cornell Medical College.  Vitamin B12 level, hemoglobin, hematocrit, mean corpuscular volume (MCV) and anthropometric parameters were measured in metformin treated subjects, between 8 and 18 years of age, prior to metformin therapy initiation and at 2 follow up visits (3 and 6 months) post initiation.    

Results:  Twenty- four subjects were recruited for the study (70% female, 30% male; average age 14.2 +/- 2.24 years); 16 patients had 3 month data and 14 patients had 6 month data available for analysis.  The mean BMI percentile was 96.1% and BMI z-score was 2.1 and the majority had evidence of insulin resistance (mean HOMA-IR 4.97+/-3.85, mean fasting insulin 22.45+/- 15.79 uIU/ml).  Forty two percent of patients had pre-diabetes and one subject was diagnosed with type 2 diabetes mellitus.  The mean initial B12 level was 414.6+/- 177pg/ml.  There were no subjects with vitamin B12 deficiency (level <175pg/ml) at baseline.

Vitamin B12 concentration was noted to decrease post treatment start with mean change in vitamin B12 concentration of 5.4 pg/ml at 3 months (n=16) and -27.5pg/ml at 6 months (n=14)(p=0.85, 0.25, respectively).  One subject was found to be vitamin B12 deficient at 6 months (153pg/ml) and notably also had the lowest baseline B12 level (175pg/ml).  Among the subjects with both 3 and 6 month data (n=8), the mean change in B12 level was -19.13pg/ml from 0-3 months, -21.38pg/ml from 3-6 months and -40.5pg/ml from 0-6months.  (p= 0.67, 0.43, 0.28, respectively). There was a statistically significant increase in MCV between 3 and 6 months of metformin therapy of 1.04+/- 0.90fL (p= 0.022).  No statistically significant change was noted for hemoglobin or hematocrit at 3 or 6 months.

Conclusion:  Although change in vitamin B12 was not statistically different after metformin initiation, a declining trend was found.  MCV significantly increased between 3 and 6 months of metformin therapy, a change that may be the first sign of vitamin B12 deficiency in children treated with metformin.  Further monitoring of vitamin B12 levels along with markers of anemia may be warranted after metformin introduction, especially in those patients that have borderline B12 levels at initiation.

Nothing to Disclose: TS, MC, EM, ZA



Nothing to Disclose: TS, MC, EM, ZA

24830 13.0000 FRI 696 A Metformin Therapy Is Associated with Changes in Vitamin B12 Level and Mean Corpuscular Volume in Children and Adolescents 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 683-707 7707 1:15:00 PM Novel Treatment for Diabetes (posters) Poster

Eric Zijlstra*1, Marek Demissie2, Tina Graungaard2, Tim Heise1, Leszek Nosek1 and Bruce W. Bode3
1Profil, Neuss, Germany, 2Novo Nordisk A/S, Søborg, Denmark, 3Atlanta Diabetes Assoc, Atlanta, GA


Ultra-fast-acting insulins may be advantageous for patients on continuous subcutaneous insulin infusion (CSII), provided that such insulins are compatible and safe to use in this application. The primary objective of this two-center, randomized, double-blind, parallel-group trial was to evaluate CSII compatibility of faster-acting insulin aspart (faster aspart) and insulin aspart (IAsp) in 37 adults (mean [± SD] age, 44.3 ± 14.6 years) with type 1 diabetes (T1D; duration, 24.1 ± 12.4 years; HbA1c 7.5 ± 0.7%) using their own MiniMed Paradigm® pump with either Quick-Set® or Silhouette®infusion sets. After a 2-week run-in period with IAsp, subjects received either faster aspart (n=25) or IAsp (n=12) for 6 weeks. Basal rates of insulin infusion were optimized during the first week of treatment. In subsequent weeks, bolus insulin delivery parameters were optimized. Insulin compatibility was evaluated by the number of potential occlusions documented, prompted by indirect observations (e.g. pump alert, observed leakage or unexplained hyperglycemia). In addition, subjects performed macroscopic evaluations of the infusion set and reservoir after routine changes (every 72 h) and whenever an occlusion was suspected; laboratory microscopic and macroscopic examinations of the reservoir and infusion set were performed at each weekly site visit.

After 6 weeks of treatment, no microscopically confirmed episodes of infusion-set occlusion were observed in either treatment arm (of the 219 infusion sets evaluated). Seven possible infusion set occlusions were reported by five subjects in the faster aspart group (after [mean ± SD] 1.19 ± 0.91 days of use), compared with none in the IAsp group. None of the possible infusion-set occlusions were associated with an observed plug in the pump; all but one were prompted by hyperglycemic events and the remaining episode was prompted by leakage. Laboratory macroscopic and microscopic evaluation of the infusion sets was possible in three cases, but showed no color change or particle or crystal formation. Laboratory microscopic evaluation of the infusion sets detected minimal particles on two occasions with faster aspart, classified as ‘unlikely related to insulin’ (gray-shadowed particles in one case and suspected silicone particles in the other).

The estimated mean change in HbA1cfrom baseline to Week 6 favored faster aspart, but was not statistically significantly different from IAsp in this small-scale, short-term trial (estimated treatment difference [ETD]: –0.14% [95% CI: –0.40; 0.11]). Similar trends were observed for fructosamine (ETD: –11.3 µmol/l [–26.4; 3.8]). No safety issues were found.

In conclusion, faster aspart was compatible with CSII use, with no microscopically confirmed infusion-set occlusions, and demonstrated a trend towards improved glycemic control.


Disclosure: EZ: n/a, Novo Nordisk, n/a, Dance Biopharm, Inc. MD: Employee, Novo Nordisk, Stock/Shares owner, Novo Nordisk. TG: Employee, Novo Nordisk. TH: Researcher, Adocia, Researcher, Astra Zeneca, Researcher, BD, Researcher, Biocon, Researcher, Boehringer Ingelheim, Researcher, Dance Pharmaceuticals, Researcher, Grünenthal, Researcher, Eli Lilly, Researcher, Medtronic, Researcher, Novo Nordisk, Researcher, Novartis Pharmaceuticals, Researcher, Sanofi, Researcher, Senseonics, Speaker Bureau Member, Eli Lilly, Scientific Board Member, Mylan, Speaker Bureau Member, Novo Nordisk, travel grant, Eli Lilly, travel grant, Mylan, Travel grant, Novo Nordisk, Advisory Group Member, Novo Nordisk. BWB: Speaker Bureau Member, Novo Nordisk, Speaker Bureau Member, Merck & Co., Speaker Bureau Member, Medtronic Minimed, Speaker Bureau Member, GlaxoSmithKline, Speaker Bureau Member, Astra Zeneca, Researcher, Sanofi, Researcher, Pfizer, Inc., Speaker Bureau Member, Sanofi, Researcher, Novo Nordisk, Researcher, NIH, Researcher, Medtronic, Researcher, MannKind, Researcher, Boehringer Ingelheim, Researcher, Lilly USA, LLC, Researcher, Lexicon, Researcher, JDRF, Researcher, Jansen Pharmaceuticals, Researcher, GlaxoSmithKline, Researcher, DexCom, Researcher, Biodel, Researcher, BD, Researcher, Abbott, Employee, Atlanta Diabetes Associates, Consultant, Valeritas, Consultant, Sanofi, Consultant, Novo Nordisk, Consultant, Medtronic, Consultant, Jansen Pharmaceuticals, Consultant, BD, Consultant, Biodel, Consultant, Astra Zeneca, Speaker Bureau Member, Valeritas, Researcher, Aseko. Nothing to Disclose: LN

25681 14.0000 FRI 697 A Compatibility and Safety of Faster-Acting Insulin Aspart used in Continuous Subcutaneous Insulin Infusion Therapy in Patients with Type 1 Diabetes 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism Friday, April 1st 3:15:00 PM FRI 683-707 7707 1:15:00 PM Novel Treatment for Diabetes (posters) Poster

Mary M Janci*, Hilaire Thompson, Irl B Hirsch and Lorena Alarcon-Casas Wright
University of Washington Medical Center, Seattle, WA



Standards of diabetes care (DC) emphasize patients’ perspectives and advocate a patient-centered approach. Monitoring of blood glucose is a key self-management strategy to improve DC. Research on patients’ perspectives on the importance of DD, and satisfaction upon applying such data prospectively to control glucose is unknown. 


To assess patients’ perceptions on the importance of DD, frequency of discussion on strategies and shared decisions with diabetes providers (DP) and impact on future glucose control decisions.


Patients seen at the UWDCC, May to July 2014, on IPT and/or CGM participated in a voluntary/anonymous written survey. They were asked: 1) the importance of the DD service, 2) to rate (1 being the least, 5 being the most), the importance of different factors when discussing DD with DP: patterns of high/low glucose, effect of exercise, stress, alcohol, 3) whether they discussed solutions to deal with such variables with their DP, 4) whether they used the DD to make decisions about treating glucose, 5) their interest to learn advanced features of their devices after discussing DD, 6) whether, if their DP no longer provided DD service, would they find another DP who would, 7) DD at home, 8) to rate the importance of DD as part of the clinic visit. Patients were asked to self-report their A1C. Descriptive statistics (SPSS 19) were used for analysis. Pearson Chi-Square was used to compare groups by A1C: ≤6.5%, 7-7.5% and ≥ 8%.


Among 103 respondents on IPT and/or CGM, DD was considered an important service by 97% of them. 99 respondents reported A1C as follows:  ≤6.5% (n=24), 7-7.5% (n=53), ≥ 8% (n=22). The most frequently rated factor as highest importance for discussion was identification of high/low glucose patterns (85%), followed by the effect of exercise (47%), stress (35%), and alcohol (24%). Patients reported discussing solutions with DP for identified variables in 98% of cases. DD was reported useful to make prospective decisions on glucose control in 99% of patients. 85% reported interest in learning advanced pump features and 73% were self-downloading. DD as part of the clinic visit was ranked as a high to very high level of importance by 89% of patients. Notably, 60% would consider finding another DP should DD not be available. Responses among A1C groups did not differ, except that respondents with A1C ≤6.5% reported less interest in learning advanced pump features when compared to patients with higher A1C (p=0.04).    


In our Academic Institution, the ability to identify patterns of glucose control and use for care discussions using IPT and/or CGM DD was highly appreciated by patients among different degrees of A1C. Providing such service helps in efforts to mutually agree on goals and tailor individualized treatment decisions. This is facilitated by efficient and optimal use of technology to improve quality of care and clinical parameters.


Disclosure: IBH: Investigator, Novo Nordisk, Consultant, Abbott Laboratories, Consultant, Roche Pharmaceuticals. Nothing to Disclose: MMJ, HT, LACW

26355 15.0000 FRI 698 A Patient Perspectives on Personal Insulin Pump Therapy (IPT) and/or Continuous Glucose Monitoring Device (CGM) Downloaded Data (DD) in an Academic Clinical Setting 2016-04-04 Boston 2016-03-29 98th Annual Meeting of the Endocrine Society Diabetes & Glucose Metabolism