High Dose Vitamin D Treatment Changes  T Helper Cells´ Gene Expression Profiles in Autoimmune Addison’s Disease

Presentation Number: OR07-2
Date of Presentation: April 2nd, 2017

Marissa Penna-Martinez1, Lennart Müggenburg2, Claudia Döring3, Dimitra Bogdanou1, Firouzeh Shoghi3, Martin-Leo Hansmann3 and Klaus Badenhoop*4
1University Hospital, Goethe-University Frankfurt am Main, Germany, 2University Hospital Frankfurt am Main, Frankfurt/Main, 3University Hospital Frankfurt am Main, Germany, 4Goethe-University Hospital, Frankfurt, Germany

Abstract

Introduction: Autoimmune Addison’s disease (AD) results from the immune-mediated destruction of the adrenal cortex requiring life-long gluco- and mineralocorticoid replacement. Most AD patients are vitamin D deficient.

Since vitamin D interacts with the glucocorticoid (GC) system and also regulates immune functions, the purpose of this pilot study was to evaluate the effect of three months cholecalciferol therapy on the gene expression in T helper cells of AD patients with hydrocortisone replacement. Methods: Nine AD patients on hydrocortisone (median dose: 20 mg/day) and a vitamin D deficiency (25(OH)D3 < 20 ng/ml) were included in a randomised, placebo-controlled study. During 3 months three patients received 4000 IU/d cholecalciferol and six patient’s placebo oil. At start (V1) and after 3 month (V3) 25(OH)D3 plasma concentration (radioimmunoassay) was measured and the gene expression profile of peripheral blood T helper cells was determined using microarray analysis (GeneChip Human 1.0 ST, Affymetrix). We evaluated the vitamin D treatment effects on the gene expression profiles by calculating their ratio between verum and placebo (expressed in fold changes=FC). Results: The 25(OH)D3 concentrations increased significantly after high dose VD (p < 0.05). In addition, in Th cells from AD patients, 67 annotated genes changed significantly after vitamin D treatment (21 down regulated, 46 upregulated genes). Of particular interest were 11 of 67 genes which possess transcription factor binding sites for the GC receptor (GR). Out of these, genes coding for the chemokine C-C motif ligand 3 (=CCL3 FC:-2.3;p=0.001), dual specificity phosphatase 1 (=DUSP1 FC:-1.6;p=0.04), jun oncogene (JUN FC: -2.6;p=0.02) and interleukin 1, beta (=IL1β FC:-1.8;p<0.03) were downregulated after vitamin D therapy. In contrast, genes which code for small nuclear RNA family (= SNORA38B FC:1.7;p<0.0004 and SNORA71D FC:1.6;p=0.01) and major histocompatibility complex (=HLADQA2; FC:2.4;p<0.02) showed a higher expression after vitamin treatment. Conclusion: In our pilot study, high dose VD treatment changes the gene expression profiles in T helper cells of AD patients resulting in the differential regulation of 67 genes (46 upregulated/21 downregulated). Since the transcription factor GR binds to the genes CCL3, DUSP1, JUN, IL1B, SNORA38B, SNORA71D and HLADQA2, we postulate that vitamin D exerts its actions on these genes in an indirect manner. Hereby ligand binding to the VDR leads to interaction with the transcription factor GR. Additionally, an indirect activation via activator protein 1 (AP-1) is also conceivable. To elucidate this mechanism further experiments are underway.

 

Nothing to Disclose: MP, LM, CD, DB, FS, MLH, KB