The Tumor Suppressor p53 Regulates Induction of Functioning Anterior Pituitary Cells through Placodal Differentiation in Human Embryonic Stem Cells

Presentation Number: MON 370
Date of Presentation: April 3rd, 2017

Takashi Kono*1, Seiichirou Higuchi1, Sawako Suzuki1, Koutaro Yokote1 and Tomoaki Tanaka2
1Chiba University Graduate School of Medicine, Chiba, Japan, 2Chiba University Graduate School of Medicine, Chiba-city, Japan

Abstract

【Introduction】

Recently it is topic that the protocol of induction of functional pituitary cell using human embryonic stem cells (hESC) has been established. However, the molecular mechanism of differentiation induction process has not been clear enough. We focused on the transcription factor p53, which acts at the junction of tumor inhibitory signals and stem cell function control, and established the model of functional pituitary cell induction from hESCs in vitro, combination of CRISPR / Cas9, we examined the role of p53 in the differentiation process.

【Result】

We established placode not through embryonic body formation (early phase) and model of the T-pit / POMC, Pit1 / GH lineage functional endocrine cell (late phase) using hESCs. In the early phase expression of preplacode marker (SIX1, EYA1) was upregulated. In the late phase expression of T-pit or Pit-1 lineage factor was upregulated in a time-dependent and phase-specific manner, we confirmed the functionality and stimulus response by measurements at hormone in the culture solutions and immunostaining at day30. We established P53 knock-out hESCs and performed induction of placode and functioning pituitary cells using p53KO hESC, we observed the upregulation of SIX1, EYA1 in the early phase. Interestingly we confirmed observed the greater expression of TBX19 / NeuroD1 / POMC in late phase. From genomewide analysis using RNA-seq, a key molecule of the neural stem cell sorting PAX6 and the involvement of various non-coding RNA was suggested.

【Discussion】

P53 function in stem cells has been reported that the nuclear reprogramming suppression and three endoderm differentiation ability to control. We clarified that our results p53, as a new function of the p53 signaling, modulated the pituitary differentiation processes in vitro. To elucidate the difference between tumor suppression signal and the pituitary differentiation mechanism in p53, safe and efficient method for inducing application to development was considered to be expected.

 

Disclosure: TT: Investigator, ONO-Pharma. Nothing to Disclose: TK, SH, SS, KY