NEUROD2 function is dispensable for human pancreatic β cell specification

Frontiers in Endocrinology

Endocrinology

Damla Taskin¹  Alireza Shahryari¹  Matthew Vanheusden¹  Aimée Bastidas-Ponce²  Mostafa Bakhti²  Michael Sterr²  Ingo Burtscher²  Changying Jing³  Perla Cota⁴  Lama Saber⁴  Heiko Lickert⁴ 

[1] Institute of Diabetes and Regeneration Research, Helmholtz Munich, Neuherberg, Germany;Institute of Diabetes and Regeneration Research, Helmholtz Munich, Neuherberg, Germany;German Center for Diabetes Research (DZD), Neuherberg, Germany;Institute of Diabetes and Regeneration Research, Helmholtz Munich, Neuherberg, Germany;German Center for Diabetes Research (DZD), Neuherberg, Germany;Munich Medical Research School (MMRS), Ludwig Maximilian University (LMU), Munich, Germany;Institute of Diabetes and Regeneration Research, Helmholtz Munich, Neuherberg, Germany;German Center for Diabetes Research (DZD), Neuherberg, Germany;School of Medicine, Technical University of Munich (TUM), Munich, Germany;

关键词: NEUROD2;    β cells;    endocrinogenesis;    iPSC differentiation;    endocrine cells;   

DOI  :  10.3389/fendo.2023.1286590

 received in 2023-08-31, accepted in 2023-10-09,  发布年份 2023

来源: Frontiers

【 摘 要 】

IntroductionThe molecular programs regulating human pancreatic endocrine cell induction and fate allocation are not well deciphered. Here, we investigated the spatiotemporal expression pattern and the function of the neurogenic differentiation factor 2 (NEUROD2) during human endocrinogenesis.MethodsUsing Crispr-Cas9 gene editing, we generated a reporter knock-in transcription factor (TF) knock-out human inducible pluripotent stem cell (iPSC) line in which the open reading frame of both NEUROD2 alleles are replaced by a nuclear histone 2B-Venus reporter (NEUROD2nVenus/nVenus). ResultsWe identified a transient expression of NEUROD2 mRNA and its nuclear Venus reporter activity at the stage of human endocrine progenitor formation in an iPSC differentiation model. This expression profile is similar to what was previously reported in mice, uncovering an evolutionarily conserved gene expression pattern of NEUROD2 during endocrinogenesis. In vitro differentiation of the generated homozygous NEUROD2nVenus/nVenus iPSC line towards human endocrine lineages uncovered no significant impact upon the loss of NEUROD2 on endocrine cell induction. Moreover, analysis of endocrine cell specification revealed no striking changes in the generation of insulin-producing b cells and glucagon-secreting a cells upon lack of NEUROD2.DiscussionOverall, our results suggest that NEUROD2 is expendable for human b cell formation in vitro.

【 授权许可】

Unknown   
Copyright © 2023 Cota, Saber, Taskin, Jing, Bastidas-Ponce, Vanheusden, Shahryari, Sterr, Burtscher, Bakhti and Lickert