A Network of microRNAs Acts to Promote Cell Cycle Exit and Differentiation of Human Pancreatic Endocrine Cells.

Wen Jin,Jinzhao Wang,Allen Wang,Kelly A Frazer,Nicholas Vinckier,Ileana Matta,Yinghui Sui,Klaus H Kaestner,Bjoern Gaertner,Maike Sander,Andrea C Carrano,Kim-Vy Nguyen-Ngoc,Hung-Ping Shih,Francesca Mulas,Chun Zeng,Joshua Chiou

doi:10.1016/j.isci.2019.10.063

Abstract

SummaryPancreatic endocrine cell differentiation is orchestrated by the action of transcription factors that operate in a gene regulatory network to activate endocrine lineage genes and repress lineage-inappropriate genes. MicroRNAs (miRNAs) are important modulators of gene expression, yet their role in endocrine cell differentiation has not been systematically explored. Here we characterize miRNA-regulatory networks active in human endocrine cell differentiation by combining small RNA sequencing, miRNA over-expression, and network modeling approaches. Our analysis identified Let-7g, Let-7a, miR-200a, miR-127, and miR-375 as endocrine-enriched miRNAs that drive endocrine cell differentiation-associated gene expression changes. These miRNAs are predicted to target different transcription factors, which converge on genes involved in cell cycle regulation. When expressed in human embryonic stem cell-derived pancreatic progenitors, these miRNAs induce cell cycle exit and promote endocrine cell differentiation. Our study delineates the role of miRNAs in human endocrine cell differentiation and identifies miRNAs that could facilitate endocrine cell reprogramming.

Highlights

The potential to generate pancreatic beta cells from human pluripotent stem cells or via cell reprogramming from other cell sources holds promise for modeling causes of diabetes and cell replacement therapies (Benthuysen et al, 2016)
Identification of miRNAs Up-Regulated during Endocrine Cell Differentiation To identify miRNAs that are regulated during pancreatic beta cell differentiation, we conducted genome-wide small RNA sequencing in pancreatic progenitor cells derived from CyT49 human embryonic stem cells (Figure S1) and primary beta cells isolated from cadaveric human islets by fluorescence-activated cell sorting (Kameswaran et al, 2014) (Figure 1A)
By comparing expression levels of individual miRNAs in beta cells and pancreatic endoderm stage (PE) cells, we defined miRNAs induced during beta cell differentiation

Summary

Introduction

The potential to generate pancreatic beta cells from human pluripotent stem cells (hPSCs) or via cell reprogramming from other cell sources holds promise for modeling causes of diabetes and cell replacement therapies (Benthuysen et al, 2016). Studies in mice and zebrafish have demonstrated important roles for miRNAs in pancreatic endocrine cell development and beta cell function (Kaspi et al, 2014). Pancreatic progenitor cell-specific deletion of Dicer, an enzyme that is universally required for the functional maturation of miRNAs, results in reduced endocrine cell numbers (Lynn et al, 2007), whereas Dicer disruption in beta cells impairs insulin biogenesis (Melkman-Zehavi et al, 2011). At the level of individual miRNAs, miR-375 (Kloosterman et al, 2007; Poy et al, 2009) and miR-7 (Kredo-Russo et al, 2012; Latreille et al, 2014) have been identified as regulators of beta cell differentiation and function

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