CRISPR-Knockout Screen Identifies Dmap1 as a Regulator of Chemically Induced Reprogramming and Differentiation of Cardiac Progenitors. (23rd April 2019)

• Record Type: Journal Article
• Authors: Yu, Jason S. L.; Palano, Giorgia; Lim, Cindy; Moggio, Aldo; Drowley, Lauren; Plowright, Alleyn T.; Bohlooly-Y, Mohammad; Rosen, Barry S.; Hansson, Emil M.; Wang, Qing-Dong; Yusa, Kosuke
• Is Part Of: Stem cells. Volume 37:Number 7(2019)
• Page Start: 958
• Page End: 972
• Publication Date: 2019-04-23
• Subjects: Cardiac progenitors -- Chemical reprogramming -- Clustered regularly interspaced short palindromic repeats-Cas9 -- Genome-wide screen -- CpG methylation; Cloning -- Periodicals; Clone cells -- Periodicals; Stem cells -- Periodicals; Cell Differentiation -- Periodicals; Cell Division -- Periodicals; Clone Cells -- Periodicals; Hematopoietic Stem Cells -- Periodicals; Stem Cells -- Periodicals; 571.84
• DOI: 10.1002/stem.3012
• Languages: English
• ISSNs: 1066-5099

Abstract

Abstract : Direct in vivo reprogramming of cardiac fibroblasts into myocytes is an attractive therapeutic intervention in resolving myogenic deterioration. Current transgene-dependent approaches can restore cardiac function, but dependence on retroviral delivery and persistent retention of transgenic sequences are significant therapeutic hurdles. Chemical reprogramming has been established as a legitimate method to generate functional cell types, including those of the cardiac lineage. Here, we have extended this approach to generate progenitor cells that can differentiate into endothelial cells and cardiomyocytes using a single inhibitor protocol. Depletion of terminally differentiated cells and enrichment for proliferative cells result in a second expandable progenitor population that can robustly give rise to myofibroblasts and smooth muscle. Deployment of a genome-wide knockout screen with clustered regularly interspaced short palindromic repeats-guide RNA library to identify novel mediators that regulate the reprogramming revealed the involvement of DNA methyltransferase 1-associated protein 1 (Dmap1). Loss of Dmap1 reduced promoter methylation, increased the expression of Nkx2-5, and enhanced the retention of self-renewal, although further differentiation is inhibited because of the sustained expression of Cdh1. Our results hence establish Dmap1 as a modulator of cardiac reprogramming and myocytic induction. Stem Cells 2019;37:958–972 : Abstract : Chemically induced reprogramming using Alk5i coupled with hypoxia permits the conversion of cardiac fibroblast into cardiac progenitor, which can spontaneously give rise to cardiomyocyte and endothelial cell. Enrichment for proliferative cells leads to the derivation of chemically induced smooth muscle progenitor (ciSMP), which can give rise to myofibroblasts and smooth muscles. Modulation of Dmap1 enhances ciSMP derivation and self-renewal but compromises differentiation potency via modulation of Cdh1.