CRISPR-Knockout Screen Identifies Dmap1 as a Regulator of Chemically Induced Reprogramming and Differentiation of Cardiac Progenitors

被引:12
作者
Yu, Jason S. L. [1 ,6 ]
Palano, Giorgia [2 ]
Lim, Cindy [3 ]
Moggio, Aldo [2 ]
Drowley, Lauren [3 ]
Plowright, Alleyn T. [4 ]
Bohlooly-Y, Mohammad [5 ]
Rosen, Barry S. [5 ]
Hansson, Emil M. [2 ]
Wang, Qing-Dong [3 ]
Yusa, Kosuke [1 ,7 ]
机构
[1] Wellcome Sanger Inst, Stem Cell Genet, Cambridge, England
[2] Karolinska Inst, KI AZ Integrated CardioMetab Ctr ICMC, Dept Med, Huddinge, Sweden
[3] AstraZeneca, Biosci Heart Failure Cardiovasc Renal & Metab, IMED Biotech Unit, Gothenburg, Sweden
[4] AstraZeneca, Med Chem Cardiovasc Renal & Metab, IMED Biotech Unit, Gothenburg, Sweden
[5] AstraZeneca, Discovery Sci, IMED Biotech Unit, Gothenburg, Sweden
[6] Francis Crick Inst, Dept Cell Biol, London, England
[7] Kyoto Univ, Stem Cell Genet, Inst Frontier Life & Med Sci, Kyoto 6068507, Japan
基金
英国惠康基金;
关键词
Cardiac progenitors; Chemical reprogramming; Clustered regularly interspaced short palindromic repeats-Cas9; Genome-wide screen; CpG methylation; SMOOTH-MUSCLE DIFFERENTIATION; STEM-CELLS; HEART REGENERATION; FIBROBLASTS; CARDIOMYOCYTES; CONTRIBUTE; HYPOXIA; RENEWAL;
D O I
10.1002/stem.3012
中图分类号
Q813 [细胞工程];
学科分类号
摘要
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
引用
收藏
页码:958 / 972
页数:15
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