CRISPR-knockout screen identifies Dmap1 as a regulator of chemically induced reprogramming and differentiation of cardiac progenitors
File(s)
Published version
Author(s)
Type
Journal Article
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
Date Issued
2019-07-01
Date Acceptance
2019-03-02
Citation
Stem Cells, 2019, 37 (7), pp.958-972
ISSN
1066-5099
Publisher
Oxford University Press
Start Page
958
End Page
972
Journal / Book Title
Stem Cells
Volume
37
Issue
7
Copyright Statement
©2019 The Authors. STEM CELLS published by Wiley Periodicals, Inc. on behalf of AlphaMed Press 2019 This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
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Subjects
Biotechnology & Applied Microbiology
Cardiac progenitors
CARDIOMYOCYTES
Cell & Tissue Engineering
Cell Biology
Chemical reprogramming
Clustered regularly interspaced short palindromic repeats-Cas9
CONTRIBUTE
CpG methylation
FIBROBLASTS
Genome-wide screen
HEART REGENERATION
Hematology
HYPOXIA
Life Sciences & Biomedicine
Oncology
RENEWAL
Science & Technology
SMOOTH-MUSCLE DIFFERENTIATION
STEM-CELLS
Publication Status
Published
Date Publish Online
2019-04-23