RRC ID 35802
Author Muraoka N, Yamakawa H, Miyamoto K, Sadahiro T, Umei T, Isomi M, Nakashima H, Akiyama M, Wada R, Inagawa K, Nishiyama T, Kaneda R, Fukuda T, Takeda S, Tohyama S, Hashimoto H, Kawamura Y, Goshima N, Aeba R, Yamagishi H, Fukuda K, Ieda M.
Title MiR-133 promotes cardiac reprogramming by directly repressing Snai1 and silencing fibroblast signatures.
Journal EMBO J
Abstract Fibroblasts can be directly reprogrammed into cardiomyocyte-like cells (iCMs) by overexpression of cardiac transcription factors or microRNAs. However, induction of functional cardiomyocytes is inefficient, and molecular mechanisms of direct reprogramming remain undefined. Here, we demonstrate that addition of miR-133a (miR-133) to Gata4, Mef2c, and Tbx5 (GMT) or GMT plus Mesp1 and Myocd improved cardiac reprogramming from mouse or human fibroblasts by directly repressing Snai1, a master regulator of epithelial-to-mesenchymal transition. MiR-133 overexpression with GMT generated sevenfold more beating iCMs from mouse embryonic fibroblasts and shortened the duration to induce beating cells from 30 to 10 days, compared to GMT alone. Snai1 knockdown suppressed fibroblast genes, upregulated cardiac gene expression, and induced more contracting iCMs with GMT transduction, recapitulating the effects of miR-133 overexpression. In contrast, overexpression of Snai1 in GMT/miR-133-transduced cells maintained fibroblast signatures and inhibited generation of beating iCMs. MiR-133-mediated Snai1 repression was also critical for cardiac reprogramming in adult mouse and human cardiac fibroblasts. Thus, silencing fibroblast signatures, mediated by miR-133/Snai1, is a key molecular roadblock during cardiac reprogramming.
Volume 33(14)
Pages 1565-81
Published 2014-7-17
DOI 10.15252/embj.201387605
PII embj.201387605
PMID 24920580
PMC PMC4198052
MeSH Analysis of Variance Animals Blotting, Western Cell Transdifferentiation / genetics Cell Transdifferentiation / physiology* Cloning, Molecular Fibroblasts / cytology Fibroblasts / metabolism* Flow Cytometry Gene Expression Regulation / physiology* Gene Knockdown Techniques Green Fluorescent Proteins Humans Immunohistochemistry Mice MicroRNAs / metabolism* Microarray Analysis Myocytes, Cardiac / cytology Myocytes, Cardiac / metabolism* Real-Time Polymerase Chain Reaction Snail Family Transcription Factors Transcription Factors / genetics* Transcription Factors / metabolism
IF 9.889
Times Cited 148
DNA material CSII-CMV-Venus (RDB05553) CSII-CMV-RfA (RDB04386) pCMV-VSV-G-RSV-Rev (RDB04393) pCAG-HIVgp (RDB04394).