MiR‐133 promotes cardiac reprogramming by directly repressing Snai1 and silencing fibroblast signatures
| Autor: | Shugo Tohyama, Keiichi Fukuda, Tomohiko Umei, Ryo Aeba, Shu Takeda, Rie Wada, Kohei Inagawa, Masaki Ieda, Kazutaka Miyamoto, Yoshifumi Kawamura, Ruri Kaneda, Hanae Nakashima, Naoto Muraoka, Mari Isomi, Mizuha Akiyama, Hisayuki Hashimoto, Hiroyuki Yamakawa, Naoki Goshima, Taketaro Sadahiro, Toru Fukuda, Hiroyuki Yamagishi, Takahiko Nishiyama |
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| Rok vydání: | 2014 |
| Předmět: |
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Western Green Fluorescent Proteins Biology Real-Time Polymerase Chain Reaction complex mixtures General Biochemistry Genetics and Molecular Biology Mice medicine Animals Humans Gene silencing Myocytes Cardiac MEF2C Cloning Molecular Fibroblast Molecular Biology Analysis of Variance Gene knockdown General Immunology and Microbiology GATA4 General Neuroscience Articles Fibroblasts Flow Cytometry Microarray Analysis Immunohistochemistry Embryonic stem cell MicroRNAs medicine.anatomical_structure Gene Expression Regulation Gene Knockdown Techniques Cell Transdifferentiation SNAI1 cardiovascular system Cancer research Snail Family Transcription Factors Reprogramming Transcription Factors |
| Zdroj: | The EMBO Journal. 33:1565-1581 |
| ISSN: | 1460-2075 0261-4189 |
| DOI: | 10.15252/embj.201387605 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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