MiR-133 promotes cardiac reprogramming by directly repressing Snai1 and silencing fibroblast signatures

Naoto Muraoka, Hiroyuki Yamakawa, Kazutaka Miyamoto, Taketaro Sadahiro, Tomohiko Umei, Mari Isomi, Hanae Nakashima, Mizuha Akiyama, Rie Wada, Kohei Inagawa, Takahiko Nishiyama, Ruri Kaneda, Toru Fukuda, Shu Takeda, Shugo Tohyama, Hisayuki Hashimoto, Yoshifumi Kawamura, Naoki Goshima, Ryo Aeba, Hiroyuki YamagishiKeiichi Fukuda, Masaki Ieda

Research output: Contribution to journalArticle

135 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)1565-1581
Number of pages17
JournalEMBO Journal
Volume33
Issue number14
DOIs
Publication statusPublished - 2014 Jul 17

Fingerprint

Fibroblasts
Cardiac Myocytes
Epithelial-Mesenchymal Transition
MicroRNAs
Gene expression
Transcription Factors
Genes
Gene Expression

Keywords

  • cardiomyocyte
  • microRNA
  • reprogramming
  • Snai1
  • transcription factor

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)

Cite this

MiR-133 promotes cardiac reprogramming by directly repressing Snai1 and silencing fibroblast signatures. / Muraoka, Naoto; Yamakawa, Hiroyuki; Miyamoto, Kazutaka; Sadahiro, Taketaro; Umei, Tomohiko; Isomi, Mari; Nakashima, Hanae; Akiyama, Mizuha; Wada, Rie; Inagawa, Kohei; Nishiyama, Takahiko; Kaneda, Ruri; Fukuda, Toru; Takeda, Shu; Tohyama, Shugo; Hashimoto, Hisayuki; Kawamura, Yoshifumi; Goshima, Naoki; Aeba, Ryo; Yamagishi, Hiroyuki; Fukuda, Keiichi; Ieda, Masaki.

In: EMBO Journal, Vol. 33, No. 14, 17.07.2014, p. 1565-1581.

Research output: Contribution to journalArticle

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 2014, 'MiR-133 promotes cardiac reprogramming by directly repressing Snai1 and silencing fibroblast signatures', EMBO Journal, vol. 33, no. 14, pp. 1565-1581. https://doi.org/10.15252/embj.201387605
Muraoka, Naoto ; Yamakawa, Hiroyuki ; Miyamoto, Kazutaka ; Sadahiro, Taketaro ; Umei, Tomohiko ; Isomi, Mari ; Nakashima, Hanae ; Akiyama, Mizuha ; Wada, Rie ; Inagawa, Kohei ; Nishiyama, Takahiko ; Kaneda, Ruri ; Fukuda, Toru ; Takeda, Shu ; Tohyama, Shugo ; Hashimoto, Hisayuki ; Kawamura, Yoshifumi ; Goshima, Naoki ; Aeba, Ryo ; Yamagishi, Hiroyuki ; Fukuda, Keiichi ; Ieda, Masaki. / MiR-133 promotes cardiac reprogramming by directly repressing Snai1 and silencing fibroblast signatures. In: EMBO Journal. 2014 ; Vol. 33, No. 14. pp. 1565-1581.
@article{5511ae4eb49141688832d43e0a043618,
title = "MiR-133 promotes cardiac reprogramming by directly repressing Snai1 and silencing fibroblast signatures",
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.",
keywords = "cardiomyocyte, microRNA, reprogramming, Snai1, transcription factor",
author = "Naoto Muraoka and Hiroyuki Yamakawa and Kazutaka Miyamoto and Taketaro Sadahiro and Tomohiko Umei and Mari Isomi and Hanae Nakashima and Mizuha Akiyama and Rie Wada and Kohei Inagawa and Takahiko Nishiyama and Ruri Kaneda and Toru Fukuda and Shu Takeda and Shugo Tohyama and Hisayuki Hashimoto and Yoshifumi Kawamura and Naoki Goshima and Ryo Aeba and Hiroyuki Yamagishi and Keiichi Fukuda and Masaki Ieda",
year = "2014",
month = "7",
day = "17",
doi = "10.15252/embj.201387605",
language = "English",
volume = "33",
pages = "1565--1581",
journal = "EMBO Journal",
issn = "0261-4189",
publisher = "Nature Publishing Group",
number = "14",

}

TY - JOUR

T1 - MiR-133 promotes cardiac reprogramming by directly repressing Snai1 and silencing fibroblast signatures

AU - Muraoka, Naoto

AU - Yamakawa, Hiroyuki

AU - Miyamoto, Kazutaka

AU - Sadahiro, Taketaro

AU - Umei, Tomohiko

AU - Isomi, Mari

AU - Nakashima, Hanae

AU - Akiyama, Mizuha

AU - Wada, Rie

AU - Inagawa, Kohei

AU - Nishiyama, Takahiko

AU - Kaneda, Ruri

AU - Fukuda, Toru

AU - Takeda, Shu

AU - Tohyama, Shugo

AU - Hashimoto, Hisayuki

AU - Kawamura, Yoshifumi

AU - Goshima, Naoki

AU - Aeba, Ryo

AU - Yamagishi, Hiroyuki

AU - Fukuda, Keiichi

AU - Ieda, Masaki

PY - 2014/7/17

Y1 - 2014/7/17

N2 - 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.

AB - 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.

KW - cardiomyocyte

KW - microRNA

KW - reprogramming

KW - Snai1

KW - transcription factor

UR - http://www.scopus.com/inward/record.url?scp=84904580766&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84904580766&partnerID=8YFLogxK

U2 - 10.15252/embj.201387605

DO - 10.15252/embj.201387605

M3 - Article

C2 - 24920580

AN - SCOPUS:84904580766

VL - 33

SP - 1565

EP - 1581

JO - EMBO Journal

JF - EMBO Journal

SN - 0261-4189

IS - 14

ER -