Induction of human cardiomyocyte-like cells from fibroblasts by defined factors

Rie Wada, Naoto Muraoka, Kohei Inagawa, Hiroyuki Yamakawa, Kazutaka Miyamoto, Taketaro Sadahiro, Tomohiko Umei, Ruri Kaneda, Tomoyuki Suzuki, Kaichiro Kamiya, Shugo Tohyama, Shinsuke Yuasa, Kiyokazu Kokaji, Ryou Aeba, Ryohei Yozu, Hiroyuki Yamagishi, Toshio Kitamura, Keiichi Fukuda, Masaki Ieda

研究成果: Article査読

220 被引用数 (Scopus)

抄録

Heart disease remains a leading cause of death worldwide. Owing to the limited regenerative capacity of heart tissue, cardiac regenerative therapy has emerged as an attractive approach. Direct reprogramming of human cardiac fibroblasts (HCFs) into cardiomyocytes may hold great potential for this purpose. We reported previously that induced cardiomyocyte-like cells (iCMs) can be directly generated from mouse cardiac fibroblasts in vitro and vivo by transduction of three transcription factors: Gata4, Mef2c, and Tbx5, collectively termed GMT. In the present study, we sought to determine whether human fibroblasts also could be converted to iCMs by defined factors. Our initial finding that GMT was not sufficient for cardiac induction in HCFs prompted us to screen for additional factors to promote cardiac reprogramming by analyzing multiple cardiac-specific gene induction with quantitative RT-PCR. The addition of Mesp1 and Myocd to GMT up-regulated a broader spectrum of cardiac genes in HCFs more efficiently compared with GMT alone. The HCFs and human dermal fibroblasts transduced with GMT, Mesp1, and Myocd (GMTMM) changed the cell morphology from a spindle shape to a rod-like or polygonal shape, expressed multiple cardiac-specific proteins, increased a broad range of cardiac genes and concomitantly suppressed fibroblast genes, and exhibited spontaneous Ca2+ oscillations. Moreover, the cells matured to exhibit action potentials and contract synchronously in coculture with murine cardiomyocytes. A 5-ethynyl-2′-deoxyuridine assay revealed that the iCMs thus generated do not pass through a mitotic cell state. These findings demonstrate that human fibroblasts can be directly converted to iCMs by defined factors, which may facilitate future applications in regenerative medicine.

本文言語English
ページ(範囲)12667-12672
ページ数6
ジャーナルProceedings of the National Academy of Sciences of the United States of America
110
31
DOI
出版ステータスPublished - 2013 7月 30

ASJC Scopus subject areas

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