title: Induction of human cardiomyocyte-like cells from fibroblasts by defined factors.
abstract: 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 Ca(2+) 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.
authors.affiliation: Department of Clinical and Molecular Cardiovascular Research, School of Medicine, Keio University, Tokyo 160-8582, Japan.
authors.forename: Rie
authors.initials: R
authors.lastname: Wada
authors.forename: Naoto
authors.initials: N
authors.lastname: Muraoka
authors.forename: Kohei
authors.initials: K
authors.lastname: Inagawa
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Yamakawa
authors.forename: Kazutaka
authors.initials: K
authors.lastname: Miyamoto
authors.forename: Taketaro
authors.initials: T
authors.lastname: Sadahiro
authors.forename: Tomohiko
authors.initials: T
authors.lastname: Umei
authors.forename: Ruri
authors.initials: R
authors.lastname: Kaneda
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Suzuki
authors.forename: Kaichiro
authors.initials: K
authors.lastname: Kamiya
authors.forename: Shugo
authors.initials: S
authors.lastname: Tohyama
authors.forename: Shinsuke
authors.initials: S
authors.lastname: Yuasa
authors.forename: Kiyokazu
authors.initials: K
authors.lastname: Kokaji
authors.forename: Ryo
authors.initials: R
authors.lastname: Aeba
authors.forename: Ryohei
authors.initials: R
authors.lastname: Yozu
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Yamagishi
authors.forename: Toshio
authors.initials: T
authors.lastname: Kitamura
authors.forename: Keiichi
authors.initials: K
authors.lastname: Fukuda
authors.forename: Masaki
authors.initials: M
authors.lastname: Ieda
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D005786:Gene Expression Regulation; D006801:Humans; D007223:Infant; D008297:Male; D051379:Mice; D008875:Middle Aged; D009124:Muscle Proteins / Q000096:biosynthesis* / Q000235:genetics; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism*; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D009124:Muscle Proteins; D014157:Transcription Factors
keywords: cardiogenesis; cell fate conversion; regeneration