Induction of cardiomyocyte-like cells in infarct hearts by gene transfer of Gata4, Mef2c, and Tbx5

Kohei Inagawa, Kazutaka Miyamoto, Hiroyuki Yamakawa, Naoto Muraoka, Taketaro Sadahiro, Tomohiko Umei, Rie Wada, Yoshinori Katsumata, Ruri Kaneda, Koji Nakade, Chitose Kurihara, Yuichi Obata, Koichi Miyake, Keiichi Fukuda, Masaki Ieda

Research output: Contribution to journalArticle

150 Citations (Scopus)

Abstract

RATIONALE: After myocardial infarction (MI), massive cell death in the myocardium initiates fibrosis and scar formation, leading to heart failure. We recently found that a combination of 3 cardiac transcription factors, Gata4, Mef2c, and Tbx5 (GMT), reprograms fibroblasts directly into functional cardiomyocytes in vitro. OBJECTIVE: To investigate whether viral gene transfer of GMT into infarcted hearts induces cardiomyocyte generation. METHODS AND RESULTS: Coronary artery ligation was used to generate MI in the mouse. In vitro transduction of GMT retrovirus converted cardiac fibroblasts from the infarct region into cardiomyocyte-like cells with cardiac-specific gene expression and sarcomeric structures. Injection of the green fluorescent protein (GFP) retrovirus into mouse hearts, immediately after MI, infected only proliferating noncardiomyocytes, mainly fibroblasts, in the infarct region. The GFP expression diminished after 2 weeks in immunocompetent mice but remained stable for 3 months in immunosuppressed mice, in which cardiac induction did not occur. In contrast, injection of GMT retrovirus into α-myosin heavy chain (αMHC)-GFP transgenic mouse hearts induced the expression of αMHC-GFP, a marker of cardiomyocytes, in 3% of virus-infected cells after 1 week. A pooled GMT injection into the immunosuppressed mouse hearts induced cardiac marker expression in retrovirus-infected cells within 2 weeks, although few cells showed striated muscle structures. To transduce GMT efficiently in vivo, we generated a polycistronic retrovirus expressing GMT separated by 2A "self-cleaving" peptides (3F2A). The 3F2A-induced cardiomyocyte-like cells in fibrotic tissue expressed sarcomeric α-actinin and cardiac troponin T and had clear cross striations. Quantitative RT-PCR also demonstrated that FACS-sorted 3F2A-transduced cells expressed cardiac-specific genes. CONCLUSIONS: GMT gene transfer induced cardiomyocyte-like cells in infarcted hearts.

Original languageEnglish
Pages (from-to)1147-1156
Number of pages10
JournalCirculation Research
Volume111
Issue number9
DOIs
Publication statusPublished - 2012 Oct 12

Fingerprint

Cardiac Myocytes
Retroviridae
Green Fluorescent Proteins
Genes
Myosin Heavy Chains
Fibroblasts
Myocardial Infarction
Injections
Transcription Factor 3
Actinin
Troponin T
Striated Muscle
Viral Genes
Transgenic Mice
Cicatrix
Ligation
Coronary Vessels
Myocardium
Fibrosis
Cell Death

Keywords

  • cardiomyocytes
  • fibroblasts
  • myocardial infarction
  • reprogramming
  • transcription factors

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Induction of cardiomyocyte-like cells in infarct hearts by gene transfer of Gata4, Mef2c, and Tbx5. / Inagawa, Kohei; Miyamoto, Kazutaka; Yamakawa, Hiroyuki; Muraoka, Naoto; Sadahiro, Taketaro; Umei, Tomohiko; Wada, Rie; Katsumata, Yoshinori; Kaneda, Ruri; Nakade, Koji; Kurihara, Chitose; Obata, Yuichi; Miyake, Koichi; Fukuda, Keiichi; Ieda, Masaki.

In: Circulation Research, Vol. 111, No. 9, 12.10.2012, p. 1147-1156.

Research output: Contribution to journalArticle

Inagawa, K, Miyamoto, K, Yamakawa, H, Muraoka, N, Sadahiro, T, Umei, T, Wada, R, Katsumata, Y, Kaneda, R, Nakade, K, Kurihara, C, Obata, Y, Miyake, K, Fukuda, K & Ieda, M 2012, 'Induction of cardiomyocyte-like cells in infarct hearts by gene transfer of Gata4, Mef2c, and Tbx5', Circulation Research, vol. 111, no. 9, pp. 1147-1156. https://doi.org/10.1161/CIRCRESAHA.112.271148
Inagawa, Kohei ; Miyamoto, Kazutaka ; Yamakawa, Hiroyuki ; Muraoka, Naoto ; Sadahiro, Taketaro ; Umei, Tomohiko ; Wada, Rie ; Katsumata, Yoshinori ; Kaneda, Ruri ; Nakade, Koji ; Kurihara, Chitose ; Obata, Yuichi ; Miyake, Koichi ; Fukuda, Keiichi ; Ieda, Masaki. / Induction of cardiomyocyte-like cells in infarct hearts by gene transfer of Gata4, Mef2c, and Tbx5. In: Circulation Research. 2012 ; Vol. 111, No. 9. pp. 1147-1156.
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AU - Yamakawa, Hiroyuki

AU - Muraoka, Naoto

AU - Sadahiro, Taketaro

AU - Umei, Tomohiko

AU - Wada, Rie

AU - Katsumata, Yoshinori

AU - Kaneda, Ruri

AU - Nakade, Koji

AU - Kurihara, Chitose

AU - Obata, Yuichi

AU - Miyake, Koichi

AU - Fukuda, Keiichi

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N2 - RATIONALE: After myocardial infarction (MI), massive cell death in the myocardium initiates fibrosis and scar formation, leading to heart failure. We recently found that a combination of 3 cardiac transcription factors, Gata4, Mef2c, and Tbx5 (GMT), reprograms fibroblasts directly into functional cardiomyocytes in vitro. OBJECTIVE: To investigate whether viral gene transfer of GMT into infarcted hearts induces cardiomyocyte generation. METHODS AND RESULTS: Coronary artery ligation was used to generate MI in the mouse. In vitro transduction of GMT retrovirus converted cardiac fibroblasts from the infarct region into cardiomyocyte-like cells with cardiac-specific gene expression and sarcomeric structures. Injection of the green fluorescent protein (GFP) retrovirus into mouse hearts, immediately after MI, infected only proliferating noncardiomyocytes, mainly fibroblasts, in the infarct region. The GFP expression diminished after 2 weeks in immunocompetent mice but remained stable for 3 months in immunosuppressed mice, in which cardiac induction did not occur. In contrast, injection of GMT retrovirus into α-myosin heavy chain (αMHC)-GFP transgenic mouse hearts induced the expression of αMHC-GFP, a marker of cardiomyocytes, in 3% of virus-infected cells after 1 week. A pooled GMT injection into the immunosuppressed mouse hearts induced cardiac marker expression in retrovirus-infected cells within 2 weeks, although few cells showed striated muscle structures. To transduce GMT efficiently in vivo, we generated a polycistronic retrovirus expressing GMT separated by 2A "self-cleaving" peptides (3F2A). The 3F2A-induced cardiomyocyte-like cells in fibrotic tissue expressed sarcomeric α-actinin and cardiac troponin T and had clear cross striations. Quantitative RT-PCR also demonstrated that FACS-sorted 3F2A-transduced cells expressed cardiac-specific genes. CONCLUSIONS: GMT gene transfer induced cardiomyocyte-like cells in infarcted hearts.

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