RRC ID 29211
Author 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.
Title Induction of cardiomyocyte-like cells in infarct hearts by gene transfer of Gata4, Mef2c, and Tbx5.
Journal Circ. Res.
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.
Volume 111(9)
Pages 1147-56
Published 2012-10-12
DOI 10.1161/CIRCRESAHA.112.271148
PII CIRCRESAHA.112.271148
PMID 22931955
MeSH Animals Cell Differentiation / genetics* Cell Differentiation / physiology Fibroblasts / pathology GATA4 Transcription Factor / genetics* GATA4 Transcription Factor / physiology Gene Transfer Techniques* Green Fluorescent Proteins / genetics MEF2 Transcription Factors Male Mice Mice, Inbred ICR Mice, Nude Mice, Transgenic Models, Animal Myocardial Infarction / pathology* Myocardial Infarction / physiopathology Myocytes, Cardiac / pathology* Myocytes, Cardiac / physiology Myogenic Regulatory Factors / genetics* Myogenic Regulatory Factors / physiology Regeneration / genetics Regeneration / physiology Retroviridae / genetics T-Box Domain Proteins / genetics* T-Box Domain Proteins / physiology
IF 15.211
Times Cited 98
WOS Category PERIPHERAL VASCULAR DISEASE HEMATOLOGY CARDIAC & CARDIOVASCULAR SYSTEMS
Resource
DNA material pMXs-GMT2A (RDB12141) pMXs-TMG2A (RDB12142)