title: Fibroblast Growth Factors and Vascular Endothelial Growth Factor Promote Cardiac Reprogramming under Defined Conditions.
abstract: Fibroblasts can be directly reprogrammed into cardiomyocyte-like cells (iCMs) by overexpression of cardiac transcription factors, including Gata4, Mef2c, and Tbx5; however, this process is inefficient under serum-based culture conditions, in which conversion of partially reprogrammed cells into fully reprogrammed functional iCMs has been a major hurdle. Here, we report that a combination of fibroblast growth factor (FGF) 2, FGF10, and vascular endothelial growth factor (VEGF), termed FFV, promoted cardiac reprogramming under defined serum-free conditions, increasing spontaneously beating iCMs by 100-fold compared with those under conventional serum-based conditions. Mechanistically, FFV activated multiple cardiac transcriptional regulators and converted partially reprogrammed cells into functional iCMs through the p38 mitogen-activated protein kinase and phosphoinositol 3-kinase/AKT pathways. Moreover, FFV enabled cardiac reprogramming with only Mef2c and Tbx5 through the induction of cardiac reprogramming factors, including Gata4. Thus, defined culture conditions promoted the quality of cardiac reprogramming, and this finding provides new insight into the mechanism of cardiac reprogramming.
authors.affiliation: Department of Cardiology, Keio University School of Medicine, Tokyo 160-8582, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Yamakawa
authors.affiliation: Department of Cardiology, Keio University School of Medicine, Tokyo 160-8582, Japan.
authors.forename: Naoto
authors.initials: N
authors.lastname: Muraoka
authors.affiliation: Department of Cardiology, Keio University School of Medicine, Tokyo 160-8582, Japan.
authors.forename: Kazutaka
authors.initials: K
authors.lastname: Miyamoto
authors.affiliation: Department of Cardiology, Keio University School of Medicine, Tokyo 160-8582, Japan.
authors.forename: Taketaro
authors.initials: T
authors.lastname: Sadahiro
authors.affiliation: Department of Cardiology, Keio University School of Medicine, Tokyo 160-8582, Japan.
authors.forename: Mari
authors.initials: M
authors.lastname: Isomi
authors.affiliation: Department of Cardiology, Keio University School of Medicine, Tokyo 160-8582, Japan.
authors.forename: Sho
authors.initials: S
authors.lastname: Haginiwa
authors.affiliation: Department of Cardiology, Keio University School of Medicine, Tokyo 160-8582, Japan.
authors.forename: Hidenori
authors.initials: H
authors.lastname: Kojima
authors.affiliation: Department of Cardiology, Keio University School of Medicine, Tokyo 160-8582, Japan.
authors.forename: Tomohiko
authors.initials: T
authors.lastname: Umei
authors.affiliation: Department of Cardiology, Keio University School of Medicine, Tokyo 160-8582, Japan.
authors.forename: Mizuha
authors.initials: M
authors.lastname: Akiyama
authors.affiliation: Department of Cardiology, Keio University School of Medicine, Tokyo 160-8582, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Kuishi
authors.affiliation: Department of Bio-Informational Pharmacology, Medical Research Institute, Tokyo Medical Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, Japan.
authors.forename: Junko
authors.initials: J
authors.lastname: Kurokawa
authors.affiliation: Department of Bio-Informational Pharmacology, Medical Research Institute, Tokyo Medical Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, Japan.
authors.forename: Tetsushi
authors.initials: T
authors.lastname: Furukawa
authors.affiliation: Department of Cardiology, Keio University School of Medicine, Tokyo 160-8582, Japan.
authors.forename: Keiichi
authors.initials: K
authors.lastname: Fukuda
authors.affiliation: Department of Cardiology, Keio University School of Medicine, Tokyo 160-8582, Japan; JST, CREST, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan. Electronic address: mieda@z8.keio.jp.
authors.forename: Masaki
authors.initials: M
authors.lastname: Ieda
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D000067470:Cellular Reprogramming Techniques / Q000379:methods*; D005346:Fibroblast Growth Factors / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D064326:MEF2 Transcription Factors / Q000235:genetics / Q000378:metabolism; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D015398:Signal Transduction; D020825:T-Box Domain Proteins / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: D064326:MEF2 Transcription Factors; C578041:Mef2c protein, mouse; D020825:T-Box Domain Proteins; C103172:T-box transcription factor 5; D042461:Vascular Endothelial Growth Factor A; D005346:Fibroblast Growth Factors; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D048051:p38 Mitogen-Activated Protein Kinases