iPS programmed without c-MYC yield proficient cardiogenesis for functional heart chimerism.
BACKGROUND
Induced pluripotent stem cells (iPS) allow derivation of pluripotent progenitors from somatic sources. Originally, iPS were induced by a stemness-related gene set that included the c-MYC oncogene.


OBJECTIVE
Here, we determined from embryo to adult the cardiogenic proficiency of iPS programmed without c-MYC, a cardiogenicity-associated transcription factor.


RESULTS
Transgenic expression of 3 human stemness factors SOX2, OCT4, and KLF4 here reset murine fibroblasts to the pluripotent ground state. Transduction without c-MYC reversed cellular ultrastructure into a primitive archetype and induced stem cell markers generating 3-germ layers, all qualifiers of acquired pluripotency. Three-factor induced iPS (3F-iPS) clones reproducibly demonstrated cardiac differentiation properties characterized by vigorous beating activity of embryoid bodies and robust expression of cardiac Mef2c, alpha-actinin, connexin43, MLC2a, and troponin I. In vitro isolated iPS-derived cardiomyocytes demonstrated functional excitation-contraction coupling. Chimerism with 3F-iPS derived by morula-stage diploid aggregation was sustained during prenatal heart organogenesis and contributed in vivo to normal cardiac structure and overall performance in adult tumor-free offspring.


CONCLUSIONS
Thus, 3F-iPS bioengineered without c-MYC achieve highest stringency criteria for bona fide cardiogenesis enabling reprogrammed fibroblasts to yield de novo heart tissue compatible with native counterpart throughout embryological development and into adulthood.
Martinez-Fernandez
Almudena
A
Marriott Heart Disease Research Program, Division of Cardiovascular Diseases, Departments of Medicine, Molecular Pharmacology and Experimental Therapeutics, and Medical Genetics, Mayo Clinic, Rochester, Minn. 55905, USA.
Nelson
Timothy J
TJ
Yamada
Satsuki
S
Reyes
Santiago
S
Alekseev
Alexey E
AE
Perez-Terzic
Carmen
C
Ikeda
Yasuhiro
Y
Terzic
Andre
A
D000185:Actinin / Q000378:metabolism; D000200:Action Potentials; D000818:Animals; D020013:Calcium Signaling; D019070:Cell Lineage; D054338:Cell Transdifferentiation / Q000235:genetics; D002478:Cells, Cultured; D046528:Chimerism; D018031:Connexin 43 / Q000378:metabolism; D046149:Embryo Culture Techniques; D005260:Female; D005347:Fibroblasts / Q000378:metabolism* / Q000648:ultrastructure; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics / Q000378:metabolism*; D064326:MEF2 Transcription Factors; D051379:Mice; D008810:Mice, Inbred C57BL; D008819:Mice, Nude; D009200:Myocardial Contraction / Q000235:genetics; D032383:Myocytes, Cardiac / Q000378:metabolism* / Q000648:ultrastructure; D018006:Myogenic Regulatory Factors / Q000378:metabolism; D018994:Myosin Light Chains / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism*; D038081:Organogenesis; D039904:Pluripotent Stem Cells / Q000378:metabolism* / Q000648:ultrastructure; D011247:Pregnancy; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism*; D023822:Tissue Engineering / Q000379:methods; D014161:Transduction, Genetic; D019210:Troponin I / Q000378:metabolism
D018031:Connexin 43; C543076:GJA1 protein, mouse; C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D064326:MEF2 Transcription Factors; C578041:Mef2c protein, mouse; C524495:Mlc2a protein, mouse; D018006:Myogenic Regulatory Factors; D018994:Myosin Light Chains; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D016271:Proto-Oncogene Proteins c-myc; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; D019210:Troponin I; D000185:Actinin
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