Sarcomere function activates a p53-dependent DNA damage response that promotes polyploidization and limits in vivo cell engraftment
Anthony M. Pettinato,
Dasom Yoo,
Jennifer VanOudenhove,
Yu-Sheng Chen,
Rachel Cohn,
Feria A. Ladha,
Xiulan Yang,
Ketan Thakar,
Robert Romano,
Nicolas Legere,
Emily Meredith,
Paul Robson,
Michael Regnier,
Justin L. Cotney,
Charles E. Murry,
J. Travis Hinson
Affiliations
Anthony M. Pettinato
Department of Genetics and Genome Sciences, UConn Health, Farmington, CT 06030, USA
Dasom Yoo
Department of Bioengineering, University of Washington, Seattle, WA 98109, USA
Jennifer VanOudenhove
Department of Genetics and Genome Sciences, UConn Health, Farmington, CT 06030, USA
Yu-Sheng Chen
The Jackson Laboratory for Genomic Medicine, Farmington, CT 06032, USA
Rachel Cohn
The Jackson Laboratory for Genomic Medicine, Farmington, CT 06032, USA
Feria A. Ladha
Department of Genetics and Genome Sciences, UConn Health, Farmington, CT 06030, USA
Xiulan Yang
Center for Cardiovascular Biology and Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA 98109, USA
Ketan Thakar
The Jackson Laboratory for Genomic Medicine, Farmington, CT 06032, USA
Robert Romano
The Jackson Laboratory for Genomic Medicine, Farmington, CT 06032, USA
Nicolas Legere
The Jackson Laboratory for Genomic Medicine, Farmington, CT 06032, USA
Emily Meredith
The Jackson Laboratory for Genomic Medicine, Farmington, CT 06032, USA
Paul Robson
The Jackson Laboratory for Genomic Medicine, Farmington, CT 06032, USA
Michael Regnier
Department of Bioengineering, University of Washington, Seattle, WA 98109, USA
Justin L. Cotney
Department of Genetics and Genome Sciences, UConn Health, Farmington, CT 06030, USA
Charles E. Murry
Department of Bioengineering, University of Washington, Seattle, WA 98109, USA; Center for Cardiovascular Biology and Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA 98109, USA; Department of Pathology, University of Washington, Seattle, WA 98109, USA; Department of Medicine/Cardiology, University of Washington, Seattle, WA 98109, USA
J. Travis Hinson
Department of Genetics and Genome Sciences, UConn Health, Farmington, CT 06030, USA; The Jackson Laboratory for Genomic Medicine, Farmington, CT 06032, USA; Corresponding author
Summary: Human cardiac regeneration is limited by low cardiomyocyte replicative rates and progressive polyploidization by unclear mechanisms. To study this process, we engineer a human cardiomyocyte model to track replication and polyploidization using fluorescently tagged cyclin B1 and cardiac troponin T. Using time-lapse imaging, in vitro cardiomyocyte replication patterns recapitulate the progressive mononuclear polyploidization and replicative arrest observed in vivo. Single-cell transcriptomics and chromatin state analyses reveal that polyploidization is preceded by sarcomere assembly, enhanced oxidative metabolism, a DNA damage response, and p53 activation. CRISPR knockout screening reveals p53 as a driver of cell-cycle arrest and polyploidization. Inhibiting sarcomere function, or scavenging ROS, inhibits cell-cycle arrest and polyploidization. Finally, we show that cardiomyocyte engraftment in infarcted rat hearts is enhanced 4-fold by the increased proliferation of troponin-knockout cardiomyocytes. Thus, the sarcomere inhibits cell division through a DNA damage response that can be targeted to improve cardiomyocyte replacement strategies.
