Cardiac diseases are fueled by extracellular matrix (ECM) remodelling. Together with the altered ECM chemical composition, the mechanical turmoil associated with ECM maladaptive remodelling in the pathological heart drives the shuttling of Yes Associated Protein 1 (YAP1) into cardiomyocyte (CM) nuclei that results either in cell cycle re-entry or cardiomyocyte hypertrophy. The mechanism of YAP1 reactivation and factors driving qualitatively different cellular outcomes is not well understood. Here we employed mechanical actuation as a proxy reproducing ECM remodelling in vitro to trigger YAP1 nuclear shuttling in contractile cardiomyocytes derived from human embryonic and induced pluripotent stem cells (hPSCs). By using hPSC lines in which YAP1 expression has been genetically depleted, super-resolution microscopy and electrophysiological measurements, we show that ECM-triggered nuclear presence of endogenous YAP1 contributes to cardiomyocyte maturation, participates in the formation and alignment of myofibrils, as well as in the maturation of their electrophysiological properties and calcium dynamics. We eventually exploit engineered heart tissues (EHTs) to demonstrate that the net effect of YAP1 deficiency in cardiomyocytes is the inability to respond to physiological stimuli by compensatory growth that results in reduced force development. These results suggest that the re-activation of endogenous YAP1 following ECM maladaptive remodelling promotes cardiomyocyte contractility by restructuring the sarcomere apparatus and the maturation of electrophysiological properties via transcriptionally dependent and independent mechanisms.

Faculty of Medicine Masaryk University Brno Czech Republic

Helmholtz Institute for Biomedical Engineering RWTH Aachen University Medical Faculty Aachen Germany

Institute for Stem Cell Biology RWTH Aachen University Medical School Aachen Germany

Institute of Physiology University of Bern Bern Switzerland

Institute of Theoretical and Applied Mechanics of the Czech Academy of Sciences Prague Czech Republic

International Clinical Research Center St Anne's University Hospital Brno Czech Republic

National Heart and Lung Institute Imperial College London London UK

School of Cardiovascular and Metabolic Medicine and Sciences King's College London London UK

Università degli Studi di Milano Bicocca Dept of Biotechnology and Biosciences Milan Italy

Università degli Studi di Milano Milan Italy

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