YAP1 Contributes to The Development of Contractile Force and Sarcomere Maturation in Human Pluripotent Stem Cell Derived Cardiomyocytes

Vinarsky, V.; Pagliari, S.; Martino, F.; Mazzotti, C.; Jirakova, K.; Garlikova, Z.; Di Iuri, E.; Kytyr, D.; Benzoni, P.; Arici, M.; Metallo, A.; Zeevaert, K.; Wagner, W.; Rocchetti, M.; Forte, G.

doi:10.1101/2024.07.02.601803

bioRxiv · 10.1101/2024.07.02.601803

YAP1 Contributes to The Development of Contractile Force and Sarcomere Maturation in Human Pluripotent Stem Cell Derived Cardiomyocytes

Vinarsky, V.·Pagliari, S.·Martino, F.·Mazzotti, C.·Jirakova, K.·Garlikova, Z.·Di Iuri, E.·Kytyr, D.·Benzoni, P.·Arici, M.·Metallo, A.·Zeevaert, K.·Wagner, W.·Rocchetti, M.·Forte, G.

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Abstract

BackgroundPerspective regenerative therapies for injured myocardium focus on reactivating developmental processes to regenerate damaged contractile tissue. In animal models, the Hippo pathway was shown to improve heart regeneration after myocardial infarction, possibly by expanding the pool of cardiomyocytes. We hypothesized that activating the Hippo pathways downstream effector, Yes Associated Protein (YAP1), may have effects beyond promoting proliferation in human cardiomyocytes. We have therefore investigated how YAP1 influences human cardiomyocyte maturation, sarcomere structure, electrophysiological properties, their response to mechanical stimuli, intracellular Ca2+'' dynamics and force development using models of cardiomyocytes derived from pluripotent stem cells. MethodsWe employed genetic models of YAP1 deficient human embryonic and induced pluripotent stem cells, cardiomyocyte differentiation, compliant cultivation substrates, mechanical actuation, ECM deposition, super resolution microscopy, electrophysiological measurements and engineered heart tissues (EHTs) to characterize the effects of YAP1 deficiency on cardiomyocytes during maturation. We also used full length YAP1 re- expression to rescue the effects of YAP1 deficiency in contracting cardiomyocytes. ResultsYAP1 contributes to cardiomyocyte maturation, participates in the formation and alignment of myofibrils, as well as in the maturation of electrophysiological properties. The net effect of YAP1 deficiency in cardiomyocytes is the inability to respond to physiological stimuli by compensatory growth resulting in reduced force development. Additionally, YAP1 reactivation in contracting cardiomyocytes leads to rescue of myofibril maturation. ConclusionsThis research demonstrates that YAP1 activity is essential to promote cardiomyocyte maturation, contractility, and response to regeneration inducing stimuli.

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Vinarsky, V., Pagliari, S., Martino, F., Mazzotti, C., Jirakova, K., Garlikova, Z., Di Iuri, E., Kytyr, D., Benzoni, P., Arici, M., Metallo, A., Zeevaert, K., Wagner, W., Rocchetti, M., Forte, G.. 2024-07-05. YAP1 Contributes to The Development of Contractile Force and Sarcomere Maturation in Human Pluripotent Stem Cell Derived Cardiomyocytes. https://doi.org/10.1101/2024.07.02.601803

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