TY - JOUR
T1 - Contractile Defect Caused by Mutation in MYBPC3 Revealed under Conditions Optimized for Human PSC-Cardiomyocyte Function
AU - Birket, Matthew J.
AU - Ribeiro, Marcelo C.
AU - Kosmidis, Georgios
AU - Ward, Dorien
AU - Leitoguinho, Ana Rita
AU - van de Pol, Vera
AU - Dambrot, Cheryl
AU - Devalla, Harsha D.
AU - Davis, Richard P.
AU - Mastroberardino, Pier G.
AU - Atsma, Douwe E.
AU - Passier, Robert
AU - Mummery, Christine L.
N1 - Funding Information: Work is supported by Cardiovascular Research Netherlands (CVON HUSTCARE), Netherlands Institute of Regenerative Medicine (NIRM), the European Research Council (ERCAdG 323182 STEMCARDIOVASC), the Rembrandt Institute of Cardiovascular Science RICS, and the Netherlands Organization for Scientific Research (NWO-FOM FOM 09MMC02). Other support includes ZonMw-MKMD-40-42600-98-036 (R.P.), Netherlands Genomics Initiative NGI/NWO 05040202 (P.M.), and Marie Curie IRG 247918 (P.M.). The Seahorse Extracellular Flux Analyzer was purchased through the generous contribution of the Dorpmans-Wigmans Stichting. We thank Dr. Christian Freund and Simona van de Pas of the LUMC hiPSC Core Facility for the “Con1” hiPSC line, Dr. Sakthivel Sadayappan (Loyola University Chicago) for the cMyBP-C antibody, and Ajit Divakaruni (University of California, San Diego) for helpful advice with the design of Seahorse experiments. Publisher Copyright: © 2015 The Authors.
PY - 2015/10/27
Y1 - 2015/10/27
N2 - Maximizing baseline function of human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) is essential for their effective application in models of cardiac toxicity and disease. Here, we aimed to identify factors that would promote an adequate level of function to permit robust single-cell contractility measurements in a human induced pluripotent stem cell (hiPSC) model of hypertrophic cardiomyopathy (HCM). A simple screen revealed the collaborative effects of thyroid hormone, IGF-1 and the glucocorticoid analog dexamethasone on the electrophysiology, bioenergetics, and contractile force generation of hPSC-CMs. In this optimized condition, hiPSC-CMs with mutations in MYBPC3, a gene encoding myosin-binding protein C, which, when mutated, causes HCM, showed significantly lower contractile force generation than controls. This was recapitulated by direct knockdown of MYBPC3 in control hPSC-CMs, supporting a mechanism of haploinsufficiency. Modeling this disease in vitro using human cells is an important step toward identifying therapeutic interventions for HCM.
AB - Maximizing baseline function of human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) is essential for their effective application in models of cardiac toxicity and disease. Here, we aimed to identify factors that would promote an adequate level of function to permit robust single-cell contractility measurements in a human induced pluripotent stem cell (hiPSC) model of hypertrophic cardiomyopathy (HCM). A simple screen revealed the collaborative effects of thyroid hormone, IGF-1 and the glucocorticoid analog dexamethasone on the electrophysiology, bioenergetics, and contractile force generation of hPSC-CMs. In this optimized condition, hiPSC-CMs with mutations in MYBPC3, a gene encoding myosin-binding protein C, which, when mutated, causes HCM, showed significantly lower contractile force generation than controls. This was recapitulated by direct knockdown of MYBPC3 in control hPSC-CMs, supporting a mechanism of haploinsufficiency. Modeling this disease in vitro using human cells is an important step toward identifying therapeutic interventions for HCM.
UR - http://www.scopus.com/inward/record.url?scp=84945580024&partnerID=8YFLogxK
U2 - https://doi.org/10.1016/j.celrep.2015.09.025
DO - https://doi.org/10.1016/j.celrep.2015.09.025
M3 - Article
C2 - 26489474
SN - 2211-1247
VL - 13
SP - 733
EP - 745
JO - Cell reports
JF - Cell reports
IS - 4
ER -