Contractile Defect Caused by Mutation in MYBPC3 Revealed under Conditions Optimized for Human PSC-Cardiomyocyte Function.
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.
Department of Anatomy and Embryology, Leiden University Medical Center, 2300 RC Leiden, the Netherlands.
Matthew J
MJ
Birket
Department of Anatomy and Embryology, Leiden University Medical Center, 2300 RC Leiden, the Netherlands.
Marcelo C
MC
Ribeiro
Department of Anatomy and Embryology, Leiden University Medical Center, 2300 RC Leiden, the Netherlands.
Georgios
G
Kosmidis
Department of Anatomy and Embryology, Leiden University Medical Center, 2300 RC Leiden, the Netherlands.
Dorien
D
Ward
Department of Anatomy and Embryology, Leiden University Medical Center, 2300 RC Leiden, the Netherlands.
Ana Rita
AR
Leitoguinho
Department of Anatomy and Embryology, Leiden University Medical Center, 2300 RC Leiden, the Netherlands.
Vera
V
van de Pol
Department of Anatomy and Embryology, Leiden University Medical Center, 2300 RC Leiden, the Netherlands; Department of Cardiology, Leiden University Medical Center, 2300 RC Leiden, the Netherlands.
Cheryl
C
Dambrot
Department of Anatomy and Embryology, Leiden University Medical Center, 2300 RC Leiden, the Netherlands.
Harsha D
HD
Devalla
Department of Anatomy and Embryology, Leiden University Medical Center, 2300 RC Leiden, the Netherlands.
Richard P
RP
Davis
Erasmus Medical Center, 3015 GE Rotterdam, the Netherlands.
Pier G
PG
Mastroberardino
Department of Cardiology, Leiden University Medical Center, 2300 RC Leiden, the Netherlands.
Douwe E
DE
Atsma
Department of Anatomy and Embryology, Leiden University Medical Center, 2300 RC Leiden, the Netherlands.
Robert
R
Passier
Department of Anatomy and Embryology, Leiden University Medical Center, 2300 RC Leiden, the Netherlands. Electronic address: c.l.mummery@lumc.nl.
Christine L
CL
Mummery
D000818:Animals; D002312:Cardiomyopathy, Hypertrophic; D002352:Carrier Proteins / Q000235:genetics*; D002454:Cell Differentiation; D002460:Cell Line; D004594:Electrophysiology; D005434:Flow Cytometry; D006801:Humans; D051379:Mice; D009154:Mutation / Q000235:genetics; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism*; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D000099163:Myosin Binding Protein C
D002352:Carrier Proteins; D000099163:Myosin Binding Protein C