The integration of in vitro cardiac tissue models, human induced pluripotent stem cells (hiPSCs) and genome-editing tools allows for the enhanced interrogation of physiological phenotypes and recapitulation of disease pathologies. Here, using a cardiac tissue model consisting of filamentous three-dimensional matrices populated with cardiomyocytes derived from healthy wild-type (WT) hiPSCs (WT hiPSC-CMs) or isogenic hiPSCs deficient in the sarcomere protein cardiac myosin-binding protein C (MYBPC3–/– hiPSC-CMs), we show that the WT microtissues adapted to the mechanical environment with increased contraction force commensurate to matrix stiffness, whereas the MYBPC3–/– microtissues exhibited impaired force development kinetics regardless of matrix stiffness and deficient contraction force only when grown on matrices with high fibre stiffness. Under mechanical overload, the MYBPC3–/– microtissues had a higher degree of calcium transient abnormalities, and exhibited an accelerated decay of calcium dynamics as well as calcium desensitization, which accelerated when contracting against stiffer fibres. Our findings suggest that MYBPC3 deficiency and the presence of environmental stresses synergistically lead to contractile deficits in cardiac tissues.
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This work was supported in part by the NIH NHLBI (R01HL096525, R01HL108677, U01HL100406 and U01HL098179), NIH NIBIB (R21EB021003) and NIH NCATS (UH3TR000487). Z.M. acknowledges support from NSF (1804875), American Heart Association postdoctoral fellowship (16POST27750031) and the Nappi Family Foundation Research Scholar Project. N.H. acknowledges support from NIH T32 (HL007544). M.A.M. acknowledges support from the Canadian Institute of Health Research Postdoctoral Fellowship (129844). B.S. acknowledges support from the California Institute for Regenerative Medicine (TBI-01197). We acknowledge assistance from the Berkeley CIRM/QB3 Shared Stem Cell Facility for flow cytometry, Biological Imaging Facility for confocal microscopy (supported by NIH S10 programme 1S10RR026866-01) and Biomolecular Nanotechnology Center for scanning electron microscopy. We thank E. Nora and P. Devine (Gladstone Institute of Cardiovascular Disease) for helpful discussions and advice regarding p300 expression analysis. The contents of this publication are solely the responsibility of the authors and do not necessarily represent the official views of the California Institute for Regenerative Medicine and/or other agencies of the State of California.
B.R.C. is a founder of Tenaya Therapeutics, a company focused on finding treatments for heart failure, including the use of CRISPR interference to interrogate genetic cardiomyopathies. B.R.C. holds equity in Tenaya Therapeutics, and Tenaya Therapeutics provides research support for heart failure-related research to B.R.C. K.E.H. and N.H. have a financial relationship with Organos, and both themselves and the company may benefit from commercialization of the results of this research. The other authors declare no competing interests.
Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary methods, figures, tables, references and video captions
The fabrication process of filamentous matrices. One fibre was fabricated with one single shot of laser irradiation
Five days after seeding WT hiPSC-CMs into a 5 μm filamentous matrix, the cardiac microtissue formed and bent the fibres during the contraction
Time-dependent transient stress presented to the fibres was simulated using COMSOL during the cardiac microtissue contraction
Twenty days after seeding MYBPC3–/– hiPSC-CMs into a 10 μm filamentous matrix, the cardiac microtissue formed and bent the fibres during the contraction
Twenty days after seeding GCaMP6f-WT hiPS-CMs into a 10 μm filamentous matrix, the calcium dynamics was recorded under a fluorescent microscope
Twenty days post-differentiation, WT hiPSC-CMs beat as a sheet and were analysed by motion-tracking software to show the contraction vectors
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Ma, Z., Huebsch, N., Koo, S. et al. Contractile deficits in engineered cardiac microtissues as a result of MYBPC3 deficiency and mechanical overload. Nat Biomed Eng 2, 955–967 (2018). https://doi.org/10.1038/s41551-018-0280-4
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