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Biomechanical Impact of Pathogenic MYBPC3 Truncation Variant Revealed by Dynamically Tuning In Vitro Afterload

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Abstract

Engineered cardiac microtissues were fabricated using pluripotent stem cells with a hypertrophic cardiomyopathy associated c. 2827 C>T; p.R943x truncation variant in myosin binding protein C (MYBPC3+/−). Microtissues were mounted on iron-incorporated cantilevers, allowing manipulations of cantilever stiffness using magnets, enabling examination of how in vitro afterload affects contractility. MYPBC3+/− microtissues developed augmented force, work, and power when cultured with increased in vitro afterload when compared with isogenic controls in which the MYBPC3 mutation had been corrected (MYPBC3+/+(ed)), but weaker contractility when cultured with lower in vitro afterload. After initial tissue maturation, MYPBC3+/− CMTs exhibited increased force, work, and power in response to both acute and sustained increases of in vitro afterload. Together, these studies demonstrate that extrinsic biomechanical challenges potentiate genetically-driven intrinsic increases in contractility that may contribute to clinical disease progression in patients with HCM due to hypercontractile MYBPC3 variants.

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Data Availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Abbreviations

CMT:

Cardiac microtissues

HCM:

Hypertrophic cardiomyopathy

Hu-iPSC-FBs:

Human-induced pluripotent stem cell fibroblasts

Hu-iPSC-CMs:

Human-induced pluripotent stem cell cardiomyocytes

iPSC:

Induced pluripotent stem cells

MRE:

Magnetorheological elastomer

MYPBC3:

Myosin binding protein C

PDMS:

Polydimethylsiloxane

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Funding

This research was supported by a grant from the State of Pennsylvania Department of Health, with additional support from NIH/NCATS award number TL1TR001880 to A.R. and C.E.L.; the Sidney Pestka M’61 Term Fellowship to A.R.; support from NIH T32-HL007843 to B.W.L.; American Heart Association 19CDA34770040 to C.K.L.; NIH/NHLBI grant R01 HL126527 and R01 HL130020 to J.C.W.; the Center for Engineering Mechanobiology (CEMB) under grant agreement CMMI: 15-48571 and a Provost Postdoctoral Fellowship to A.I.B.; and NIH/NHLBI grant R01-HL149891, the Leducq Foundation TNE ID#673168 and the Gordon and Llura Gund Family Fund to K.B.M.

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Authors and Affiliations

  1. Perelman School of Medicine, University of Pennsylvania, Smilow Center for Translational Research, 3400 Civic Center Boulevard, 11-101, Philadelphia, PA, 19104, USA

    Abhinay Ramachandran, Carissa E. Livingston & Kenneth B. Margulies

  2. Cardiovascular Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA

    Alexia Vite, Benjamin W. Lee & Kenneth B. Margulies

  3. Department of Biomedical Engineering, University of Delaware, Newark, DE, 19716, USA

    Elise A. Corbin

  4. Department of Materials Science and Engineering, University of Delaware, Newark, DE, 19716, USA

    Elise A. Corbin

  5. Nemours/Alfred I. duPont Hospital for Children, Wilmington, DE, 19803, USA

    Elise A. Corbin

  6. Department of Mechanical Engineering and Applied Mechanics, School of Engineering and Applied Sciences, University of Pennsylvania, Philadelphia, PA, 19104, USA

    Alexander I. Bennett & Kevin T. Turner

  7. Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, 94305, USA

    Chi Keung Lam & Joseph C. Wu

  8. Department of Biological Sciences, University of Delaware, Newark, DE, 19716, USA

    Chi Keung Lam

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  1. Abhinay Ramachandran
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  10. Kenneth B. Margulies
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Corresponding author

Correspondence to Kenneth B. Margulies.

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Human Subjects/Informed Consent/Animal Subjects Statement

While no human studies were carried out by the authors for this article, we utilized two cell lines of Hu-iPSC-CMs from the Stanford Cardiovascular Institute that were employed in the previous studies [17]. As described in Seeger et al., the protocol for induced pluripotent stem cell generation and cardiomyocyte differentiation were in accordance with the ethical standards of and approved by the Stanford Institutional Review Board (IRB) and Stem Cell Research Oversight (SCRO) Committee, and with the Helsinki Declaration of 1975, as revised in 2000. iPSC lines were obtained from patients with informed consent [17]. All procedures are followed of the responsible committee on human experimentation (institutional and national). Informed consent was obtained from all patients included in the study. No animal studies were carried out.

Competing Interests

Dr. Kenneth Margulies receives consulting fees for serving on the advisory board for Bristol-Myers-Squibb (myosin inhibitors).

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Associate Editor Paul J. R. Barton oversaw the review of this article

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Ramachandran, A., Livingston, C.E., Vite, A. et al. Biomechanical Impact of Pathogenic MYBPC3 Truncation Variant Revealed by Dynamically Tuning In Vitro Afterload. J. of Cardiovasc. Trans. Res. 16, 828–841 (2023). https://doi.org/10.1007/s12265-022-10348-4

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