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Extracellular stiffness induces contractile dysfunction in adult cardiomyocytes via cell-autonomous and microtubule-dependent mechanisms

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Abstract

The mechanical environment of the myocardium has a potent effect on cardiomyocyte form and function, yet an understanding of the cardiomyocyte responses to extracellular stiffening remains incomplete. We therefore employed a cell culture substrate with tunable stiffness to define the cardiomyocyte responses to clinically relevant stiffness increments in the absence of cell–cell interactions. When cultured on substrates magnetically actuated to mimic the stiffness of diseased myocardium, isolated rat adult cardiomyocytes exhibited a time-dependent reduction of sarcomere shortening, characterized by slowed contraction and relaxation velocity, and alterations of the calcium transient. Cardiomyocytes cultured on stiff substrates developed increases in viscoelasticity and microtubule detyrosination in association with early increases in the α-tubulin detyrosinating enzyme vasohibin-2 (Vash2). We found that knockdown of Vash2 was sufficient to preserve contractile performance as well as calcium transient properties in the presence of extracellular substrate stiffening. Orthogonal prevention of detyrosination by overexpression of tubulin tyrosine ligase (TTL) was also able to preserve contractility and calcium homeostasis. These data demonstrate that a pathologic increment of extracellular stiffness induces early, cell-autonomous remodeling of adult cardiomyocytes that is dependent on detyrosination of α-tubulin.

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Funding

This research was supported by funding from NIH/NHLBI R01-HL149891-01 to K.B.M and B.L.P., R01-HL133080 to B.L.P, a Leducq Fondation award TNE ID#: 673168 to B.L.P and K.B.M., Gund Family Fund support to K.B.M., and AHA CDA 856504 to M.A.C.

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Author notes

  1. Alexia Vite and Matthew A. Caporizzo contributed equally to this work.

Authors and Affiliations

  1. Department of Medicine and Cardiovascular Institute, Perelman School of Medicine, University of Pennsylvania, 3400 Civic Center Boulevard, Smillow TRC 11-101, Philadelphia, PA, 19104, USA

    Alexia Vite, Jeffrey Brandimarto, Quentin McAfee, Carissa E. Livingston & Kenneth B. Margulies

  2. Department of Molecular Physiology and Biophysics, University of Vermont’s Larner College of Medicine, Burlington, VT, USA

    Matthew A. Caporizzo

  3. Departments of Biomedical Engineering and Materials Science, University of Delaware, Newark, DE, USA

    Elise A. Corbin

  4. Department of Physiology and Pennsylvania Muscle Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA

    Benjamin L. Prosser & Kenneth B. Margulies

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  1. Alexia Vite
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Contributions

AV, MAC, BLP, and KBM developed the research strategy. AV, MAC, EAC BLP, and KBM participated in the design of experiments. AV, MAC, EAC, JB, QM and CEL performed the experiments. All authors participated in the writing and review of the manuscript.

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Correspondence to Kenneth B. Margulies.

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Conflict of interest

Drs. Prosser and Margulies report significant financial interests: invention disclosure/patent; inventor: US patent application No.15/959,181 USA 2018, composition and methods for improving heart function and treating heart failure.

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Vite, A., Caporizzo, M.A., Corbin, E.A. et al. Extracellular stiffness induces contractile dysfunction in adult cardiomyocytes via cell-autonomous and microtubule-dependent mechanisms. Basic Res Cardiol 117, 41 (2022). https://doi.org/10.1007/s00395-022-00952-5

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  • DOI: https://doi.org/10.1007/s00395-022-00952-5

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