Detyrosinated microtubules provide mechanical resistance that can impede the motion of contracting cardiomyocytes. However, the functional effects of microtubule detyrosination in heart failure or in human hearts have not previously been studied. Here, we utilize mass spectrometry and single-myocyte mechanical assays to characterize changes to the cardiomyocyte cytoskeleton and their functional consequences in human heart failure. Proteomic analysis of left ventricle tissue reveals a consistent upregulation and stabilization of intermediate filaments and microtubules in failing human hearts. As revealed by super-resolution imaging, failing cardiomyocytes are characterized by a dense, heavily detyrosinated microtubule network, which is associated with increased myocyte stiffness and impaired contractility. Pharmacological suppression of detyrosinated microtubules lowers the viscoelasticity of failing myocytes and restores 40–50% of lost contractile function; reduction of microtubule detyrosination using a genetic approach also softens cardiomyocytes and improves contractile kinetics. Together, these data demonstrate that a modified cytoskeletal network impedes contractile function in cardiomyocytes from failing human hearts and that targeting detyrosinated microtubules could represent a new inotropic strategy for improving cardiac function.
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We would like thank the Quantitative Proteomics Resource Core of the Perelman School of Medicine and the Penn Center for Musculoskeletal Disorders Histology Core at the University of Pennsylvania. This work was supported by funding from the National Institutes of Health (NIH) R01-HL133080 to B.L.P. and T32 R05346-09 to P.R., the American Heart Association 17POST33440043 to M.A.C., and by the Center for Engineering MechanoBiology through a grant from the National Science Foundation’s Science and Technology Center program: 15-48571. The procurement of human heart tissue was enabled by grants from the NIH (HL089847 and HL105993) to K.B.M.
Supplementary Figures 1–6
Supplementary Tables 1–6
Skeletonized 3D reconstruction of the microtubule network from Airyscan imaging of a failing human myocyte (DCM) labeled with Tyr-tubulin antibody to demonstrate the density of the microtubule network
Microtubule buckling in a contracting non-failing myocyte infected with AdV-EMTB for 48 h
Microtubule buckling in a contracting failing myocyte (DCM) infected with AdV-EMTB for 48 h
Spreadsheet of mass spectrometry data
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Nature Reviews Cardiology (2018)