Increased tissue stiffness triggers contractile dysfunction and telomere shortening in dystrophic cardiomyocytes

Alex C Y Chang; Gaspard Pardon; Andrew C H Chang; Haodi Wu; Sang-Ging Ong; Asuka Eguchi; Sara Ancel; Colin Holbrook; John Ramunas; Alexandre J S Ribeiro; Edward L LaGory; Honghui Wang; Kassie Koleckar; Amato Giaccia; David L Mack; Martin K Childers; Chris Denning; John W Day; Joseph C Wu; Beth L Pruitt; Helen M Blau

doi:10.1016/j.stemcr.2021.04.018

Increased tissue stiffness triggers contractile dysfunction and telomere shortening in dystrophic cardiomyocytes

Stem Cell Reports. 2021 Sep 14;16(9):2169-2181. doi: 10.1016/j.stemcr.2021.04.018. Epub 2021 May 20.

Authors

Alex C Y Chang¹, Gaspard Pardon², Andrew C H Chang³, Haodi Wu⁴, Sang-Ging Ong⁴, Asuka Eguchi⁵, Sara Ancel⁵, Colin Holbrook⁵, John Ramunas⁵, Alexandre J S Ribeiro⁶, Edward L LaGory⁷, Honghui Wang⁸, Kassie Koleckar⁵, Amato Giaccia⁷, David L Mack⁹, Martin K Childers⁹, Chris Denning¹⁰, John W Day¹¹, Joseph C Wu⁴, Beth L Pruitt¹², Helen M Blau¹³

Affiliations

¹ Department of Cardiology and Shanghai Institute of Precision Medicine, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, A419, Bldg #2, 115 Jinzun Road, Pudong New District, Shanghai 200125, China; Baxter Laboratory for Stem Cell Biology, Department of Microbiology and Immunology, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CCSR Room 4215, 269 Campus Drive, Stanford, CA 94305-5175, USA; Division of Cardiovascular Medicine, Stanford University School of Medicine, Stanford, CA, USA; Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA. Electronic address: alexchang@shsmu.edu.cn.
² Baxter Laboratory for Stem Cell Biology, Department of Microbiology and Immunology, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CCSR Room 4215, 269 Campus Drive, Stanford, CA 94305-5175, USA; Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA; Departments of Bioengineering and Mechanical Engineering, Stanford University, School of Engineering and School of Medicine, Stanford, CA, USA; Mechanical Engineering and Biomolecular Science and Engineering, University of California, Santa Barbara, CA, USA.
³ Baxter Laboratory for Stem Cell Biology, Department of Microbiology and Immunology, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CCSR Room 4215, 269 Campus Drive, Stanford, CA 94305-5175, USA; Division of Cardiovascular Medicine, Stanford University School of Medicine, Stanford, CA, USA.
⁴ Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA.
⁵ Baxter Laboratory for Stem Cell Biology, Department of Microbiology and Immunology, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CCSR Room 4215, 269 Campus Drive, Stanford, CA 94305-5175, USA.
⁶ Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA; Departments of Bioengineering and Mechanical Engineering, Stanford University, School of Engineering and School of Medicine, Stanford, CA, USA.
⁷ Division of Radiation and Cancer Biology, Department of Radiation Oncology, Stanford University, Stanford, CA, USA.
⁸ Department of Cardiology and Shanghai Institute of Precision Medicine, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, A419, Bldg #2, 115 Jinzun Road, Pudong New District, Shanghai 200125, China.
⁹ Department of Rehabilitation Medicine, Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA, USA.
¹⁰ Division of Cancer & Stem Cells, Biodiscovery Institute, University of Nottingham, University Park NG7 2RD, UK.
¹¹ Department of Neurology, Stanford University, Stanford, CA, USA.
¹² Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA; Departments of Bioengineering and Mechanical Engineering, Stanford University, School of Engineering and School of Medicine, Stanford, CA, USA; Mechanical Engineering and Biomolecular Science and Engineering, University of California, Santa Barbara, CA, USA.
¹³ Baxter Laboratory for Stem Cell Biology, Department of Microbiology and Immunology, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CCSR Room 4215, 269 Campus Drive, Stanford, CA 94305-5175, USA; Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA. Electronic address: hblau@stanford.edu.

Abstract

Duchenne muscular dystrophy (DMD) is a rare X-linked recessive disease that is associated with severe progressive muscle degeneration culminating in death due to cardiorespiratory failure. We previously observed an unexpected proliferation-independent telomere shortening in cardiomyocytes of a DMD mouse model. Here, we provide mechanistic insights using human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Using traction force microscopy, we show that DMD hiPSC-CMs exhibit deficits in force generation on fibrotic-like bioengineered hydrogels, aberrant calcium handling, and increased reactive oxygen species levels. Furthermore, we observed a progressive post-mitotic telomere shortening in DMD hiPSC-CMs coincident with downregulation of shelterin complex, telomere capping proteins, and activation of the p53 DNA damage response. This telomere shortening is blocked by blebbistatin, which inhibits contraction in DMD cardiomyocytes. Our studies underscore the role of fibrotic stiffening in the etiology of DMD cardiomyopathy. In addition, our data indicate that telomere shortening is progressive, contraction dependent, and mechanosensitive, and suggest points of therapeutic intervention.

Keywords: DMD; dilated cardiomyopathy; fibrosis; hiPSC-CM; telomere.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Biomarkers
Cardiomyopathies / etiology
Cardiomyopathies / pathology
Cardiomyopathies / physiopathology
Cell Differentiation
Cells, Cultured
Cellular Microenvironment / drug effects
Culture Media, Conditioned / metabolism
Culture Media, Conditioned / pharmacology
Fibrosis
Fluorescent Antibody Technique
Gene Expression
Humans
Immunophenotyping
Induced Pluripotent Stem Cells / cytology
Induced Pluripotent Stem Cells / metabolism
Mechanical Phenomena
Muscular Dystrophies / genetics*
Muscular Dystrophies / pathology
Muscular Dystrophies / physiopathology*
Muscular Dystrophy, Duchenne / etiology
Muscular Dystrophy, Duchenne / pathology
Muscular Dystrophy, Duchenne / physiopathology
Myocardial Contraction / drug effects
Myocardial Contraction / genetics*
Myocytes, Cardiac / metabolism*
Telomere Shortening / genetics*

Substances

Biomarkers
Culture Media, Conditioned

Abstract

Publication types

MeSH terms

Substances

Grants and funding