Maturation of iPSC-derived cardiomyocytes in a heart-on-a-chip device enables modeling of dilated cardiomyopathy caused by R222Q-SCN5A mutation

Marianne Wauchop; Naimeh Rafatian; Yimu Zhao; Wenliang Chen; Mark Gagliardi; Stéphane Massé; Brian J Cox; Patrick Lai; Timothy Liang; Shira Landau; Stephanie Protze; Xiao Dong Gao; Erika Yan Wang; Kelvin Chan Tung; Zachary Laksman; Rick Xing Ze Lu; Gordon Keller; Kumaraswamy Nanthakumar; Milica Radisic; Peter H Backx

doi:10.1016/j.biomaterials.2023.122255

Maturation of iPSC-derived cardiomyocytes in a heart-on-a-chip device enables modeling of dilated cardiomyopathy caused by R222Q-SCN5A mutation

Biomaterials. 2023 Oct:301:122255. doi: 10.1016/j.biomaterials.2023.122255. Epub 2023 Jul 26.

Authors

Marianne Wauchop¹, Naimeh Rafatian², Yimu Zhao³, Wenliang Chen⁴, Mark Gagliardi⁵, Stéphane Massé⁶, Brian J Cox⁷, Patrick Lai⁶, Timothy Liang⁶, Shira Landau⁸, Stephanie Protze⁹, Xiao Dong Gao¹⁰, Erika Yan Wang⁸, Kelvin Chan Tung⁵, Zachary Laksman¹¹, Rick Xing Ze Lu⁸, Gordon Keller¹², Kumaraswamy Nanthakumar¹³, Milica Radisic¹⁴, Peter H Backx¹⁵

Affiliations

¹ Department of Physiology, Faculty of Medicine, University of Toronto, Toronto, ON, M5S 1A8, Canada.
² Division of Cardiology and Peter Munk Cardiac Center, University Health Network, Toronto, ON, M5G 1L7, Canada.
³ Toronto General Hospital Research Institute, Toronto, ON, M5G 2C4, Canada; Institute of Biomedical Engineering, University of Toronto, Toronto, ON, M5S 3G9, Canada.
⁴ Division of Cardiology and Peter Munk Cardiac Center, University Health Network, Toronto, ON, M5G 1L7, Canada; Department of Biology, York University, Toronto, ON, M3J 1P3, Canada.
⁵ McEwen Stem Cell Institute, University Health Network, Toronto, ON, M5G 1L7, Canada.
⁶ Division of Cardiology and Peter Munk Cardiac Center, University Health Network, Toronto, ON, M5G 1L7, Canada; Toronto General Hospital Research Institute, Toronto, ON, M5G 2C4, Canada; The Hull Family Cardiac Fibrillation Management Laboratory, Toronto General Hospital, Toronto, ON, M5G 2C4, Canada.
⁷ Department of Physiology, Faculty of Medicine, University of Toronto, Toronto, ON, M5S 1A8, Canada; Department of Obstetrics and Gynaecology, Faculty of Medicine, University of Toronto, Toronto, ON, M5S 1A8, Canada.
⁸ Institute of Biomedical Engineering, University of Toronto, Toronto, ON, M5S 3G9, Canada.
⁹ McEwen Stem Cell Institute, University Health Network, Toronto, ON, M5G 1L7, Canada; Department of Molecular Genetics, University of Toronto, Toronto, ON, M5S 1A8, Canada.
¹⁰ Department of Biology, York University, Toronto, ON, M3J 1P3, Canada.
¹¹ Department of Medicine, University of British Columbia, Vancouver, BC, V6E 1M7, Canada.
¹² McEwen Stem Cell Institute, University Health Network, Toronto, ON, M5G 1L7, Canada; Department of Medical Biophysics, University of Toronto, Toronto, ON, M5G 1L7, Canada.
¹³ Division of Cardiology and Peter Munk Cardiac Center, University Health Network, Toronto, ON, M5G 1L7, Canada; Toronto General Hospital Research Institute, Toronto, ON, M5G 2C4, Canada; The Hull Family Cardiac Fibrillation Management Laboratory, Toronto General Hospital, Toronto, ON, M5G 2C4, Canada. Electronic address: kumar.nanthakumar@uhn.ca.
¹⁴ Toronto General Hospital Research Institute, Toronto, ON, M5G 2C4, Canada; Institute of Biomedical Engineering, University of Toronto, Toronto, ON, M5S 3G9, Canada; Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, ON, Canada, M5S 3E5. Electronic address: m.radisic@utoronto.ca.
¹⁵ Division of Cardiology and Peter Munk Cardiac Center, University Health Network, Toronto, ON, M5G 1L7, Canada; Department of Biology, York University, Toronto, ON, M3J 1P3, Canada; Terrence Donnelly Centre for Cellular & Biomolecular Research, University of Toronto, Toronto, ON, M5S 3E1, Canada. Electronic address: p.backx@utoronto.ca.

PMID: 37651922
PMCID: PMC10942743 (available on 2024-10-01)
DOI: 10.1016/j.biomaterials.2023.122255

Abstract

To better understand sodium channel (SCN5A)-related cardiomyopathies, we generated ventricular cardiomyocytes from induced pluripotent stem cells obtained from a dilated cardiomyopathy patient harbouring the R222Q mutation, which is only expressed in adult SCN5A isoforms. Because the adult SCN5A isoform was poorly expressed, without functional differences between R222Q and control in both embryoid bodies and cell sheet preparations (cultured for 29-35 days), we created heart-on-a-chip biowires which promote myocardial maturation. Indeed, biowires expressed primarily adult SCN5A with R222Q preparations displaying (arrhythmogenic) short action potentials, altered Na⁺ channel biophysical properties and lower contractility compared to corrected controls. Comprehensive RNA sequencing revealed differential gene regulation between R222Q and control biowires in cellular pathways related to sarcoplasmic reticulum and dystroglycan complex as well as biological processes related to calcium ion regulation and action potential. Additionally, R222Q biowires had marked reductions in actin expression accompanied by profound sarcoplasmic disarray, without differences in cell composition (fibroblast, endothelial cells, and cardiomyocytes) compared to corrected biowires. In conclusion, we demonstrate that in addition to altering cardiac electrophysiology and Na⁺ current, the R222Q mutation also causes profound sarcomere disruptions and mechanical destabilization. Possible mechanisms for these observations are discussed.

Keywords: Arrhythmias; Cardiac tissue; Dilated cardiomyopathy; Mutation; Nav1.5; SCN5A; Sodium channel; iPSC.

Publication types

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

MeSH terms

Adult
Cardiomyopathy, Dilated* / genetics
Endothelial Cells
Humans
Induced Pluripotent Stem Cells*
Lab-On-A-Chip Devices
Myocytes, Cardiac

Substances

SCN5A protein, human

Abstract

Publication types

MeSH terms

Substances

Grants and funding