Electrophysiological abnormalities in induced pluripotent stem cell-derived cardiomyocytes generated from Duchenne muscular dystrophy patients

Binyamin Eisen, Ronen Ben Jehuda, Ashley J. Cuttitta, Lucy N. Mekies, Yuval Shemer, Polina Baskin, Irina Reiter, Lubna Willi, Dov Freimark, Mihaela Gherghiceanu, Lorenzo Monserrat, Michaela Scherr, Denise Hilfiker-Kleiner, Michael Arad, Daniel E. Michele*, Ofer Binah

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Duchenne muscular dystrophy (DMD) is an X-linked progressive muscle degenerative disease, caused by mutations in the dystrophin gene and resulting in death because of respiratory or cardiac failure. To investigate the cardiac cellular manifestation of DMD, we generated induced pluripotent stem cells (iPSCs) and iPSC-derived cardiomyocytes (iPSC-CMs) from two DMD patients: a male and female manifesting heterozygous carrier. Dystrophin mRNA and protein expression were analysed by qRT-PCR, RNAseq, Western blot and immunofluorescence staining. For comprehensive electrophysiological analysis, current and voltage clamp were used to record transmembrane action potentials and ion currents, respectively. Microelectrode array was used to record extracellular electrograms. X-inactive specific transcript (XIST) and dystrophin expression analyses revealed that female iPSCs underwent X chromosome reactivation (XCR) or erosion of X chromosome inactivation, which was maintained in female iPSC-CMs displaying mixed X chromosome expression of wild type (WT) and mutated alleles. Both DMD female and male iPSC-CMs presented low spontaneous firing rate, arrhythmias and prolonged action potential duration. DMD female iPSC-CMs displayed increased beat rate variability (BRV). DMD male iPSC-CMs manifested decreased I f density, and DMD female and male iPSC-CMs showed increased I Ca,L density. Our findings demonstrate cellular mechanisms underlying electrophysiological abnormalities and cardiac arrhythmias in DMD.

Original languageEnglish
Pages (from-to)2125-2135
Number of pages11
JournalJournal of Cellular and Molecular Medicine
Volume23
Issue number3
DOIs
StatePublished - Mar 2019

Funding

FundersFunder number
Association Duchenne Israel
US‐Israel Binational Sciences Foundation
National Institute of Arthritis and Musculoskeletal and Skin DiseasesR01AR068428
Bloom's Syndrome Foundation
American Dysautonomia Institute
University of Michigan
United States-Israel Binational Science Foundation
Israel Science Foundation

    Keywords

    • Duchenne muscular dystrophy
    • X chromosome inactivation
    • arrhythmia
    • dilated cardiomyopathy
    • induced pluripotent stem cell-derived cardiomyocytes

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