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J Cell Mol Med. 2019 Mar;23(3):2125-2135. doi: 10.1111/jcmm.14124. Epub 2019 Jan 8.

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

Author information

1
Department of Physiology, Biophysics and Systems Biology, Rappaport Faculty of Medicine, Technion - Israel Institute of Technology, Haifa, Israel.
2
Department of Biotechnology, Technion - Israel Institute of Technology, Haifa, Israel.
3
Department of Molecular and Integrative Physiology, University of Michigan, Ann Arbor, Michigan.
4
Leviev Heart Center, Sheba Medical Center, Ramat Gan, Israel.
5
Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel.
6
Victor Babes National Institute of Pathology, Bucharest, Romania.
7
Health in Code, Corunna, Spain.
8
Department of Hematology, Hemostasis, Oncology and Stem Cell Transplantation, Hannover Medical School, Hannover, Germany.
9
Department of Cardiology and Angiology, Hannover Medical School, Hannover, Germany.

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 If density, and DMD female and male iPSC-CMs showed increased ICa,L density. Our findings demonstrate cellular mechanisms underlying electrophysiological abnormalities and cardiac arrhythmias in DMD.

KEYWORDS:

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

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