| Title | Modeling polymorphic ventricular tachycardia at rest using patient-specific induced pluripotent stem cell-derived cardiomyocytes. |
| Publication Type | Journal Article |
| Year of Publication | 2020 |
| Authors | Sleiman Y, Souidi M, Kumar R, Yang E, Jaffré F, Zhou T, Bernardin A, Reiken S, Cazorla O, Kajava AV, Moreau A, Pasquié J-L, Marks AR, Lerman BB, Chen S, Cheung JW, Evans T, Lacampagne A, Meli AC |
| Journal | EBioMedicine |
| Volume | 60 |
| Pagination | 103024 |
| Date Published | 2020 Oct |
| ISSN | 2352-3964 |
| Keywords | Alleles, Calcium, Calcium Signaling, Cell Differentiation, CRISPR-Cas Systems, Genotype, Homeostasis, Humans, Immunohistochemistry, Induced Pluripotent Stem Cells, Models, Biological, Mutation, Myocytes, Cardiac, Protein Processing, Post-Translational, Stem Cell Transplantation, Tachycardia, Ventricular |
| Abstract | <p><b>BACKGROUND: </b>While mutations in the cardiac type 2 ryanodine receptor (RyR2) have been linked to exercise-induced or catecholaminergic polymorphic ventricular tachycardia (CPVT), its association with polymorphic ventricular tachycardia (PMVT) occurring at rest is unclear. We aimed at constructing a patient-specific human-induced pluripotent stem cell (hiPSC) model of PMVT occurring at rest linked to a single point mutation in RyR2.</p><p><b>METHODS: </b>Blood samples were obtained from a patient with PMVT at rest due to a heterozygous RyR2-H29D mutation. Patient-specific hiPSCs were generated from the blood samples, and the hiPSC-derived cardiomyocytes (CMs) were generated via directed differentiation. Using CRIPSR/Cas9 technology, isogenic controls were generated by correcting the RyR2-H29D mutation. Using patch-clamp, fluorescent confocal microscopy and video-image-based analysis, the molecular and functional properties of RyR2-H29D hiPSCCMs and control hiPSCCMs were compared.</p><p><b>FINDINGS: </b>RyR2-H29D hiPSCCMs exhibit intracellular sarcoplasmic reticulum (SR) Ca leak through RyR2 under physiological pacing. RyR2-H29D enhances the contribution of inositol 1,4,5-trisphosphate receptors to excitation-contraction coupling (ECC) that exacerbates abnormal Ca release in RyR2-H29D hiPSCCMs. RyR2-H29D hiPSCCMs exhibit shorter action potentials, delayed afterdepolarizations, arrhythmias and aberrant contractile properties compared to isogenic controls. The RyR2-H29D mutation causes post-translational remodeling that is fully reversed with isogenic controls.</p><p><b>INTERPRETATION: </b>To conclude, in a model based on a RyR2 point mutation that is associated with short-coupled PMVT at rest, RyR2-H29D hiPSCCMs exhibited aberrant intracellular Ca homeostasis, shortened action potentials, arrhythmias and abnormal contractile properties.</p><p><b>FUNDING: </b>French Muscular Dystrophy Association (AFM; project 16,073, MNM2 2012 and 20,225), "Fondation de la Recherche Médicale" (FRM; SPF20130526710), "Institut National pour la Santé et la Recherche Médicale" (INSERM), National Institutes of Health (ARM; R01 HL145473) and New York State Department of Health (NYSTEM C029156).</p> |
| DOI | 10.1016/j.ebiom.2020.103024 |
| Alternate Journal | EBioMedicine |
| PubMed ID | 32980690 |
| PubMed Central ID | PMC7519379 |