Modeling polymorphic ventricular tachycardia at rest using patient-specific induced pluripotent stem cell-derived cardiomyocytes

Yvonne Sleiman (1) , Monia Souidi (1) , Ritu Kumar (2) , Ellen Yang (2) , Fabrice Jaffré (2) , Ting Zhou (2) , Albin Bernardin (1) , Steve Reiken (3) , Olivier Cazorla (1) , Andrey Kajava (4) , Adrien Moreau , Jean Luc Pasquié (1) , Andrew Marks (3) , Bruce Lerman (2) , Shuibing Chen (2) , Jim Cheung (2) , Todd Evans (2) , Alain Lacampagne (1) , Albano C Meli (1)

1 PhyMedExp - Physiologie & médecine expérimentale du Cœur et des Muscles [U 1046]
2 Weill Medical College of Cornell University [New York]
3 Columbia University College of Physicians and Surgeons
4 CRBM - Centre de recherche en Biologie cellulaire de Montpellier

Background: 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.Methods: 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 hiPSCsingle bondCMs and control hiPSCsingle bondCMs were compared.Findings: RyR2-H29D hiPSCsingle bondCMs exhibit intracellular sarcoplasmic reticulum (SR) Ca2+ 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 Ca2+ release in RyR2-H29D hiPSCsingle bondCMs. RyR2-H29D hiPSCsingle bondCMs 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.Interpretation: To conclude, in a model based on a RyR2 point mutation that is associated with short-coupled PMVT at rest, RyR2-H29D hiPSCsingle bondCMs exhibited aberrant intracellular Ca2+ homeostasis, shortened action potentials, arrhythmias and abnormal contractile properties.

Mots clés: PMVT; Ryanodine recepto; r hiPSC-derived cardiomyocytes; Calcium handling; Contractile properties

Domaines: Cardiologie et système cardiovasculaire

Yvonne Sleiman, Monia Souidi, Ritu Kumar, Ellen Yang, Fabrice Jaffré, et al.. Modeling polymorphic ventricular tachycardia at rest using patient-specific induced pluripotent stem cell-derived cardiomyocytes. EBioMedicine, 2020, 60, pp.103024. ⟨10.1016/j.ebiom.2020.103024⟩. ⟨hal-03089303v2⟩