title: Adeno-associated virus-mediated CASQ2 delivery rescues phenotypic alterations in a patient-specific model of recessive catecholaminergic polymorphic ventricular tachycardia.
abstract: Catecholaminergic Polymorphic Ventricular Tachycardia type 2 (CPVT2) is a highly lethal recessive arrhythmogenic disease caused by mutations in the calsequestrin-2 (CASQ2) gene. We have previously demonstrated that viral transfer of the wild-type (WT) CASQ2 gene prevents the development of CPVT2 in a genetically induced mouse model of the disease homozygous carrier of the R33Q mutation. In the present study, we investigated the efficacy of the virally mediated gene therapy in cardiomyocytes (CMs) differentiated from induced pluripotent stem cells (iPSCs) obtained from a patient carrying the homozygous CASQ2-G112+5X mutation. To this end, we infected cells with an Adeno-Associated Viral vector serotype 9 (AAV9) encoding the human CASQ2 gene (AAV9-hCASQ2). Administration of the human WT CASQ2 gene was capable and sufficient to restore the physiological expression of calsequestrin-2 protein and to rescue functional defects of the patient-specific iPSC-derived CMs. Indeed, after viral gene transfer, we observed a remarkable decrease in the percentage of delayed afterdepolarizations (DADs) developed by the diseased CMs upon adrenergic stimulation, the calcium transient amplitude was re-established and the density and duration of calcium sparks were normalized. We therefore demonstrate the efficacy of the AAV9-mediated gene replacement therapy for CPVT2 in a human cardiac-specific model system, supporting the view that the gene-therapy tested is curative in models with different human mutations of CPVT.
authors.affiliation: Molecular Cardiology, IRCCS Fondazione Salvatore Maugeri, Pavia, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Lodola
authors.affiliation: Laboratory of Inflammation and Immunology in Cardiovascular Pathologies, Humanitas Research Hospital, Rozzano (MI), Italy.
authors.forename: Diego
authors.initials: D
authors.lastname: Morone
authors.affiliation: Molecular Cardiology, IRCCS Fondazione Salvatore Maugeri, Pavia, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Denegri
authors.affiliation: Molecular Cardiology, IRCCS Fondazione Salvatore Maugeri, Pavia, Italy.
authors.forename: Rossana
authors.initials: R
authors.lastname: Bongianino
authors.affiliation: Laboratory of Inflammation and Immunology in Cardiovascular Pathologies, Humanitas Research Hospital, Rozzano (MI), Italy.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Nakahama
authors.affiliation: Laboratory of Inflammation and Immunology in Cardiovascular Pathologies, Humanitas Research Hospital, Rozzano (MI), Italy.|Institute of Genetic and Biomedical Research (IRGB) - UOS Milan, National Research Council of Italy, Milan, Italy.
authors.forename: Lucia
authors.initials: L
authors.lastname: Rutigliano
authors.affiliation: Laboratory of Inflammation and Immunology in Cardiovascular Pathologies, Humanitas Research Hospital, Rozzano (MI), Italy.
authors.forename: Rosanna
authors.initials: R
authors.lastname: Gosetti
authors.affiliation: Molecular Cardiology, IRCCS Fondazione Salvatore Maugeri, Pavia, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Rizzo
authors.affiliation: Molecular Cardiology, IRCCS Fondazione Salvatore Maugeri, Pavia, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Vollero
authors.affiliation: Unit of Clinical Neurophysiology & Neurodiagnostic Skin Biopsy, IRCCS Fondazione Salvatore Maugeri, Pavia, Italy.
authors.forename: Michelangelo
authors.initials: M
authors.lastname: Buonocore
authors.affiliation: Molecular Cardiology, IRCCS Fondazione Salvatore Maugeri, Pavia, Italy.
authors.forename: Carlo
authors.initials: C
authors.lastname: Napolitano
authors.affiliation: Laboratory of Inflammation and Immunology in Cardiovascular Pathologies, Humanitas Research Hospital, Rozzano (MI), Italy.|Institute of Genetic and Biomedical Research (IRGB) - UOS Milan, National Research Council of Italy, Milan, Italy.|Humanitas University, Rozzano (MI), Italy.
authors.forename: Gianluigi
authors.identifier: 0000-0003-0481-6843
authors.initials: G
authors.lastname: Condorelli
authors.affiliation: Molecular Cardiology, IRCCS Fondazione Salvatore Maugeri, Pavia, Italy.|Department of Molecular Medicine, University of Pavia, Pavia, Italy.
authors.forename: Silvia G
authors.identifier: 0000-0001-6877-0288
authors.initials: SG
authors.lastname: Priori
authors.affiliation: Laboratory of Inflammation and Immunology in Cardiovascular Pathologies, Humanitas Research Hospital, Rozzano (MI), Italy.|Institute of Genetic and Biomedical Research (IRGB) - UOS Milan, National Research Council of Italy, Milan, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Di Pasquale
mesh_terms: D001145:Arrhythmias, Cardiac / Q000378:metabolism / Q000473:pathology; D001706:Biopsy; D002118:Calcium / Q000378:metabolism; D002155:Calsequestrin / Q000235:genetics*; D002395:Catecholamines / Q000378:metabolism*; D002454:Cell Differentiation; D000229:Dependovirus / Q000378:metabolism*; D005260:Female; D018014:Gene Transfer Techniques; D005808:Genes, Recessive; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism; D008297:Male; D008954:Models, Biological; D032383:Myocytes, Cardiac / Q000378:metabolism / Q000473:pathology; D010375:Pedigree; D010641:Phenotype; D012867:Skin / Q000473:pathology; D017180:Tachycardia, Ventricular / Q000473:pathology / Q000503:physiopathology / Q000628:therapy*; D000098850:Polymorphic Catecholaminergic Ventricular Tachycardia
chemical_list: C443109:CASQ2 protein, human; D002155:Calsequestrin; D002395:Catecholamines; D002118:Calcium