title: Multilevel analyses of SCN5A mutations in arrhythmogenic right ventricular dysplasia/cardiomyopathy suggest non-canonical mechanisms for disease pathogenesis.
abstract: Arrhythmogenic Right Ventricular Dysplasia/Cardiomyopathy (ARVD/C) is often associated with desmosomal mutations. Recent studies suggest an interaction between the desmosome and sodium channel protein Nav1.5. We aimed to determine the prevalence and biophysical properties of mutations in SCN5A (the gene encoding Nav1.5) in ARVD/C.
  
  
  
  We performed whole-exome sequencing in six ARVD/C patients (33% male, 38.2 ± 12.1 years) without a desmosomal mutation. We found a rare missense variant (p.Arg1898His; R1898H) in SCN5A in one patient. We generated induced pluripotent stem cell-derived cardiomyocytes (hIPSC-CMs) from the patient's peripheral blood mononuclear cells. The variant was then corrected (R1898R) using Clustered Regularly Interspaced Short Palindromic Repeats/Cas9 technology, allowing us to study the impact of the R1898H substitution in the same cellular background. Whole-cell patch clamping revealed a 36% reduction in peak sodium current (P = 0.002); super-resolution fluorescence microscopy showed reduced abundance of NaV1.5 (P = 0.005) and N-Cadherin (P = 0.026) clusters at the intercalated disc. Subsequently, we sequenced SCN5A in an additional 281 ARVD/C patients (60% male, 34.8 ± 13.7 years, 52% desmosomal mutation-carriers). Five (1.8%) subjects harboured a putatively pathogenic SCN5A variant (p.Tyr416Cys, p.Leu729del, p.Arg1623Ter, p.Ser1787Asn, and p.Val2016Met). SCN5A variants were associated with prolonged QRS duration (119 ± 15 vs. 94 ± 14 ms, P < 0.01) and all SCN5A variant carriers had major structural abnormalities on cardiac imaging.
  
  
  
  Almost 2% of ARVD/C patients harbour rare SCN5A variants. For one of these variants, we demonstrated reduced sodium current, Nav1.5 and N-Cadherin clusters at junctional sites. This suggests that Nav1.5 is in a functional complex with adhesion molecules, and reveals potential non-canonical mechanisms by which Nav1.5 dysfunction causes cardiomyopathy.
authors.affiliation: Department of Medicine, Division of Cardiology, Johns Hopkins University School of Medicine, 1800 Orleans Street, Baltimore, MD, USA.|Division of Cardiology, University Medical Center Utrecht, Heidelberglaan 100, Utrecht, the Netherlands.|Netherlands Heart Institute, Moreelsepark 1, Utrecht, the Netherlands.
authors.forename: Anneline S J M
authors.initials: AS
authors.lastname: Te Riele
authors.affiliation: Leon H. Charney Division of Cardiology, New York University School of Medicine, 550 First Avenue, New York, NY, USA.
authors.forename: Esperanza
authors.initials: E
authors.lastname: Agullo-Pascual
authors.affiliation: Department of Medicine, Division of Cardiology, Johns Hopkins University School of Medicine, 1800 Orleans Street, Baltimore, MD, USA.
authors.forename: Cynthia A
authors.initials: CA
authors.lastname: James
authors.affiliation: Leon H. Charney Division of Cardiology, New York University School of Medicine, 550 First Avenue, New York, NY, USA.
authors.forename: Alejandra
authors.initials: A
authors.lastname: Leo-Macias
authors.affiliation: Leon H. Charney Division of Cardiology, New York University School of Medicine, 550 First Avenue, New York, NY, USA.
authors.forename: Marina
authors.initials: M
authors.lastname: Cerrone
authors.affiliation: Leon H. Charney Division of Cardiology, New York University School of Medicine, 550 First Avenue, New York, NY, USA.
authors.forename: Mingliang
authors.initials: M
authors.lastname: Zhang
authors.affiliation: Leon H. Charney Division of Cardiology, New York University School of Medicine, 550 First Avenue, New York, NY, USA.
authors.forename: Xianming
authors.initials: X
authors.lastname: Lin
authors.affiliation: Leon H. Charney Division of Cardiology, New York University School of Medicine, 550 First Avenue, New York, NY, USA.
authors.forename: Bin
authors.initials: B
authors.lastname: Lin
authors.affiliation: McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine, 720 Rutland Avenue, Baltimore, MD, USA.
authors.forename: Nara L
authors.initials: NL
authors.lastname: Sobreira
authors.affiliation: Department of Medicine, Division of Cardiology, Johns Hopkins University School of Medicine, 1800 Orleans Street, Baltimore, MD, USA.
authors.forename: Nuria
authors.initials: N
authors.lastname: Amat-Alarcon
authors.affiliation: Heart Centre, Department of Clinical and Experimental Cardiology, Academic Medical Center, Meibergdreef 9, Amsterdam, the Netherlands.
authors.forename: Roos F
authors.initials: RF
authors.lastname: Marsman
authors.affiliation: Department of Medicine, Division of Cardiology, Johns Hopkins University School of Medicine, 1800 Orleans Street, Baltimore, MD, USA.
authors.forename: Brittney
authors.initials: B
authors.lastname: Murray
authors.affiliation: Department of Medicine, Division of Cardiology, Johns Hopkins University School of Medicine, 1800 Orleans Street, Baltimore, MD, USA.
authors.forename: Crystal
authors.initials: C
authors.lastname: Tichnell
authors.affiliation: Division of Cardiology, University Medical Center Utrecht, Heidelberglaan 100, Utrecht, the Netherlands.
authors.forename: Jeroen F
authors.initials: JF
authors.lastname: van der Heijden
authors.affiliation: Department of Medical Genetics, University Medical Center Utrecht, Heidelberglaan 100, Utrecht, the Netherlands.
authors.forename: Dennis
authors.initials: D
authors.lastname: Dooijes
authors.affiliation: Department of Medical Physiology, Division of Heart and Lungs, University Medical Center Utrecht, Yalelaan 50, Utrecht, the Netherlands.
authors.forename: Toon A B
authors.initials: TA
authors.lastname: van Veen
authors.affiliation: Department of Medicine, Division of Cardiology, Johns Hopkins University School of Medicine, 1800 Orleans Street, Baltimore, MD, USA.
authors.forename: Harikrishna
authors.initials: H
authors.lastname: Tandri
authors.affiliation: Leon H. Charney Division of Cardiology, New York University School of Medicine, 550 First Avenue, New York, NY, USA.
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Fowler
authors.affiliation: Division of Cardiology, University Medical Center Utrecht, Heidelberglaan 100, Utrecht, the Netherlands.|Netherlands Heart Institute, Moreelsepark 1, Utrecht, the Netherlands.
authors.forename: Richard N W
authors.initials: RN
authors.lastname: Hauer
authors.affiliation: Department of Medicine, Division of Cardiology, Johns Hopkins University School of Medicine, 1800 Orleans Street, Baltimore, MD, USA.
authors.forename: Gordon
authors.initials: G
authors.lastname: Tomaselli
authors.affiliation: Department of Cardiology, University Medical Center Groningen, University of Groningen, Hanzeplein 1, Groningen, the Netherlands.
authors.forename: Maarten P
authors.initials: MP
authors.lastname: van den Berg
authors.affiliation: Cardiovascular Institute and Adult Medical Genetics, University of Colorado Denver, 12605 E 16th Avenue, Aurora, CO, USA.
authors.forename: Matthew R G
authors.initials: MR
authors.lastname: Taylor
authors.affiliation: Cardiovascular Department, Ospedali Riuniti and University of Trieste, Via Farneto 3, Trieste, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Brun
authors.affiliation: Cardiovascular Department, Ospedali Riuniti and University of Trieste, Via Farneto 3, Trieste, Italy.
authors.forename: Gianfranco
authors.initials: G
authors.lastname: Sinagra
authors.affiliation: Heart Centre, Department of Clinical and Experimental Cardiology, Academic Medical Center, Meibergdreef 9, Amsterdam, the Netherlands.
authors.forename: Arthur A M
authors.initials: AA
authors.lastname: Wilde
authors.affiliation: Cardiovascular Institute and Adult Medical Genetics, University of Colorado Denver, 12605 E 16th Avenue, Aurora, CO, USA.
authors.forename: Luisa
authors.initials: L
authors.lastname: Mestroni
authors.affiliation: Heart Centre, Department of Clinical and Experimental Cardiology, Academic Medical Center, Meibergdreef 9, Amsterdam, the Netherlands.
authors.forename: Connie R
authors.initials: CR
authors.lastname: Bezzina
authors.affiliation: Department of Medicine, Division of Cardiology, Johns Hopkins University School of Medicine, 1800 Orleans Street, Baltimore, MD, USA.
authors.forename: Hugh
authors.initials: H
authors.lastname: Calkins
authors.affiliation: Department of Cardiology, University Medical Center Groningen, University of Groningen, Hanzeplein 1, Groningen, the Netherlands.|Department of Clinical Genetics, Academic Medical Center Amsterdam, University of Amsterdam, Meibergdreef 9, Amsterdam, the Netherlands.|Department of Genetics, University of Groningen, University Medical Center Groningen, Hanzeplein 1, Groningen, the Netherlands.
authors.forename: J
authors.initials: J
authors.lastname: Peter van Tintelen
authors.affiliation: Leon H. Charney Division of Cardiology, New York University School of Medicine, 550 First Avenue, New York, NY, USA.|Department of Medicine, Division of Cardiology, Johns Hopkins University School of Medicine, 1800 Orleans Street, Baltimore, MD, USA.
authors.forename: Lei
authors.initials: L
authors.lastname: Bu
authors.affiliation: Leon H. Charney Division of Cardiology, New York University School of Medicine, 550 First Avenue, New York, NY, USA.|Department of Medicine, Division of Cardiology, Johns Hopkins University School of Medicine, 1800 Orleans Street, Baltimore, MD, USA.
authors.forename: Mario
authors.initials: M
authors.lastname: Delmar
authors.affiliation: Department of Medicine, Division of Cardiology, Johns Hopkins University School of Medicine, 1800 Orleans Street, Baltimore, MD, USA; djudge@jhmi.edu mario.delmar@nyumc.org lei.bu@med.nyu.edu.
authors.forename: Daniel P
authors.initials: DP
authors.lastname: Judge
mesh_terms: D000328:Adult; D015703:Antigens, CD / Q000378:metabolism; D019571:Arrhythmogenic Right Ventricular Dysplasia / Q000000981:diagnostic imaging / Q000235:genetics* / Q000378:metabolism; D064113:CRISPR-Cas Systems; D015820:Cadherins / Q000378:metabolism; D002454:Cell Differentiation; D004252:DNA Mutational Analysis; D004562:Electrocardiography; D059472:Exome; D005260:Female; D000072669:Gene Editing / Q000379:methods; D005787:Gene Frequency; D020022:Genetic Predisposition to Disease; D055106:Genome-Wide Association Study; D057809:HEK293 Cells; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism; D008279:Magnetic Resonance Imaging; D008297:Male; D008564:Membrane Potentials; D008875:Middle Aged; D055361:Multilevel Analysis; D020125:Mutation, Missense; D032383:Myocytes, Cardiac / Q000378:metabolism; D062554:NAV1.5 Voltage-Gated Sodium Channel / Q000235:genetics* / Q000378:metabolism; D009426:Netherlands; D010641:Phenotype; D012964:Sodium / Q000378:metabolism; D014162:Transfection; D014481:United States; D055815:Young Adult
chemical_list: D015703:Antigens, CD; C050361:CDH2 protein, human; D015820:Cadherins; D062554:NAV1.5 Voltage-Gated Sodium Channel; C568320:SCN5A protein, human; D012964:Sodium
keywords: Arrhythmogenic right ventricular cardiomyopathy ; Cardiomyopathy ; Genetics ; Ion channel electrophysiology; SCN5A 