Multilevel analyses of SCN5A mutations in arrhythmogenic right ventricular dysplasia/cardiomyopathy suggest non-canonical mechanisms for disease pathogenesis

Anneline S.J.M. Te Riele, Esperanza Agullo-Pascual, Cynthia A. James, Alejandra Leo-Macias, Marina Cerrone, Mingliang Zhang, Xianming Lin, Bin Lin, Eli Rothenberg, Nara L. Sobreira, Nuria Amat-Alarcon, Roos F. Marsman, Brittney Murray, Crystal Tichnell, Jeroen F. Van Der Heijden, Dennis Dooijes, Toon A.B. Van Veen, Harikrishna Tandri, Steven J. Fowler, Richard N.W. Hauer & 13 others Gordon F. Tomaselli, Maarten P. Van Den Berg, Matthew R.G. Taylor, Francesca Brun, Gianfranco Sinagra, Arthur A.M. Wilde, Luisa Mestroni, Connie R. Bezzina, Hugh Calkins, J. Peter Van Tintelen, Lei Bu, Mario Delmar, Daniel P. Judge

Research output: Contribution to journalArticle

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Abstract

Aims 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. Methods and results 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. Conclusions 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.

Original languageEnglish (US)
Pages (from-to)102-111
Number of pages10
JournalCardiovascular Research
Volume113
Issue number1
DOIs
StatePublished - Jan 1 2017
Externally publishedYes

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Arrhythmogenic Right Ventricular Dysplasia
Multilevel Analysis
Mutation
Cadherins
Clustered Regularly Interspaced Short Palindromic Repeats
Sodium
Exome
Induced Pluripotent Stem Cells
Desmosomes
Sodium Channels
Cardiomyopathies
Fluorescence Microscopy
Cardiac Myocytes
Constriction
Blood Cells
Technology
Genes
Proteins

Keywords

  • Arrhythmogenic right ventricular cardiomyopathy
  • Cardiomyopathy
  • Genetics
  • Ion channel electrophysiology
  • SCN5A

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)

Cite this

Te Riele, A. S. J. M., Agullo-Pascual, E., James, C. A., Leo-Macias, A., Cerrone, M., Zhang, M., ... Judge, D. P. (2017). Multilevel analyses of SCN5A mutations in arrhythmogenic right ventricular dysplasia/cardiomyopathy suggest non-canonical mechanisms for disease pathogenesis. Cardiovascular Research, 113(1), 102-111. https://doi.org/10.1093/cvr/cvw234

Multilevel analyses of SCN5A mutations in arrhythmogenic right ventricular dysplasia/cardiomyopathy suggest non-canonical mechanisms for disease pathogenesis. / Te Riele, Anneline S.J.M.; Agullo-Pascual, Esperanza; James, Cynthia A.; Leo-Macias, Alejandra; Cerrone, Marina; Zhang, Mingliang; Lin, Xianming; Lin, Bin; Rothenberg, Eli; Sobreira, Nara L.; Amat-Alarcon, Nuria; Marsman, Roos F.; Murray, Brittney; Tichnell, Crystal; Van Der Heijden, Jeroen F.; Dooijes, Dennis; Van Veen, Toon A.B.; Tandri, Harikrishna; Fowler, Steven J.; Hauer, Richard N.W.; Tomaselli, Gordon F.; Van Den Berg, Maarten P.; Taylor, Matthew R.G.; Brun, Francesca; Sinagra, Gianfranco; Wilde, Arthur A.M.; Mestroni, Luisa; Bezzina, Connie R.; Calkins, Hugh; Van Tintelen, J. Peter; Bu, Lei; Delmar, Mario; Judge, Daniel P.

In: Cardiovascular Research, Vol. 113, No. 1, 01.01.2017, p. 102-111.

Research output: Contribution to journalArticle

Te Riele, ASJM, Agullo-Pascual, E, James, CA, Leo-Macias, A, Cerrone, M, Zhang, M, Lin, X, Lin, B, Rothenberg, E, Sobreira, NL, Amat-Alarcon, N, Marsman, RF, Murray, B, Tichnell, C, Van Der Heijden, JF, Dooijes, D, Van Veen, TAB, Tandri, H, Fowler, SJ, Hauer, RNW, Tomaselli, GF, Van Den Berg, MP, Taylor, MRG, Brun, F, Sinagra, G, Wilde, AAM, Mestroni, L, Bezzina, CR, Calkins, H, Van Tintelen, JP, Bu, L, Delmar, M & Judge, DP 2017, 'Multilevel analyses of SCN5A mutations in arrhythmogenic right ventricular dysplasia/cardiomyopathy suggest non-canonical mechanisms for disease pathogenesis', Cardiovascular Research, vol. 113, no. 1, pp. 102-111. https://doi.org/10.1093/cvr/cvw234
Te Riele, Anneline S.J.M. ; Agullo-Pascual, Esperanza ; James, Cynthia A. ; Leo-Macias, Alejandra ; Cerrone, Marina ; Zhang, Mingliang ; Lin, Xianming ; Lin, Bin ; Rothenberg, Eli ; Sobreira, Nara L. ; Amat-Alarcon, Nuria ; Marsman, Roos F. ; Murray, Brittney ; Tichnell, Crystal ; Van Der Heijden, Jeroen F. ; Dooijes, Dennis ; Van Veen, Toon A.B. ; Tandri, Harikrishna ; Fowler, Steven J. ; Hauer, Richard N.W. ; Tomaselli, Gordon F. ; Van Den Berg, Maarten P. ; Taylor, Matthew R.G. ; Brun, Francesca ; Sinagra, Gianfranco ; Wilde, Arthur A.M. ; Mestroni, Luisa ; Bezzina, Connie R. ; Calkins, Hugh ; Van Tintelen, J. Peter ; Bu, Lei ; Delmar, Mario ; Judge, Daniel P. / Multilevel analyses of SCN5A mutations in arrhythmogenic right ventricular dysplasia/cardiomyopathy suggest non-canonical mechanisms for disease pathogenesis. In: Cardiovascular Research. 2017 ; Vol. 113, No. 1. pp. 102-111.
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title = "Multilevel analyses of SCN5A mutations in arrhythmogenic right ventricular dysplasia/cardiomyopathy suggest non-canonical mechanisms for disease pathogenesis",
abstract = "Aims 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. Methods and results 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. Conclusions 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.",
keywords = "Arrhythmogenic right ventricular cardiomyopathy, Cardiomyopathy, Genetics, Ion channel electrophysiology, SCN5A",
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TY - JOUR

T1 - Multilevel analyses of SCN5A mutations in arrhythmogenic right ventricular dysplasia/cardiomyopathy suggest non-canonical mechanisms for disease pathogenesis

AU - Te Riele, Anneline S.J.M.

AU - Agullo-Pascual, Esperanza

AU - James, Cynthia A.

AU - Leo-Macias, Alejandra

AU - Cerrone, Marina

AU - Zhang, Mingliang

AU - Lin, Xianming

AU - Lin, Bin

AU - Rothenberg, Eli

AU - Sobreira, Nara L.

AU - Amat-Alarcon, Nuria

AU - Marsman, Roos F.

AU - Murray, Brittney

AU - Tichnell, Crystal

AU - Van Der Heijden, Jeroen F.

AU - Dooijes, Dennis

AU - Van Veen, Toon A.B.

AU - Tandri, Harikrishna

AU - Fowler, Steven J.

AU - Hauer, Richard N.W.

AU - Tomaselli, Gordon F.

AU - Van Den Berg, Maarten P.

AU - Taylor, Matthew R.G.

AU - Brun, Francesca

AU - Sinagra, Gianfranco

AU - Wilde, Arthur A.M.

AU - Mestroni, Luisa

AU - Bezzina, Connie R.

AU - Calkins, Hugh

AU - Van Tintelen, J. Peter

AU - Bu, Lei

AU - Delmar, Mario

AU - Judge, Daniel P.

PY - 2017/1/1

Y1 - 2017/1/1

N2 - Aims 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. Methods and results 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. Conclusions 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.

AB - Aims 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. Methods and results 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. Conclusions 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.

KW - Arrhythmogenic right ventricular cardiomyopathy

KW - Cardiomyopathy

KW - Genetics

KW - Ion channel electrophysiology

KW - SCN5A

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U2 - 10.1093/cvr/cvw234

DO - 10.1093/cvr/cvw234

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SP - 102

EP - 111

JO - Cardiovascular Research

JF - Cardiovascular Research

SN - 0008-6363

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