Journal Article

Contribution of exome sequencing for genetic diagnostic in arrhythmogenic right ventricular cardiomyopathy/dysplasia

Charron, Philippe, Donal, Erwan, Duthoit, Guillaume, Gandjbakhch, Estelle, Keren, Boris, Fressart, Veronique, Surget, Elodie, Hidden-Lucet, Françoise, Fedida, Joel, Richard, Pascale, Hery, Tiphaine, Villard, Eric

2017

Overview

Arrhythmogenic Right Ventricular Cardiomyopathy/Dysplasia (ARVC/D) is an inherited cardiomyopathy mainly caused by heterozygous desmosomal gene mutations, the major gene being PKP2. The genetic cause remains unknown in ~50% of probands with routine desmosomal gene screening. The aim of this study was to assess the diagnostic accuracy of whole exome sequencing (WES) in ARVC/D with negative genetic testing. WES was performed in 22 patients, all without a mutation identified in desmosomal genes. Putative pathogenic variants were screened in 96 candidate genes associated with other cardiomyopathies/channelopathies. The sequencing coverage depth of PKP2, DSP, DSG2, DSC2, JUP and TMEM43 exons was compared to the mean coverage distribution to detect large insertions/deletions. All suspected deletions were verified by real-time qPCR, Multiplex-Ligation-dependent-Probe-Amplification (MLPA) and cGH-Array. MLPA was performed in 50 additional gene-negative probands. Coverage-depth analysis from the 22 WES data identified two large heterozygous PKP2 deletions: one from exon 1 to 14 and one restricted to exon 4, confirmed by qPCR and MLPA. MLPA identified 2 additional PKP2 deletions (exon 1-7 and exon 1-14) in 50 additional probands confirming a significant frequency of large PKP2 deletions (5.7%) in gene-negative ARVC/D. Putative pathogenic heterozygous variants in EYA4, RBM20, PSEN1, and COX15 were identified in 4 unrelated probands. A rather high frequency (5.7%) of large PKP2 deletions, undetectable by Sanger sequencing, was detected as the cause of ARVC/D. Coverage-depth analysis through next-generation sequencing appears accurate to detect large deletions at the same time than conventional putative mutations in desmosomal and cardiomyopathy-associated genes.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Adolescent / Adult / Aged / Amplification / Analysis / Arrhythmogenic Right Ventricular Dysplasia - genetics / Biology and Life Sciences / Cardiology and cardiovascular system / Cardiomyopathy / Care and treatment / Comparative analysis / Diagnostic systems / DNA sequencing / Dysplasia / Electron Transport Complex IV - genetics / Exome / Exons / Female / Gene mutation / Gene Regulatory Networks / Gene sequencing / Genes / Genetic aspects / Genetic Predisposition to Disease / Genetic screening / Genetic testing / Genetics / Genome-Wide Association Study - methods / Genomics / Heart / Human genetics / Human health and pathology / Humans / Identification methods / Life Sciences / Male / Medical diagnosis / Medicine and Health Sciences / Middle Aged / Multiplexing / Mutation / Patients / Pedigree / Plakophilins - genetics / Presenilin-1 - genetics / Research and analysis methods / RNA-Binding Proteins - genetics / Sequence Analysis, DNA - methods / Sequence Deletion / Task forces / Test procedures / Trans-Activators - genetics / Ventricle / Young Adult