Targeted next-generation sequencing helps to decipher the genetic and phenotypic heterogeneity of hypertrophic cardiomyopathy

Cecconi,Massimiliano; Parodi,Maria I.; Formisano,Francesco; Spirito,Paolo; Autore,Camillo; Musumeci,Maria B.; Favale,Stefano; Forleo,Cinzia; Rapezzi,Claudio; Biagini,Elena; Davì,Sabrina; Canepa,Elisabetta; Pennese,Loredana; Castagnetta,Mauro; Degiorgio,Dario; Coviello,Domenico A.

doi:10.3892/ijmm.2016.2732

Targeted next-generation sequencing helps to decipher the genetic and phenotypic heterogeneity of hypertrophic cardiomyopathy

Authors:
- Massimiliano Cecconi
- Maria I. Parodi
- Francesco Formisano
- Paolo Spirito
- Camillo Autore
- Maria B. Musumeci
- Stefano Favale
- Cinzia Forleo
- Claudio Rapezzi
- Elena Biagini
- Sabrina Davì
- Elisabetta Canepa
- Loredana Pennese
- Mauro Castagnetta
- Dario Degiorgio
- Domenico A. Coviello
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Affiliations: Laboratory of Human Genetics, E.O. Ospedali Galliera, Genova, Italy, Cardiology Unit, E.O. Ospedali Galliera, Genova, Italy, Cardiology Unit, St. Andrea Hospital, Sapienza University, Rome, Italy, Cardiology Unit, Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy, Department of Experimental, Diagnostic and Specialty Medicine, S. Orsola Hospital, University of Bologna, Bologna, Italy
Published online on: September 7, 2016 https://doi.org/10.3892/ijmm.2016.2732
Pages: 1111-1124
Copyright: © Cecconi et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Hypertrophic cardiomyopathy (HCM) is mainly associated with myosin, heavy chain 7 (MYH7) and myosin binding protein C, cardiac (MYBPC3) mutations. In order to better explain the clinical and genetic heterogeneity in HCM patients, in this study, we implemented a target-next generation sequencing (NGS) assay. An Ion AmpliSeq™ Custom Panel for the enrichment of 19 genes, of which 9 of these did not encode thick/intermediate and thin myofilament (TTm) proteins and, among them, 3 responsible of HCM phenocopy, was created. Ninety-two DNA samples were analyzed by the Ion Personal Genome Machine: 73 DNA samples (training set), previously genotyped in some of the genes by Sanger sequencing, were used to optimize the NGS strategy, whereas 19 DNA samples (discovery set) allowed the evaluation of NGS performance. In the training set, we identified 72 out of 73 expected mutations and 15 additional mutations: the molecular diagnosis was achieved in one patient with a previously wild-type status and the pre-excitation syndrome was explained in another. In the discovery set, we identified 20 mutations, 5 of which were in genes encoding non-TTm proteins, increasing the diagnostic yield by approximately 20%: a single mutation in genes encoding non-TTm proteins was identified in 2 out of 3 borderline HCM patients, whereas co-occuring mutations in genes encoding TTm and galactosidase alpha (GLA) altered proteins were characterized in a male with HCM and multiorgan dysfunction. Our combined targeted NGS-Sanger sequencing-based strategy allowed the molecular diagnosis of HCM with greater efficiency than using the conventional (Sanger) sequencing alone. Mutant alleles encoding non-TTm proteins may aid in the complete understanding of the genetic and phenotypic heterogeneity of HCM: co-occuring mutations of genes encoding TTm and non-TTm proteins could explain the wide variability of the HCM phenotype, whereas mutations in genes encoding only the non-TTm proteins are identifiable in patients with a milder HCM status.

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