New Mutations in NEB Gene Discovered by Targeted Next-Generation Sequencing in Nemaline Myopathy Italian Patients

Daniela Piga, Francesca Magri, Dario Ronchi, Stefania Corti, Denise Cassandrini, Eugenio Mercuri, Giorgio Tasca, Enrico Bertini, Fabiana Fattori, Antonio Toscano, Sonia Messina, Isabella Moroni, Marina Mora, Maurizio Moggio, Irene Colombo, Teresa Giugliano, Marika Pane, Chiara Fiorillo, Adele D’Amico, Claudio Bruno & 3 others Vincenzo Nigro, Nereo Bresolin, Giacomo Pietro Comi

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Nemaline myopathy represents a group of clinically and genetically heterogeneous neuromuscular disorders. Different clinical-genetic entities have been characterized in the last few years, with implications for diagnostics and genetic counseling. Fifty percent of nemaline myopathy forms are due to NEB mutations, but genetic analysis of this large and complex gene by Sanger sequencing is time consuming and expensive. We selected 10 Italian patients with clinical and biopsy features suggestive for nemaline myopathy and negative for ACTA1, TPM2 and TPM3 mutations. We applied a targeted next-generation sequencing strategy designed to analyse NEB coding regions, the relative full introns and the promoter. We also evaluated copy number variations (by CGH array) and transcriptional changes by RNA Sanger sequencing, whenever possible. This combined strategy revealed 11 likely pathogenic variants in 8 of 10 patients. The molecular diagnosis was fully achieved in 3 of 8 patients, while only one heterozygous mutation was observed in 5 subjects. This approach revealed to be a fast and cost-effective way to analyse the large NEB gene in a small group of patients and might be promising for the detection of pathological variants of other genes featuring large coding regions and lacking mutational hotspots.

Original languageEnglish
Pages (from-to)351-359
Number of pages9
JournalJournal of Molecular Neuroscience
Volume59
Issue number3
DOIs
Publication statusPublished - 2016

Fingerprint

Nemaline Myopathies
Mutation
Genes
RNA Sequence Analysis
Genetic Counseling
Introns
Biopsy
Costs and Cost Analysis

Keywords

  • NEB mutations
  • Nemaline myopathy
  • Next-generation sequencing

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience

Cite this

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title = "New Mutations in NEB Gene Discovered by Targeted Next-Generation Sequencing in Nemaline Myopathy Italian Patients",
abstract = "Nemaline myopathy represents a group of clinically and genetically heterogeneous neuromuscular disorders. Different clinical-genetic entities have been characterized in the last few years, with implications for diagnostics and genetic counseling. Fifty percent of nemaline myopathy forms are due to NEB mutations, but genetic analysis of this large and complex gene by Sanger sequencing is time consuming and expensive. We selected 10 Italian patients with clinical and biopsy features suggestive for nemaline myopathy and negative for ACTA1, TPM2 and TPM3 mutations. We applied a targeted next-generation sequencing strategy designed to analyse NEB coding regions, the relative full introns and the promoter. We also evaluated copy number variations (by CGH array) and transcriptional changes by RNA Sanger sequencing, whenever possible. This combined strategy revealed 11 likely pathogenic variants in 8 of 10 patients. The molecular diagnosis was fully achieved in 3 of 8 patients, while only one heterozygous mutation was observed in 5 subjects. This approach revealed to be a fast and cost-effective way to analyse the large NEB gene in a small group of patients and might be promising for the detection of pathological variants of other genes featuring large coding regions and lacking mutational hotspots.",
keywords = "NEB mutations, Nemaline myopathy, Next-generation sequencing",
author = "Daniela Piga and Francesca Magri and Dario Ronchi and Stefania Corti and Denise Cassandrini and Eugenio Mercuri and Giorgio Tasca and Enrico Bertini and Fabiana Fattori and Antonio Toscano and Sonia Messina and Isabella Moroni and Marina Mora and Maurizio Moggio and Irene Colombo and Teresa Giugliano and Marika Pane and Chiara Fiorillo and Adele D’Amico and Claudio Bruno and Vincenzo Nigro and Nereo Bresolin and Comi, {Giacomo Pietro}",
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T1 - New Mutations in NEB Gene Discovered by Targeted Next-Generation Sequencing in Nemaline Myopathy Italian Patients

AU - Piga, Daniela

AU - Magri, Francesca

AU - Ronchi, Dario

AU - Corti, Stefania

AU - Cassandrini, Denise

AU - Mercuri, Eugenio

AU - Tasca, Giorgio

AU - Bertini, Enrico

AU - Fattori, Fabiana

AU - Toscano, Antonio

AU - Messina, Sonia

AU - Moroni, Isabella

AU - Mora, Marina

AU - Moggio, Maurizio

AU - Colombo, Irene

AU - Giugliano, Teresa

AU - Pane, Marika

AU - Fiorillo, Chiara

AU - D’Amico, Adele

AU - Bruno, Claudio

AU - Nigro, Vincenzo

AU - Bresolin, Nereo

AU - Comi, Giacomo Pietro

PY - 2016

Y1 - 2016

N2 - Nemaline myopathy represents a group of clinically and genetically heterogeneous neuromuscular disorders. Different clinical-genetic entities have been characterized in the last few years, with implications for diagnostics and genetic counseling. Fifty percent of nemaline myopathy forms are due to NEB mutations, but genetic analysis of this large and complex gene by Sanger sequencing is time consuming and expensive. We selected 10 Italian patients with clinical and biopsy features suggestive for nemaline myopathy and negative for ACTA1, TPM2 and TPM3 mutations. We applied a targeted next-generation sequencing strategy designed to analyse NEB coding regions, the relative full introns and the promoter. We also evaluated copy number variations (by CGH array) and transcriptional changes by RNA Sanger sequencing, whenever possible. This combined strategy revealed 11 likely pathogenic variants in 8 of 10 patients. The molecular diagnosis was fully achieved in 3 of 8 patients, while only one heterozygous mutation was observed in 5 subjects. This approach revealed to be a fast and cost-effective way to analyse the large NEB gene in a small group of patients and might be promising for the detection of pathological variants of other genes featuring large coding regions and lacking mutational hotspots.

AB - Nemaline myopathy represents a group of clinically and genetically heterogeneous neuromuscular disorders. Different clinical-genetic entities have been characterized in the last few years, with implications for diagnostics and genetic counseling. Fifty percent of nemaline myopathy forms are due to NEB mutations, but genetic analysis of this large and complex gene by Sanger sequencing is time consuming and expensive. We selected 10 Italian patients with clinical and biopsy features suggestive for nemaline myopathy and negative for ACTA1, TPM2 and TPM3 mutations. We applied a targeted next-generation sequencing strategy designed to analyse NEB coding regions, the relative full introns and the promoter. We also evaluated copy number variations (by CGH array) and transcriptional changes by RNA Sanger sequencing, whenever possible. This combined strategy revealed 11 likely pathogenic variants in 8 of 10 patients. The molecular diagnosis was fully achieved in 3 of 8 patients, while only one heterozygous mutation was observed in 5 subjects. This approach revealed to be a fast and cost-effective way to analyse the large NEB gene in a small group of patients and might be promising for the detection of pathological variants of other genes featuring large coding regions and lacking mutational hotspots.

KW - NEB mutations

KW - Nemaline myopathy

KW - Next-generation sequencing

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