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.

New Mutations in NEB Gene Discovered by Targeted Next-Generation Sequencing in Nemaline Myopathy Italian Patients / D. Piga, F. Magri, D. Ronchi, S. Corti, D. Cassandrini, E. Mercuri, G. Tasca, E. Bertini, F. Fattori, A. Toscano, S. Messina, I. Moroni, M. Mora, M. Moggio, I. Colombo, T. Giugliano, M. Pane, C. Fiorillo, A. D’Amico, C. Bruno, V. Nigro, N. Bresolin, G.P. Comi. - In: JOURNAL OF MOLECULAR NEUROSCIENCE. - ISSN 0895-8696. - 59:3(2016), pp. 351-359. [10.1007/s12031-016-0739-2]

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

F. Magri
Secondo
;
D. Ronchi;S. Corti;I. Colombo;N. Bresolin
Penultimo
;
G.P. Comi
2016

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.
NEB mutations; Nemaline myopathy; Next-generation sequencing
Settore MED/26 - Neurologia
Settore MED/03 - Genetica Medica
Settore BIO/18 - Genetica
22-apr-2016
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/398197
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