title: Next generation sequencing on patients with LGMD and nonspecific myopathies: Findings associated with ANO5 mutations.
abstract: We studied 786 undiagnosed patients with LGMD or nonspecific myopathic features to investigate the role of ANO5 mutations in limb-girdle muscular dystrophies (LGMDs) and in nonspecific myopathies using the next generation sequencing (NGS) approach. In 160 LGMD patients, we first sequenced hotspot exons 5 and 20 and then sequenced the remaining part of the coding region. Another 626 patients, recruited using broader inclusion criteria, were directly analyzed by targeted NGS. By combining NGS and Sanger sequencing, we identified 33/786 (4%) patients carrying putative pathogenic changes in both alleles and 23 ANO5 heterozygotes (3%). The phenotypic spectrum is broader than expected, from hyperCKemia to myopathies, with lack of genotype/phenotype correlations. In particular, this is currently the largest screening of the ANO5 gene. The large number of heterozygotes for damaging mutations suggests that anoctaminopathies should be frequent and often nonpenetrant. We propose the multiple genetic testing by targeted NGS as a first step to analyze patients with nonspecific myopathic presentations. This represents a straightforward approach to overcome the difficulties of clinical heterogeneity of ANO5 patients, and to test, at the same time, many other genes involved in neuromuscular disorders.
authors.affiliation: Telethon Institute of Genetics and Medicine, Pozzuoli (NA), Italy; Dipartimento di Biochimica, Biofisica e Patologia Generale, Seconda Università di Napoli, Napoli, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Savarese
authors.affiliation: Telethon Institute of Genetics and Medicine, Pozzuoli (NA), Italy; Dipartimento di Biochimica, Biofisica e Patologia Generale, Seconda Università di Napoli, Napoli, Italy.
authors.forename: Giuseppina
authors.initials: G
authors.lastname: Di Fruscio
authors.affiliation: Fondazione Don Gnocchi, Italy.
authors.forename: Giorgio
authors.initials: G
authors.lastname: Tasca
authors.affiliation: Dipartimento di Neuroscienze e Scienze riproduttive ed odontostomatologiche, Università di Napoli "Federico II", Napoli, Italy.
authors.forename: Lucia
authors.initials: L
authors.lastname: Ruggiero
authors.affiliation: Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium.
authors.forename: Sandra
authors.initials: S
authors.lastname: Janssens
authors.affiliation: Department of Neurology, Ghent University Hospital, Ghent, Belgium.
authors.forename: Jan
authors.initials: J
authors.lastname: De Bleecker
authors.affiliation: Biochimie et génétique moléculaire, Centre hospitalier Cochin, Paris, France.
authors.forename: Marc
authors.initials: M
authors.lastname: Delpech
authors.affiliation: Dipartimento di Neuroscienze, Università di Messina, Messina, Italy.
authors.forename: Olimpia
authors.initials: O
authors.lastname: Musumeci
authors.affiliation: Dipartimento di Neuroscienze, Università di Messina, Messina, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Toscano
authors.affiliation: Dipartimento di Neuroscienze, Università di Padova, Padova, Italy.
authors.forename: Corrado
authors.initials: C
authors.lastname: Angelini
authors.affiliation: Centre de Référence Maladies Neuromusculaires - SLA, Hôpital Archet 1, CHU de Nice, Nice, France.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Sacconi
authors.affiliation: Dipartimento di Neuroscienze e Scienze riproduttive ed odontostomatologiche, Università di Napoli "Federico II", Napoli, Italy.
authors.forename: Lucio
authors.initials: L
authors.lastname: Santoro
authors.affiliation: Fondazione Don Gnocchi, Italy.
authors.forename: Enzo
authors.initials: E
authors.lastname: Ricci
authors.affiliation: Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium.
authors.forename: Kathleen
authors.initials: K
authors.lastname: Claes
authors.affiliation: Dipartimento di Medicina Sperimentale, Seconda Università di Napoli, Napoli, Italy.
authors.forename: Luisa
authors.initials: L
authors.lastname: Politano
authors.affiliation: Telethon Institute of Genetics and Medicine, Pozzuoli (NA), Italy; Dipartimento di Biochimica, Biofisica e Patologia Generale, Seconda Università di Napoli, Napoli, Italy. Electronic address: vinnigro@gmail.com.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Nigro
mesh_terms: D000328:Adult; D000368:Aged; D000483:Alleles; D000075368:Anoctamins; D018118:Chloride Channels / Q000235:genetics*; D015331:Cohort Studies; D005091:Exons; D005260:Female; D006579:Heterozygote; D006801:Humans; D008297:Male; D008875:Middle Aged; D009135:Muscular Diseases / Q000235:genetics*; D049288:Muscular Dystrophies, Limb-Girdle / Q000235:genetics*; D009154:Mutation; D010641:Phenotype; D017422:Sequence Analysis, DNA; D055815:Young Adult
chemical_list: C547813:ANO5 protein, human; D000075368:Anoctamins; D018118:Chloride Channels
keywords: Anoctamin; LGMD2L; Limb girdle muscular dystrophy; Muscular dystrophy; NGS screening; Next generation sequencing; Targeted resequencing