Next-generation sequencing application to investigate skeletal muscle channelopathies in a large cohort of Italian patients.

• Skeletal muscle channelopathies are rare genetic diseases. • NGS screening is a powerful tool in the diagnostic workflow for skeletal muscle channelopathies. • The CLCN1 and SCN4A are mainly genes associated with non-dystrophic myotonia. • Collaboration between clinicians and molecular geneticists to make a diagnosis of SMC. Non-dystrophic myotonias and periodic paralyses are a heterogeneous group of disabling diseases classified as skeletal muscle channelopathies. Their genetic characterization is essential for prognostic and therapeutic purposes; however, several genes are involved. Sanger-based sequencing of a single gene is time-consuming, often expensive; thus, we designed a next-generation sequencing panel of 56 putative candidate genes for skeletal muscle channelopathies, codifying for proteins involved in excitability, excitation-contraction coupling, and metabolism of muscle fibres. We analyzed a large cohort of 109 Italian patients with a suspect of NDM or PP by next-generation sequencing. We identified 24 patients mutated in CLCN1 gene, 15 in SCN4A , 3 in both CLCN1 and SCN4A , 1 in ATP2A1 , 1 in KCNA1 and 1 in CASQ1. Eight were novel mutations: p.G395Cfs*32, p.L843P, p.V829M, p.E258E and c.1471+4delTCAAGAC in CLCN1 , p.K1302R in SCN4A , p.L208P in ATP2A1 and c.280–1G>C in CASQ1 genes. This study demonstrated the utility of targeted next generation sequencing approach in molecular diagnosis of skeletal muscle channelopathies and the importance of the collaboration between clinicians and molecular geneticists and additional methods for unclear variants to make a conclusive diagnosis. [ABSTRACT FROM AUTHOR]