ABSTRACT Non-dystrophic myotonias (NDM) and periodic paralyses (PP) are a heterogeneous group of disabling diseases classified as skeletal muscle channelopathies (SMC). 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 SMC, 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.

中文翻译：

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用于研究大量意大利患者的骨骼肌通道病的新一代测序应用程序

摘要 非营养不良性肌强直 (NDM) 和周期性麻痹 (PP) 是一组异质性致残疾病，归类为骨骼肌通道病 (SMC)。它们的遗传特征对于预后和治疗目的至关重要；然而，涉及几个基因。基于 Sanger 的单个基因测序既耗时又昂贵；因此，我们为 SMC 设计了一个包含 56 个推定候选基因的下一代测序面板，编码了参与兴奋性、兴奋-收缩耦合和肌肉纤维代谢的蛋白质。我们通过下一代测序分析了 109 名疑似 NDM 或 PP 的意大利患者。我们确定了 CLCN1 基因突变的 24 名患者，SCN4A 突变的患者 15 名，CLCN1 和 SCN4A 突变患者 3 名，ATP2A1 突变患者 1 名，KCNA1 突变患者 1 名，CASQ1 突变患者 1 名。八个是新的突变：p.G395Cfs*32，p。CLCN1 中的 L843P、p.V829M、p.E258E 和 c.1471+4delTCAAGAC，SCN4A 中的 p.K1302R，ATP2A1 中的 p.L208P 和 CASQ1 基因中的 c.280-1G>C。这项研究证明了靶向下一代测序方法在骨骼肌通道病分子诊断中的效用，以及临床医生和分子遗传学家之间合作的重要性，以及对不清楚变异的其他方法进行结论性诊断的重要性。