RNA sequencing solved the most common but unrecognized NEB pathogenic variant in Japanese nemaline myopathy
RNA sequencing solved the most common but unrecognized NEB pathogenic variant in Japanese nemaline myopathy
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DOI：
10.1038/s41436-018-0360-6
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发表时间：
2019-07-01
影响因子：
8.8
通讯作者：
Matsumoto, Naomichi
中科院分区：
文献类型：
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作者：
Hamanaka, Kohei;Miyatake, Satoko;Matsumoto, Naomichi
目的：外显子组测序对孟德尔病的诊断率一般为20-40%。发生率低的部分原因是ES错过了导致异常剪接的深层内含子或同义变体。本研究旨在应用RNA序列分析技术(RNA-seq)有效地检测异常剪接及其相关变异。方法：对6例ES未解决的线状肌病(NM)患者活体肌肉组织中的异常剪接进行RNA-seq分析。用Sanger测序分析与检测到的异常剪接事件相关的变体。结果：在1例新发现的深内含子NEB致病变异体c.1569+339A>；G和3例新的同义NEB致病变异体c.24684G>；C(p.Ser8228Ser)。在正常日本人群中，C.24684G>；C变异是所有NEB致病变异中最常见的，频率为1/178(3552人中有20个等位基因)，但此前未被识别。对变异体的扩大筛查在另外四个先前未解决的线虫肌病病例中发现了它。结论：这些结果表明，RNA-SEQ可能能够解决很大一部分先前未被诊断的肌肉疾病。
Purpose: The diagnostic rate for Mendelian diseases by exome sequencing (ES) is typically 20-40%. The low rate is partly because ES misses deep-intronic or synonymous variants leading to aberrant splicing. In this study, we aimed to apply RNA sequencing (RNA-seq) to efficiently detect the aberrant splicings and their related variants.Methods: Aberrant splicing in biopsied muscles from six nemaline myopathy (NM) cases unresolved by ES were analyzed with RNA-seq. Variants related to detected aberrant splicing events were analyzed with Sanger sequencing. Detected variants were screened in NM patients unresolved by ES.Results: We identified a novel deep-intronic NEB pathogenic variant, c.1569+339A>G in one case, and another novel synonymous NEB pathogenic variant, c.24684G>C (p.Ser8228Ser) in three cases. The c.24684G>C variant was observed to be the most frequent among all NEB pathogenic variants in normal Japanese populations with a frequency of 1 in 178 (20 alleles in 3552 individuals), but was previously unrecognized. Expanded screening of the variant identified it in a further four previously unsolved nemaline myopathy cases.Conclusion: These results indicated that RNA-seq may be able to solve a large proportion of previously undiagnosed muscle diseases.