Next generation sequencing on patients with LGMD and nonspecific myopathies: Findings associated with ANO5 mutations.
Next generation sequencing on patients with LGMD and nonspecific myopathies: Findings associated with ANO5 mutations.
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DOI：
10.1016/j.nmd.2015.03.011
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发表时间：
2015-07
期刊：
影响因子：
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通讯作者：
Nigro V
中科院分区：
文献类型：
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作者：
Savarese M;Di Fruscio G;Tasca G;Ruggiero L;Janssens S;De Bleecker J;Delpech M;Musumeci O;Toscano A;Angelini C;Sacconi S;Santoro L;Ricci E;Claes K;Politano L;Nigro V
我们对ANO 5基因进行了最大规模的筛选。我们确定了33例患者（4%）的致病性变化，在两个等位基因和23个杂合子（3%）。ANO 5携带者的鉴定并不被认为是一个罕见的发现。anoctaminopathies具有极高的遗传和表型异质性。NGS为基础的策略是完美的解剖NMD的临床变异性。我们研究了786例未确诊的LGMD或非特异性肌病特征患者，以使用下一代测序（NGS）方法研究ANO 5突变在肢带型肌营养不良症（LGMD）和非特异性肌病中的作用。在160例LGMD患者中，我们首先对热点外显子5和20进行测序，然后对编码区的剩余部分进行测序。另外626例患者，采用更广泛的纳入标准招募，直接通过靶向NGS进行分析。通过结合NGS和桑格测序，我们确定了33/786（4%）携带两个等位基因和23个ANO 5杂合子（3%）推定的致病性变化的患者。表型谱比预期的更广，从高CK血症到肌病，缺乏基因型/表型相关性。特别是，这是目前对ANO 5基因最大规模的筛选。大量的破坏性突变的杂合子提示anoctaminopathies应该是频繁的，往往是非渗透性的。我们建议通过靶向NGS进行多重基因检测，作为分析非特异性肌病表现患者的第一步。这代表了一种简单的方法来克服ANO 5患者临床异质性的困难，并同时测试与神经肌肉疾病有关的许多其他基因。
We have carried out the largest screening of the ANO5 gene. We identified 33 patients (4%) with pathogenic changes in both alleles and 23 heterozygotes (3%). The identification of a ANO5 carrier is not to be considered an uncommon finding. The anoctaminopathies have an extremely high genetic and phenotypic heterogeneity. NGS-based strategies are perfect to dissect the clinical variability in NMDs. 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.