Targeted Next-Generation Sequencing for the Molecular Genetic Diagnostics of Cardiomyopathies
Targeted Next-Generation Sequencing for the Molecular Genetic Diagnostics of Cardiomyopathies
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DOI：
10.1161/circgenetics.110.958322
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发表时间：
2011-04-01
影响因子：
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通讯作者：
Rottbauer, Wolfgang
中科院分区：
文献类型：
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作者：
Meder, Benjamin;Haas, Jan;Rottbauer, Wolfgang
今天，已经发现30多种不同基因的突变可导致遗传性心肌病，其中一些与预后非常差有关。然而，由于遗传异质性和目前诊断工具的吞吐量和可扩展性的限制，到目前为止，很难以一种快速、全面和经济有效的方式对心肌病患者进行遗传表征。方法和结果：我们建立了一种基于阵列的亚基因组富集，然后进行下一代测序，以检测肥厚型心肌病（HCM）和扩张型心肌病（DCM）患者的突变。通过这种方法，我们发现，与全基因组的覆盖范围相比，感兴趣的基因组区域可以被平均富集2169倍，从而导致所选疾病基因的高序列覆盖率，并允许我们在单次测序运行中定义心肌病的遗传发病机制。在6例患者中，我们检测到致病突变，2个微缺失和4个点突变。此外，我们发现了几个新的非同义变异，这些变异被预测是有害的，因此可能是DCM或HCM的潜在疾病突变或修饰因子。结论：本文提出的方法首次允许对遗传性DCM或HCM患者进行快速和经济有效的全面遗传筛查。（中国心血管杂志，2011;4:110-122）
Background-Today, mutations in more than 30 different genes have been found to cause inherited cardiomyopathies, some associated with very poor prognosis. However, because of the genetic heterogeneity and limitations in throughput and scalability of current diagnostic tools up until now, it is hardly possible to genetically characterize patients with cardiomyopathy in a fast, comprehensive, and cost-efficient manner.Methods and Results-We established an array-based subgenomic enrichment followed by next-generation sequencing to detect mutations in patients with hypertrophic cardiomyopathy (HCM) and dilated cardiomyopathy (DCM). With this approach, we show that the genomic region of interest can be enriched by a mean factor of 2169 compared with the coverage of the whole genome, resulting in high sequence coverage of selected disease genes and allowing us to define the genetic pathogenesis of cardiomyopathies in a single sequencing run. In 6 patients, we detected disease-causing mutations, 2 microdeletions, and 4 point mutations. Furthermore, we identified several novel nonsynonymous variants, which are predicted to be harmful, and hence, might be potential disease mutations or modifiers for DCM or HCM.Conclusions-The approach presented here allows for the first time a comprehensive genetic screening in patients with hereditary DCM or HCM in a fast and cost-efficient manner. (Circ Cardiovasc Genet. 2011;4:110-122.)