Somatic mutations in cerebral cortical malformations.
Somatic mutations in cerebral cortical malformations.
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DOI：
10.1056/nejmoa1314432
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发表时间：
2014-08-21
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影响因子：
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通讯作者：
Walsh CA
中科院分区：
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作者：
Jamuar SS;Lam AT;Kircher M;D'Gama AM;Wang J;Barry BJ;Zhang X;Hill RS;Partlow JN;Rozzo A;Servattalab S;Mehta BK;Topcu M;Amrom D;Andermann E;Dan B;Parrini E;Guerrini R;Scheffer IE;Berkovic SF;Leventer RJ;Shen Y;Wu BL;Barkovich AJ;Sahin M;Chang BS;Bamshad M;Nickerson DA;Shendure J;Poduri A;Yu TW;Walsh CA
尽管越来越多的人认识到体细胞突变在遗传疾病中的作用，但体细胞突变在神经发育疾病中的流行程度以及检测体细胞嵌合体的最佳技术尚未得到系统评估。使用与脑畸形相关的已知和候选基因的定制小组，我们对来自158名脑畸形患者的白细胞来源的≥样本进行了有针对性的高覆盖测序(深度，Depth，DNA200×)，包括双皮质综合征(皮质下带状异位，30人)，多小脑回伴巨脑畸形(20人)，脑室周围结节异位(61人)，和厚脑回(47人)。我们使用Sanger测序验证了候选突变，对于存在不同阅读深度的变异体，亚克隆后进行菌落测序。经过验证，在27人中发现了因果突变(17%；每种表型的突变范围为10%至30%)。在27名患者中有8名(30%)是体细胞突变，主要发生在双皮质综合征患者(我们在这些患者中发现了DCX和LIS1突变)、脑室周围结节异位症(FLNA)和粗脑回患者(TUBB2B)。在我们检测到的体细胞突变中，有5个(63%)用传统的Sanger测序无法检测到，但通过亚克隆和随后的亚克隆DNA测序得到了验证。我们在粗脑回患者中发现了候选基因DYNC1H1、KIF5C和其他动蛋白基因的潜在因果突变。靶向测序被发现对检测脑畸形患者的体细胞突变是有用的。高覆盖率测序小组在评估神经精神疾病的体细胞突变方面提供了对全外显子组和全基因组测序的重要补充。(由国家神经疾病和中风研究所等资助。)
Although there is increasing recognition of the role of somatic mutations in genetic disorders, the prevalence of somatic mutations in neurodevelopmental disease and the optimal techniques to detect somatic mosaicism have not been systematically evaluated. Using a customized panel of known and candidate genes associated with brain malformations, we applied targeted high-coverage sequencing (depth, ≥200×) to leukocyte-derived DNA samples from 158 persons with brain malformations, including the double-cortex syndrome (subcortical band heterotopia, 30 persons), polymicrogyria with megalencephaly (20), periventricular nodular heterotopia (61), and pachygyria (47). We validated candidate mutations with the use of Sanger sequencing and, for variants present at unequal read depths, subcloning followed by colony sequencing. Validated, causal mutations were found in 27 persons (17%; range, 10 to 30% for each phenotype). Mutations were somatic in 8 of the 27 (30%), predominantly in persons with the double-cortex syndrome (in whom we found mutations in DCX and LIS1), persons with periventricular nodular heterotopia (FLNA), and persons with pachygyria (TUBB2B). Of the somatic mutations we detected, 5 (63%) were undetectable with the use of traditional Sanger sequencing but were validated through subcloning and subsequent sequencing of the subcloned DNA. We found potentially causal mutations in the candidate genes DYNC1H1, KIF5C, and other kinesin genes in persons with pachygyria. Targeted sequencing was found to be useful for detecting somatic mutations in patients with brain malformations. High-coverage sequencing panels provide an important complement to whole-exome and whole-genome sequencing in the evaluation of somatic mutations in neuropsychiatric disease. (Funded by the National Institute of Neurological Disorders and Stroke and others.)