Contribution of intragenic deletions to mutation spectrum in Chinese patients with Wilson's disease and possible mechanism underlying ATP7B gross deletions
Contribution of intragenic deletions to mutation spectrum in Chinese patients with Wilson's disease and possible mechanism underlying ATP7B gross deletions
复制标题
基因内缺失对中国威尔逊病患者突变谱的贡献及ATP7B总缺失的可能机制
DOI：
10.1016/j.parkreldis.2019.01.001
复制
发表时间：
2019-05-01
影响因子：
4.1
通讯作者：
Wu, Zhi-Ying
中科院分区：
文献类型：
--
作者：
Chen, Yu-Chao;Yu, Hao;Wu, Zhi-Ying
简介：威尔逊病（WD）是一种常染色体隐性遗传的铜代谢疾病，由于ATP 7B致病突变。如果早期诊断和有效治疗，疾病症状是可以预防的。直接测序是常规用于确认WD的诊断，但不能确定总rearrangement.Methods：桑格序列的ATP 7B进行了142个新招募的WD指数患者。根据美国医学遗传学和基因组学学院（ACMG）标准和指南对鉴定的变体的临床效果进行分类。多重连接依赖性探针扩增（MLPA）在168例临床WD无法解释的桑格测序，从我们的总病例系列的774例WD患者中选择。在确定总重排内ATP 7B，断裂点进行了测定，通过长距离PCR和直接sequencing.Results：在142 WD患者，我们确定了71个序列改变ATP 7B，其中15个是新的，其中14个被归类为“致病性”或“可能致病性”，包括2个影响剪接位点的内含子变异。在168例WD患者中的6例中，MLPA确定了4个杂合性总ATP 7B缺失。结论：WD的发生与基因内缺失有关，非同源末端连接可能是WD的发病机制之一，因此，WD的综合基因检测应包括基因内缺失的检测。
Introduction: Wilson's disease (WD) is an autosomal recessive disorder of copper metabolism due to ATP7B pathogenic mutations. Disease manifestations can be prevented if early diagnosis and effective treatment are given. Direct sequencing is routinely used to confirm WD diagnosis, but cannot identify gross rearrangements.Methods: Sanger sequencing of ATP7B was performed in 142 newly recruited WD index patients. The clinical effects of identified variants were classified according to American College of Medical Genetics and Genomics (ACMG) standards and guidelines. Multiplex ligation-dependent probe amplification (MLPA) was performed in 168 WD cases with clinical WD unexplained by Sanger sequencing, selected from our total case series of 774 WD patients. After identifying gross rearrangements within ATP7B, the breakpoints were determined by long-range PCR and direct sequencing.Results: In the 142 WD patients, we identified 71 sequence alterations in ATP7B, of which 15 were novel; 14 of these were classified as 'pathogenic' or 'likely pathogenic', including 2 intronic variants affecting splice sites. In 6 of 168 WD patients, MLPA identified four heterozygous gross ATP7B deletions. One was a whole gene deletion, and three were intragenic deletions which were mapped to breakpoint locations, revealing non-homologous end joining.Conclusion: Intragenic deletions are responsible for WD and non-homologous end joining could be the pathogenesis, therefore the detection of intragenic deletions should be included in comprehensive genetic testing for WD.