Identification of multiple ACVRL1 mutations in patients with pulmonary arterial hypertension by targeted exome capture
Identification of multiple ACVRL1 mutations in patients with pulmonary arterial hypertension by targeted exome capture
复制标题
通过靶向外显子组捕获鉴定肺动脉高压患者的多个 AVRL1 突变。
DOI：
10.1042/cs20160247
复制
发表时间：
2016-09-01
期刊：
影响因子：
6
通讯作者：
Du, Jie
中科院分区：
文献类型：
--
作者：
Piao, Chunmei;Zhu, Yan;Du, Jie
肺动脉高压（PAH）的特点是肺动脉血管系统压力持续升高，可归因于多种原因。此前已有十多个基因被认为与多环芳烃有关。为了检测这些基因在PAH患者中的潜在突变，我们开发了一个包含22个PAH相关基因的靶向外显子组试剂盒，用于80名不相关的PAH患者的遗传筛选。结果，我们在BMPR2基因中发现了16种不同的突变，在激活素受体样激酶1 （ACVRL1）基因ACVRL1中发现了4种不同的突变。然而，在其余20个基因中未发现有害突变。在本研究中，我们提供了四个家系ACVRL1突变的详细特征，包括两个新的错义突变（C . 676g >A, p.V226M; C . 955g >C, p.G319R）和两个复发突变（C . 1231c >T, p.R411W; C . 1450c >T, p.R484W）。此外，我们发现在四个ACVRL1突变体转染的NIH-3T3细胞中，Smad1/5磷酸化水平和荧光素酶报告活性均显著降低。因此，我们的研究结果表明，本研究中发现的ACVRL1错义突变显著影响骨形态发生蛋白9 （BMP-9）通路，暗示PAH发病机制。详细的基因型-表型相关分析显示，一些患者出现遗传性出血性毛细血管扩张症（HHT）的初始症状，提示测序分子标记对于PAH和潜在HHT风险个体的早期识别和干预的重要性。我们开发了一个定制的外显子组测序系统来鉴定这些pah相关基因的突变，并在四个无亲缘关系的中国家庭中发现了两个新的错义突变和两个复发性突变。我们还通过功能研究确定了次胚等位基因。
Pulmonary artery hypertension (PAH) is characterized as sustained elevation of pressure in the pulmonary vascular system that is attributable to a variety of causes. More than a dozen genes have previously been proposed as being associated with PAH. To examine potential mutations of these genes in patients with PAH, we developed a targeted exome kit containing 22 PAH-associated genes for genetic screens of 80 unrelated patients with PAH. As a result, we identified 16 different mutations in the BMPR2 gene and four different mutations in ACVRL1, the gene for activin receptor-like kinase-1 (ACVRL1). However, no deleterious mutations were found in the remaining 20 genes. In the present study, we provided detailed characterization of the ACVRL1 mutations in four pedigrees, including two novel missense mutations (c.676G>A, p.V226M; c.955G>C, p.G319R) and two recurrent mutations (c.1231C>T, p.R411W; c.1450C>T, p.R484W). Furthermore, we showed that markedly reduced Smad1/5 phosphorylation levels and reduced activities of luciferase reporters in each of the four ACVRL1 mutant-transfected NIH-3T3 cells. Therefore, our findings demonstrated that missense mutations of ACVRL1 identified in the present study significantly affected the bone morphogenetic protein 9 (BMP-9) pathway, implicating PAH pathogenesis. Detailed genotype-phenotype correlation analysis revealed initial symptoms of hereditary haemorrhagic telangiectasia (HHT) in some of the patients, suggesting the importance of sequencing molecular markers for early identification and intervention of individuals at risk for PAH and potential HHT. We developed a customized exome sequencing system to identify mutations in these PAH-associated genes, and found two novel missense mutations and two recurrent mutations in the ACVRL1 gene in four unrelated Chinese families; we also determined hypomorphic alleles using functional studies.