西藏地区先天性心脏病分子流行病学研究

Congenital heart disease is one of the most common birth defects throughout the world. Generally, most congenital heart diseases are thought to have a multifactorial origin, with interaction of genetic and environmental exposures. However, differences in genetic heterogeneity and genetic polymorphism among different ethnic groups are significant. Studies have demonstrated that the prevalence of congenital heart disease in Tibetan areas was significantly higher than that in plain areas. In the present study, Affymetrix Cytoscan HD Array Technology was used to explore the relationship between genome-wide single nucleotide polymorphisms and copy number variation and susceptibility to congenital heart diseases in parents and offspring in different groups. SequenomMassARRAY SNP and Real Time PCR were used to validate the SNP and CNV sites in a large sample population. Moreover, the functions of SNP were predicted by bioinformatics tools. The aim of this study is to provide the evidence for further prospective study and personalized prevention of congenital heart diseases.

先天性心脏病是人类最常见和最严重的新生儿出生缺陷性疾病之一，目前多数研究认为先天性心脏病是由遗传因素和环境因素共同作用所致的复杂性疾病。但不同的种族人群间遗传异质性和遗传多态性的差别较大，调查显示西藏地区藏族人群先天性心脏病发病率显著高于平原地区。本课题组拟采用病例对照研究方法，通过Affymetrix Cytoscan HD Array基因芯片技术探讨西藏地区藏/汉族先天性心脏病患者及健康对照及父母的全基因组单核苷酸多态性(SNP)和拷贝数变异(CNV)与先天性心脏病易感性的关系；并采用SequenomMassARRAY SNP分型技术和Real Time PCR技术在大样本人群中验证筛查到的SNP和CNV位点，并通过生物信息学手段对SNP进行功能学预测，同时探讨西藏环境因素与个体的遗传易感性间的交互作用，为进一步阐明先天性心脏病致病机制及开展先天性心脏病的个性化预防提供依据。

先天性心脏病是最常见的新生儿出生缺陷性疾病之一，目前多数研究认为先天性心脏病是由遗传因素和环境因素共同作用所致的复杂性疾病。本课题组应用SureSelectXT Human全外显子捕获试剂盒进行全外显子捕获，采用Illumina HiSeq平台进行2×150 bp双末端测序模式高通量测序探讨西藏地区藏族17例房间隔缺损（ASD）患者，18例卵圆孔未闭患者和8例室间隔缺损（VSD）患儿及其父母的差异基因单核苷酸多态性(SNP)易感性的关系；采用GATK HaplotypeCaller软件推荐的筛查方案检测每个样本的SNV/InDel，并通过生物信息学手段对SNP位点进行功能学预测。最终筛选出6个与VSD发生相关的遗传性致病基因突变，分别为ACVRL1（c.652C>T）、ATP13A2（c.1363C>T）、ERAP1（c.481G>A）、MRI1（c.629G>A）、TRAP1（c.224G>A）和FBN2（c.2260G>A）；7个与ASD相关的致病突变，分别为CEMIP2（c.2732G>A：p.R911H）、CRYAB（c.269G>A）、ERAP1（c.481G>A）、HLCS（c.1148A>G）、ITK（c.1741C>T）、NTN3（c.1243C>T：p.R415C）、SLC45A2（c.626C>T）；7个与卵圆孔未闭发生相关的致病突变，分别为ATP13A5（c.1967G>A）、GABRP（c.421C>T：p.R141C）、MMP24（c.389C>T）、SLC45A2（c.626C>T）、STIM1（c.2246G>A）、THBS4（c.2195C>T）、TRPM2（c.3433G>A）。我们首次应用全外显子组测序技术直接筛查了西藏高原地区先天性心脏病的遗传变异，为进一步阐明先天性心脏病致病机制及开展先天性心脏病的个性化预防提供依据。