先天性心脏病(CHD)是一种最为常见的出生缺陷，遗传因素在CHD的发生、发展中具有重要作用，但目前已知的遗传变异只能解释一小部分CHD病例，许多新的CHD相关基因亟待发现。我们在前期研究中，对150例复杂CHD的代表类型心脏圆锥动脉干畸形（CTD）患者样本通过全基因组高分辨芯片分析，发现18个新生拷贝数变异（CNV），并从中遴选出59个新的CHD候选相关基因。本项目拟以这59个候选基因为基础，设计靶向外显子组捕获系统，通过下一代测序技术对500例CTD患者和500例正常对照样本进行CHD新候选相关基因外显子组高通量测序，并利用生物信息学软件进行分析，经Sanger测序法验证，遴选新的CHD相关基因；进一步建立相应的斑马鱼模型，对新鉴定的CHD相关基因进行初步功能研究。本项目的研究，将有利于阐明CHD的遗传发病机制，发现新的CHD遗传生物标志物，为CHD的早期分子诊断和干预治疗提供理论依据。

Congenital heart disease (CHD) is the most common birth defects. Genetic factors play important roles in pathogenesis and development of CHD. However, disease gene mutations identified over the past few decades only explain a small fraction of all CHD cases，many more CHD associated genes are yet to be discovered. In our previous study, we found 18 de novo copy number variations (CNVs) from 150 well clinically characterized patients with a common type of CHD, conotruncal defects (CTD), by using genome-wide high resolution microarray analysis.Fifty-nine CHD candidate genes were identified from the genes within the CNVs.In this study,we plan to develop a next generation sequencing (NGS) based mutation screening system to interrogate sequence variants in 59 CHD candidate genes. We intend to capture and sequence all the coding and exon-intron boundary sequences of 59 CHD candidate genes in samples from 500 clinically evaluated cases with CTD as well as 500 normal controls with Agilent SureSelect Target enrichment system and Miseq sequencing platform. Sequencing data will be assembled and aligned to the reference human genome (hg19). Variants calling, annotation, and evaluation on functional impacts of variants will be performed with a variety of bioinformatic tools. Sanger sequencing will be used to validate the resultant variants.The primary functions of the identified novel CHD gene(s) will be studied by using zebrafish model. The findings of this project will lead to the discovery of new CHD genetic biomarker(s) and help elucidate novel mechanism of CHD pathogenesis as well as facilitate early molecular diagnosis and even prevention of CHD.