Exome sequencing and functional studies in familial CHD

描述(由申请人提供)： 大规模并行测序技术的发展可能掌握着个性化医学的关键，因为它允许识别使人易患疾病的功能性遗传变异。为了使这成为现实，必须确定致病基因。这项建议集中在非综合征性先天性心脏病(CHDS)。先天性心脏病是最常见的出生缺陷类型，不仅影响儿童，而且影响越来越多的成年幸存者。流行病学研究已经证实了遗传因素，但几乎没有发现致病基因。在这一认识鸿沟被克服之前，潜在的分子原因仍然是隐藏的，并阻碍新疗法的开发和潜在的长期并发症的预测。我们的长期目标是确定潜在的遗传原因，并阐明导致CHD的分子机制。这项应用的总体目标是发现导致冠心病的遗传变异。这一中心假设是，可以通过全基因组测序的方法，利用表现出孟德尔遗传模式的家族来识别导致CHD的基因。这项拟议研究的基本原理是，发现先天性心脏病的遗传原因有可能提供更好的风险咨询，并导致导致死亡的畸形的新疗法。 从婴儿期到成年期。在支持这一前提的最新文献的指导下，并拥有一个独特的家系队列，中心假设将通过追求两个具体目标来检验：1)扩大我们目前的多基因家系和散发性CHD病例队列；2)通过外显子组和全基因组测序的传递不平衡测试，在表现出孟德尔分离的CHD家系和三联体中识别致病基因。将专门开发方法来测试TRIO数据中罕见的变异关联，这些关联聚合在一个基因区域内的变异上，并根据频率和功能对这些变异进行加权。这些方法还将允许对有家族病史的无关个人进行联合分析。 CHD和TRIO数据。推测的因果变异的功能效应将通过体外和体内的非洲爪哇试验进行研究。然后，将对更大的冠心病队列进行筛查，以寻找已识别基因的突变。我们的方法是创新的，不仅因为它采用了新技术，而且通过使用家族来解决复杂疾病的遗传基础，增加了孟德尔遗传学的力量。这项应用侧重于NHLBI战略计划的一部分，目标和挑战，以确定人类群体中与特定疾病相关的关键遗传变异。这一提议的成功将在人类疾病基因识别方法上创造一种范式转变。这项拟议的研究具有重要意义，因为它有望通过识别CHD的病因基因，垂直推进人类遗传学、发育生物学和心血管医学领域的发展。在当前的胎儿诊断和干预时代，这些知识将被用于改进预防措施、宫内诊断、遗传咨询和治疗。 公共卫生相关性： 项目叙事。拟议的研究与公共卫生有关，因为它解决了婴儿期发病率和死亡率的一个重要原因。这项有计划的研究有可能增加对疾病发病机制的基本了解，并及早识别高危个体以进行可能的干预。因此，拟议的研究与NHLBI战略计划的目标1有关，目的是通过确定导致特定疾病易感性的关键基因变异，增加对健康和疾病的分子和生理基础的了解，并与NICHD通过出生缺陷的基因研究了解疾病易感性的遗传学和正常和异常发育的科学目标相关。

DESCRIPTION (provided by applicant): The development of massively parallel sequencing technologies may hold the key to personalized medicine as it allows for the identification of functional genetic variants that rende a person susceptible to disease. For this to become reality, disease-causing genes must be identified. This proposal concentrates on non-syndromic congenital heart defects (CHDs). Representing the most common type of birth defect, CHDs affect not only children but also a growing population of adult survivors. Epidemiologic studies have demonstrated genetic contributors, but few etiologic genes have been identified. Until this knowledge gap is overcome, the underlying molecular cause will remain hidden and hinder the development of new therapies and potentially prediction of long-term complications. Our long-term goal is to identify the underlying genetic causes and elucidate the molecular mechanisms leading to CHDs. The overall objective of this application is to discover the genetic variation that results in CHDs. Th central hypothesis is that CHD-causing genes can be identified by a genome-wide sequencing approach, using families exhibiting Mendelian inheritance patterns. The rationale for the proposed research is that the discovery of genetic causes of CHDs has the potential to provide better risk counseling and result in novel therapies for malformations that contribute to mortality from infancy to adulthood. Guided by recent literature supporting this premise and possession of a unique cohort of families, the central hypothesis will be tested by pursuing two Specific Aims: 1) Expand our current cohort of multiplex families and sporadic cases of CHDs; 2) Identify disease-causing genes in families exhibiting Mendelian segregation for CHDs and in trios via transmission disequilibrium tests by exome and whole genome sequencing. Methods will be developed specifically to test for rare variant associations in trio data, which aggregate over variants within a gene region and weight these variants based upon frequency and functionality. These methods will also allow for the joint analysis of unrelated individuals with a family history of CHD and trio data. The putative causal variants functional effects will be investigated by in vitro and in vivo Xenopus assays. A larger cohort with CHD will then be screened for mutations in identified genes. Our approach is innovative not only because it employs new technologies but also adds the power of Mendelian genetics through the use of families to tackle the genetic basis of a complex disease. This application focuses on part of the NHLBI Strategic Plan goals and challenges to identify key genetic variants in the human population that are associated with specific diseases. Success of this proposal will create a paradigm shift in the approach for human disease gene identification. The proposed research is significant because it is expected to vertically advance the fields of human genetics, developmental biology and cardiovascular medicine by identifying causal genes for CHDs. In the current era of fetal diagnosis and intervention, this knowledge will be used to improve prevention measures, in utero diagnosis, genetic counseling and therapy. PUBLIC HEALTH RELEVANCE: Project Narrative. The proposed study is relevant to public health as it address a significant cause of morbidity and mortality in infancy. This planned research has the potential to increase the fundamental understanding of disease pathogenesis, and identify at risk individuals early for possible interventions. Thus, the proposed research is relevant to Goal 1 of the NHLBI Strategic Plan, to increase understanding of the molecular and physiological basis of health and disease, by determining key genetic variants that account for susceptibility to specific disease, and to the NICHD's scientific goals of understanding genetics of disease susceptibility and normal and abnormal development by genetic studies of birth defects.