ARRA - NHLBI Lung Cohorts Sequencing Project

描述（由申请人提供）：项目摘要：本申请响应NHLBI参与研究和研究基础设施“重大机遇”（RC2） （RFA-OD-09-004）的大规模DNA测序和表型良好的NHLBI队列的分子分析。该提案的主要目标是应用下一代重测序来识别影响儿科和成人肺部疾病的致病变异，这是美国第三大死亡原因。我们将应用的技术涉及人类基因组（“外显子组”）中所有蛋白质编码序列的大规模平行重测序。在最初的发现阶段，我们将与选定的测序中心合作，从主要临床表型分布的每个尾部（总共1400个外显子组）中选择50-300个个体，生成罕见和常见变异的目录，评估七个人群队列中的每个人群，其中一套全面的临床特征已经得到很好的表征。我们的表型包括：囊性纤维化患者肺部疾病的严重程度、囊性纤维化患者铜绿假单胞菌（Pa）的获得时间、哮喘患者肺部疾病的严重程度、慢性阻塞性肺疾病患者肺功能下降的比率、肺动脉高压的严重程度和急性肺损伤的严重程度。每条尾巴的变异目录将通过新的生物信息学方法进行挖掘，以确定高优先级的疾病修饰基因和/或因果变异。这种全基因组重测序方法是现代遗传学中最重要的挑战和机遇之一。外显子组测序可以揭示目前未被全基因组关联研究分型或检测到的低频等位基因。在第二阶段，我们建议使用常规方法在每个队列中的所有个体中评估每个高优先级候选基因或变体。发现影响这些重叠肺部表型的遗传变异将大大扩展我们对肺部疾病生物学的理解，并将促进对影响我国最年幼和最年长的一组疾病的准确诊断和改进管理。我们的发现很可能为美国和全球预防肺部疾病的新疗法提供见解。

DESCRIPTION (provided by applicant): Project Summary This application responds to NHLBI Participation in Research and Research Infrastructure "Grand Opportunities" (RC2) (RFA-OD-09-004) for Large-scale DNA Sequencing and Molecular Profiling of Well-Phenotyped NHLBI Cohorts. The major goal of this proposal is to apply next-generation resequencing to identify disease-causing variants influencing a key set of pediatric and adult lung diseases, the third leading cause of death in the United States. The technology we will apply involves massively parallel resequencing of all protein coding sequences in the human genome (the "exome"). In the initial discovery stage we will work with a selected sequencing center(s) to generate a catalogue of rare and common variants from 50-300 individuals selected from each of the tails of the distribution (1400 exomes total) of the major clinical phenotype assessed for each of seven population cohorts in which a comprehensive set of clinical traits have been well-characterized. Our phenotypes include: severity of lung disease in cystic fibrosis, time to acquisition of Pseudomonas aeruginosa (Pa) in cystic fibrosis, severity of lung disease in asthma, rate of decline of lung function in chronic obstructive pulmonary disease, severity of pulmonary hypertension and severity of acute lung injury. The variant catalogues from each tail will be mined by novel bioinformatics approaches to identify high-priority disease-modifying genes and/or causal variants. This genome-wide resequencing approach presents one of the most important challenges and opportunities in modern genetics. Exome sequencing can uncover low frequency alleles not currently typed or detectable by genome-wide association studies. In a second stage, we propose each high priority candidate gene or variant will be evaluated in all individuals in each cohort using conventional methods. Discovery of the genetic variants influencing these overlapping pulmonary phenotypes will substantially expand our understanding of biology of lung diseases, and will facilitate accurate diagnosis and improved management of a group of diseases that impact the very youngest and oldest in our country. Our findings may well provide insights for novel therapeutics to prevent lung disease in the U.S. and globally. PUBLIC HEALTH RELEVANCE: Project Narrative The major goal of this project is to identify the gene variations that influence the severity of lung diseases such as asthma, chronic obstructive pulmonary disease, acute lung injury, pulmonary arterial hypertension, and cystic fibrosis. This goal will be accomplished by using a novel genome-wide approach that compares the DNA sequence of every human gene among 1400 individuals of diverse ethnic ancestry. Finding these genes will improve our understanding of the biology of these acute and chronic lung diseases, improve their management, and provide information for the development of novel therapeutics.