Tissue-based validation of COPD Genetic Studies using Lung Health Study Cohort

描述（由申请人提供）： 慢性阻塞性肺疾病（COPD）是北美第四大死亡原因，影响着1000万至1200万美国人。虽然吸烟是COPD发展中最常见的环境风险因素，但只有10-20%的重度吸烟者会出现症状性疾病。来自动物和人类研究的证据为疾病风险和进展的遗传决定因素提供了支持，但只有一小部分潜在的致病基因被确定。全基因组关联研究是强大的分析工具，可以识别有助于疾病发展和进展的遗传变异。直到最近，这些工具才被用于描述复杂的肺部疾病，如COPD。作为（U 01 HG 004738 -01）慢性阻塞性肺疾病全基因组关联研究的一部分;巴恩斯PI），我们将完成高通量基因分型，其中包括代表多中心NHLBI资助的肺健康研究（LHS）的4，287名欧洲美国参与者的所有已知基因的> 550，000个SNP。对于目前的应用，我们提出了一个广泛的，基于组织的验证方法，利用基因组学和蛋白质检测策略。具体目标1、我们将通过评估来自LTRC的COPD肺组织样本中的全基因组基因表达数据，验证与肺功能下降率和COPD易感性相关的基因中SNP之间最显着的相关性（来自COPD患者（N=30）和具有正常肺功能的非COPD（对照）吸烟者（N=11）的样品，使用Illumina”Sentrix HumanRef-8 Expression BeadChip（Illumina，San Diego，CA）。在特定目标2中，遗传相关性的进一步验证将结合基于蛋白质的免疫印迹和免疫组织化学策略，重点关注与对照标本相比COPD组织中的表达失调和原位定位。我们的研究将为GWAS定义的候选基因使用基于组织的验证策略提供原理证明支持。除了在单独的资助机制下对我们的COPD GWAS的显著发现进行关键验证外，该应用程序的附加价值还包括对40多个LTRC样本的广泛分析，这些样本将随时提供给科学界，以推进阻塞性肺病的其他研究。 (End摘要） 公共卫生相关性： 慢性阻塞性肺病是一个重要的公共卫生问题，更好地了解这种普遍疾病的致病本质对于寻找新的治疗方法至关重要。我们预期将LHS遗传研究中鉴定的遗传变异与从LTRC获得的不同疾病严重程度的充分表征的COPD受试者的肺组织获得的基因表达数据相关联，以鉴定和验证参与COPD发病机制的新候选基因。此外，拟议的研究将为基于互补RNA和蛋白质的方法在验证复杂肺部疾病的遗传结果中的价值提供原理证明支持，该应用的附加价值包括对40个LTRC肺组织样本的广泛分析，这些样本将随时可供科学界使用，以推进阻塞性肺病的其他研究。

DESCRIPTION (provided by applicant): Tissue-based validation of COPD Genetic Studies using Lung Health Study Cohort Chronic obstructive pulmonary disease (COPD) is the fourth leading cause of death in North America, affecting 10-12 million Americans. Although cigarette smoking is the most common environmental risk factor involved in development of COPD, only 10-20% of heavy smokers ever develop symptomatic disease. Evidence from both animal and human studies provides support for genetic determinants for disease risk and progression, yet only a small proportion of the potential causal genes have been identified. Genome wide association studies are powerful analytic tools that can identify genetic variants that contribute to disease development and progression. Only recently have these tools been used to characterize complex lung diseases such as COPD. As part of the (U01HG004738-01) Genome Wide Association Study of Chronic Obstructive Pulmonary Disease; Barnes PI) we will complete high-throughput genotyping which includes > 550,000 SNPs representing all known genes on 4,287 European American participants of the multi-center NHLBI-funded Lung Health Study (LHS). For the current application, we propose a broad, tissue-based validation approach utilizing genomics and protein detection strategies. In Specific Aim 1, we will validate the most significant associations between SNPs in genes associated with rate of decline of lung function and COPD susceptibility by evaluating genome-wide gene expression data in COPD lung tissue specimens from the LTRC (samples from patients with COPD (N=30) and non-COPD (control) smokers with normal lung function (N=11) using the Illumina" Sentrix HumanRef-8 Expression BeadChip (Illumina, San Diego, CA). In Specific Aim 2, further validation of genetic associations will incorporate the protein based strategies of immunoblotting and immunohistochemistry focusing on dysregulated expression and in situ localization in COPD tissue compared with control specimens. Our studies will provide proof-of-principle support for the use of tissue-based validation strategies for GWAS-defined candidate genes. In addition to critical validation of the salient findings of our COPD GWAS under a separate funding mechanism, added value to this application includes the extensive profiling of 40+ LTRC samples that will be readily available to the scientific community to advance other studies of obstructive lung disease. (End of Abstract) PUBLIC HEALTH RELEVANCE: COPD is a significant public health concern, and a better understanding of the pathogenic nature of this pervasive condition is critical to finding novel therapies. We anticipate correlating our genetic variants identified in the LHS genetic studies with gene expression data obtained from lung tissue of well-characterized COPD subjects of varying disease severity obtained from the LTRC in order to identify and validate novel candidate genes involved in the pathogenesis of COPD. In addition, the proposed studies will provide proof-of-principle support for the value of complementary RNA and protein-based approaches in the validation of genetic results for complex lung disorders and added value to this application includes the extensive profiling of 40 LTRC lung tissue samples that will be readily available to the scientific community to advance other studies of obstructive lung disease.