Genomics of COPD

项目总结(见说明)： 全基因组关联研究已经确定了与COPD易感性的重复关联，包括15号染色体(CHRNA3/5，IREB2)、HHIP和FAM13A。多项研究还检查了COPD患者的全基因组基因表达谱。基因表达谱分析可以识别新的COPD基因，并帮助确定这些GWAS发现的基因影响COPD的途径。来自生物学和功能验证研究的初步数据表明，IREB2是通过基因表达谱和GWAs在人类COPD中发现的基因之一，可能通过影响自噬来影响COPD。这一方法指出了COPD的一种新途径，即自噬。 自噬已被证明在常见疾病中发挥作用，我们的小组已经证明了自噬和细胞凋亡在肺气肿中的潜在相关性。在本项目中，我们将测试两个假设： (1)通过对人COPD肺组织和小鼠模型的基因表达谱分析，找出与COPD相关的新基因以及与IREB2、HHIP和FAM13A相关的基因，以阐明这些基因在COPD中的作用途径。(2)通过基因表达谱鉴定的COPD易感基因IREB2和GWAs在香烟烟雾诱导下调节自噬过程，进而促进下游细胞凋亡促进肺气肿的发生。我们将解决以下问题 具体目标：(1)人类基因表达谱：我们将在人类肺组织样本中进行微阵列基因表达谱分析，以确定COPD患者和对照组之间差异表达的基因。将验证患有和不患有COPD的吸烟者的呼吸道上皮细胞和肺泡巨噬细胞中的基因表达，以确定基因表达的组织特异性。(2)小鼠模型的基因表达：我们将对两个野生型菌株进行肺组织微阵列基因表达谱的研究。 在香烟烟雾暴露前后的3种基因敲除小鼠模型(Ireb2-/-，HHIP-/-，Fam13a-/-)中，以及在气液界面培养的呼吸道上皮细胞模型中，研究了对香烟烟雾诱导的肺气肿的不同易感性，以确定这些动物中与COPD发展相关的基因。(3)Ireb2-/-和自噬：我们将确定Ireb2在实验性香烟烟雾诱导的肺气肿易感性中的功能作用。在本项目结束时，我们将确定COPD的新基因，并加深我们对HHIP、FAM13A和IREB2在COPD中的作用的理解，特别是对自噬和凋亡途径的影响。本项目中对人类和小鼠基因表达的评估将分别补充项目1和项目3中的遗传学和表观遗传学方法。这些数据集的整合将扩大我们对COPD功能基因组学的了解。

PROJECT SUMMARY (See instructions): Genome-wide association studies (GWAS) have identified replicated associations with COPD susceptibility, including the chromosome 15 locus {CHRNA3/5, IREB2), HHIP, and FAM13A. Multiple studies have also examined genome-wide gene expression profiles in patients with COPD. Gene expression profiling can identify novel COPD genes and help define the pathways by which these GWAS-discovered genes affect COPD. Preliminary data from biological and functional validation studies suggests that IREB2, one such gene identified through gene expression profiling and GWAS in human COPD, may influence COPD through effects on autophagy. This approach has pointed to a novel pathway in COPD, namely autophagy. Autophagy has been shown to play a role in common diseases, and our group has demonstrated the potential relevance of autophagy and apoptosis in emphysema. In this project, we will test two hypotheses: (1) Gene expression profiling in human COPD lung tissue and mouse models will identify novel genes for COPD and genes whose expression is correlated with IREB2, HHIP, and FAM13A, in order to elucidate the pathways of these genes' actions in COPD. (2) IREB2, a COPD susceptibility gene identified through gene expression profiling and GWAS, when induced by cigarette smoke regulates the autophagic process, which facilitates downstream apoptosis promoting the development of emphysema. We will address the following Specific Aims: (1) Human Gene Expression Profiling: We will perform micro-array gene expression profiling in human lung tissue samples to identify genes that are differentially expressed between COPD cases and controls. Gene expression will be validated in airway epithelial cells and alveolar macrophages from former smokers with and without COPD to determine tissue specificity of gene expression. (2) Gene Expression in Murine Models: We will perform micro-array gene expression profiling in lung tissue in 2 wild-type strains with differing susceptibility to cigarette smoke-induced emphysema and in 3 knock-out mouse models { Ireb2-/-, Hhip-/- ,Fam13a-/-) before and after cigarette smoke exposure and in a cell model of airway epithelial cells cultured at an air-liquid interface to identify genes involved in COPD development in these animals. (3) Ireb2-/- and autophagy: We will determine the functional role of Ireb2 in susceptibility to experimental cigarette smoke-induced emphysema. At the conclusion of this project, we will have identified novel genes for COPD and improved our understanding of the roles of HHIP, FAM13A, and IREB2 in COPD, with specific attention to the effects on autophagy and apoptosis pathways. The assessment of human and murine gene expression in this project will complement the genetics and epigenetics methods in Projects 1 and 3, respectively. Integration of these datasets will expand our knowledge of the functional genomics of COPD.