Identification and in vitro experimental investigation of missense SNPs implicate

描述(由申请人提供)：最近的全基因组关联研究已导致可靠地识别多个基因组座位，其中常见SNP的存在与一些常见人类疾病的风险改变有关。这些结果提供了对复杂性状疾病的一般性质的新见解以及对主要常见人类疾病潜在机制的具体理解的前景。要洞察这些潜在的机制并不是一件容易的事，需要开发和应用一系列计算和实验技术。在SNPs影响疾病风险的多层次机制中，分子过程是关键部分，充分了解它们的作用对于有效研究更高水平的途径和子系统的影响以及评估潜在的药物靶点至关重要。这个项目的重点是错义SNPs(那些导致蛋白质中的氨基酸替换的SNPs)在疾病机制中的作用。现在很清楚，这类SNP在常见疾病机制中发挥着重要作用。我们专注于与克罗恩病风险增加相关的基因座，其中关联研究在识别疾病相关基因座方面特别成功。以前开发的计算方法被用来确定这些SNP中的哪些有望对相应蛋白质的体内分子功能产生较大影响，从而最有可能参与疾病机制。目前，有25个基因座的39种蛋白质存在候选的高影响错义SNPs。这一分析的结果与其他可用信息相结合，为实验研究确定蛋白质的优先顺序。为了确定对蛋白质功能的精确影响，我们将克隆、表达和纯化一些高优先级的蛋白质及其SNP变体，并研究其体外性质，特别是结构稳定性以及与合适的结合伙伴的相互作用。我们已经在巨噬细胞刺激因子(MSP)的情况下证明了这一策略的力量，MSP被发现携带克罗恩病的候选机制SNP，通过建立与细胞表面受体的结合受损，意味着巨噬细胞活动的下调。实验工作的结果以及对所有克罗恩病基因的计算建模和注释将通过在线数据库和注释工具提供，以便最大限度地利用结果。 公共卫生相关性：新数据首次可靠地确定了个体间的遗传变异与一些常见人类疾病的易感性之间的许多联系。这些数据为研究疾病的潜在机制，从而开发新的治疗方法开辟了道路。这个项目将使用计算和实验方法来确定克罗恩病(一种炎症性肠病)致病基因的蛋白质水平机制。

DESCRIPTION (provided by applicant): Recent genome wide association studies have led to the reliable identification of multiple genomic loci where the presence of common SNPs is associated with altered risk of a number of common human diseases. These results offer the prospect of new insights into the nature of complex trait disease in general and specific understanding of the mechanisms underlying major common human diseases. Deriving insight into these underlying mechanisms is not trivial, and requires the development and application of a range of computational and experimental techniques. Molecular processes constitute a key part of the multiple level mechanisms by which SNPs influence disease risk, and a full understanding of their contributions is essential to effective investigation of higher level pathwa and subsystems impact, as well as evaluation of potential drug targets. This project focuses on the role of missense SNPs (those that result in an amino acid substitution in a protein) in disease mechanism. It is now clear that this class of SNP plays a substantial role in common disease mechanisms. We focus on loci associated with increased risk of Crohn's disease, where association studies have been particularly successful in identifying disease relevant loci. Previously developed computational methods are used to identify which of these SNPs are expected to have a large impact on the in vivo molecular function of the corresponding protein, and thus are most likely to be involved in disease mechanism. Currently, there are 39 proteins in 25 loci that have candidate high impact missense SNPs. Results of this analysis are combined with other available information to prioritize proteins for experimental study. In order to determine the precise effect on protein function, we will clone, express and purify a number of the high priority proteins and their SNP variants, and investigate in vitro properties, particularl structural stability and interaction with appropriate binding partners. We have demonstrated the power of this strategy for the case of macrophage stimulating factor (MSP), found to carry a candidate mechanism SNP for Crohn's disease, by establishing that binding to a cell surface receptor is impaired, implying down regulation of macrophage activity. Results of the experimental work together with computational modeling and annotation for all the Crohn's disease loci will made available through an online database and annotation facility, so that the results can be maximally exploited. PUBLIC HEALTH RELEVANCE: New data are for the first time reliably establishing many associations between genetic variation among individuals and susceptibility to a number of common human diseases. These data open the way for investigation of the mechanisms underlying disease and hence the development of new therapies. This project will use computational and experimental methods to determine protein level mechanisms underlying disease loci for Crohn's, an inflammatory bowel disease.