Identification and in vitro experimental investigation of missense SNPs implicate

错义 SNP 的鉴定和体外实验研究

• 批准号: 8366038
• 负责人: OSNAT HERZBERG
• 金额: $ 28.88万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8366038
• 关键词: Affect; Affinity; Alzheimer' s Disease; Amino Acid Substitution; Asthma; Binding; Cell Surface Receptors; Communities; Complex; Computer Simulation; Computing Methodologies; Crohn' s disease; Data; Databases; Development; Differential Scanning Calorimetry; Disease; Disease Association; Down-Regulation; Drug Delivery Systems; Environmental Risk Factor; Evaluation; Genetic Variation; Genome; Genomics; Goals; Hereditary Disease; In Vitro; Individual; Inflammatory Bowel Diseases; Investigation; Knowledge; Methods; Microarray Analysis; Molecular; Mutate; Nature; Play; Predisposition; Process; Property; Proteins; Resources; Rheumatoid Arthritis; Risk; Role; Structure; Surface Plasmon Resonance; Techniques; Thermodynamics; Time; Ultracentrifugation; Variant; Work; disorder risk; genetic variant; genome wide association study; human disease; in vivo; insight; macrophage; mutant; protein function; research study; trait; web interface

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.

描述（由申请人提供）：最近的基因组广泛关联研究导致对多个基因组基因座的可靠鉴定，其中常见SNP的存在与许多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本质的新见解，并对主要常见人类疾病的机制进行了特定的理解。引导对这些基本机制的见解并不是微不足道的，需要开发和应用一系列计算和实验技术。分子过程构成了SNP会影响疾病风险的多级机制的关键部分，并且对其贡献的充分理解对于有效研究更高级别的PATHWA和子系统影响以及对潜在药物靶标的评估至关重要。该项目的重点是错义SNP（导致蛋白质中氨基酸取代的）在疾病机制中的作用。现在很明显，这类SNP在常见疾病机制中起着重要作用。我们专注于与克罗恩病风险增加相关的基因座，那里的关联研究在鉴定疾病相关的基因座方面特别成功。先前开发的计算方法用于确定这些SNP中的哪些有望对相应蛋白质的体内分子功能产生很大的影响，因此最有可能参与疾病机制。目前，25个基因座中有39种蛋白质具有高撞击错义SNP。该分析的结果与其他可用信息结合使用，以优先考虑蛋白质进行实验研究。为了确定对蛋白质功能的精确影响，我们将克隆，表达和纯化许多高优先蛋白及其SNP变体，并研究体外特性，特定的结构稳定性以及与适当结合伴侣的相互作用。我们已经证明了这种策略对于巨噬细胞刺激因子（MSP）的力量，发现通过确定与细胞表面受体的结合受损，这意味着降低了巨噬细胞活性的调节，从而携带了克罗恩病的候选机制SNP。实验性工作的结果以及所有克罗恩病基因座的计算建模和注释将通过在线数据库和注释设施提供，以便可以最大程度地利用结果。 公共卫生相关性：新数据首次可靠地建立了个体之间的遗传变异与对许多常见人类疾病的敏感性之间的许多关联。这些数据为研究疾病潜在机制的研究开辟了道路，从而开发了新疗法。该项目将使用计算和实验方法来确定克罗恩（Crohn's）（一种炎症性肠病）的蛋白质水平机制。