Genetic Factors Governing Inter-individual Variation to Oxidative Stress Response

描述（由申请人提供）：我建议测量个体和群体之间氧化应激反应的变化，并确定与这种变化相关的遗传位点。氧化应激与年龄相关疾病的病因学密切相关，并作为长寿的决定因素。介导氧化应激反应的途径已被广泛研究，但我们对这些途径功能的个体差异仍然知之甚少。了解控制个体氧化应激反应变化的因素可能会提高我们对年龄相关疾病和生理衰退的理解。在此，我提出了一系列的实验和统计分析来研究氧化应激反应变异的遗传基础。具体来说，在Aim 1中，我将从70名白种人和70名非裔美国人中纯化CD4+ T细胞。我将把这些细胞置于氧化应激72小时，方法是将它们培养在高氧（20%）水平下，同时将来自同一个体的对照细胞保持在生理氧水平（5%）下。在此期间的指定时间点，我将测量氧化谷胱甘肽与还原性谷胱甘肽的比例（氧化应激指标）；氧化鸟嘌呤与未修饰鸟嘌呤的比值

DESCRIPTION (provided by applicant): I propose to measure variation in oxidative stress response across individuals and populations, and to identify the genetic loci that are associated with such variation. Oxidative stress has been strongly implicated in the etiology of age-related disease and as a determinant of longevity. Pathways that mediate oxidative stress response have been extensively studied, but we still know relatively little about individual variation in th function of these pathways. Understanding the factors that govern variation in oxidative stress response across individuals may improve our understanding of age-related disease and physiological decline. Here, I propose a series of experiments and statistical analyses to study the genetic basis of variation in oxidative stress response. Specifically, in Aim 1, I will purify CD4+ T cells from 70 Caucasians and 70 African Americans. I will expose these cells to oxidative stress for 72 hours by culturing them in elevated (20%) oxygen levels, while maintaining control cells from the same individuals in physiological (5%) oxygen levels. At specified time points during this period, I will measure the ratio of oxidized glutathione to reduced glutathione (an indicator of oxidative stress); the ratio of oxidized guanine to unmodified guanine (an indicator of oxidative stress-induced DNA damage); and genome-wide mRNA abundance, in both treated and control cells. In Aim 2, I will collect genotypes from all samples used in aim 1 and map quantitative trait loci (QTLs), including expression QTLs, that explain variation in oxidative stress response. Reporter gene assays will then be used in Aim 3 to validate approximately 30 of the putative eQTLs.