Genetic Factors Governing Inter-individual Variation to Oxidative Stress Response

控制氧化应激反应个体间差异的遗传因素

• 批准号: 8820067
• 负责人: Nicholas Eli Banovich
• 金额: $ 4.31万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-03-01 至 2016-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8820067
• 关键词: 21 year old; African American; Biological Assay; Biological Markers; CD4 Positive T Lymphocytes; Caucasians; Cell Culture Techniques; Cells; DNA Damage; Data; Disease; Etiology; Gene Expression; Genetic; Genetic Variation; Genetic study; Genomic DNA; Genotype; Glutathione Disulfide; Guanine; Health; Helper-Inducer T-Lymphocyte; Hour; Immunoglobulin Variable Region; Individual; Link; Longevity; Maps; Measures; Mediating; Messenger RNA; Oxidative Stress; Oxygen; Pathway interactions; Physiological; Population; Quantitative Trait Loci; RNA Sequences; Reduced Glutathione; Reporter Genes; Sampling; Series; Single Nucleotide Polymorphism; Site-Directed Mutagenesis; Specific qualifier value; Time; Variant; age related; base; bead chip; biological adaptation to stress; caucasian American; genome-wide; improved; interest; research study; response; sex; trait; transcriptome sequencing

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.

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密切相关，并且是长寿的决定因素。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与年龄相关的疾病和生理衰退的理解。在这里，我提出了一系列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，在目标 1 中，我将从 70 名白人和 70 名非裔美国人身上纯化 CD4+ T 细胞。我将这些细胞暴露在氧化应激下 72 小时，方法是在升高的氧气水平 (20%) 下培养它们，同时将来自同一个体的对照细胞保持在生理 (5%) 的氧气水平。在这期间的指定时间点，我会测量氧化型谷胱甘肽与还原型谷胱甘肽的比率（氧化应激的指标）；氧化鸟嘌呤与未修饰鸟嘌呤的比例 鸟嘌呤（氧化应激引起的 DNA 损伤的指标）；以及处理细胞和对照细胞中全基因组 mRNA 丰度。在目标 2 中，我将从目标 1 中使用的所有样本收集基因型，并绘制数量性状位点 (QTL)，包括表达 QTL，以解释氧化应激反应的变化。然后，目标 3 将使用报告基因检测来验证大约 30 个假定的 eQTL。