Genomic instability, susceptibility to oxidative stress and cellular senescence

基因组不稳定、对氧化应激和细胞衰老的易感性

基本信息

批准号：
9304443
负责人：
Marcus Stanley Cooke
金额：
$ 36.63万
依托单位：
FLORIDA INTERNATIONAL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-05-01 至 2020-08-06
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9304443
关键词：
Address Aging Aging-Related Process Automobile Driving Binding Binding Sites Biological Cardiovascular Diseases Cardiovascular system Cell Aging Cell Cycle Stage Cell model Cell physiology Cells Cellular Metabolic Process ChIP-seq Chemicals Coupled DNA DNA Damage DNA Repair Data Deoxyguanosine Development Disease Environment Event Exhibits Exons Exposure to Functional disorder Generations Genes Genome Genomic Instability Genomic Segment Goals Health Human Immunoprecipitation Intervention Introns Link Location Malignant Neoplasms Maps Measurement Methods Modeling Neurodegenerative Disorders Normal Cell Nuclear Nucleic Acid Regulatory Sequences Organism Oxidative Stress Oxidative Stress Induction Pathologic Pattern Play Predisposition Process Production Proteins Pyrimidine Dimers Research Role Site Stem cells Stress Students Techniques adult stem cell anthropogenesis base chromosomal location environmental stressor human disease insight knock-down next generation sequencing oxidation oxidative DNA damage repaired senescence spatiotemporal stressor

项目摘要

Project Summary / Abstract All living organisms are continually exposed to a variety of environmental stressors, be they anthropogenic or natural in origin. Many stressors share a common toxic mechanism that can have detrimental effects on the cell (e.g., generation of highly reactive chemical species). A proportion of these chemical species, evade the cell's defenses and damage cellular components including DNA, causing oxidation. Measurement of global genome levels of oxidatively damaged DNA, implicates environmental stressors in major human health issues (e.g., cancer, aging, cardiovascular, and neurodegenerative diseases). However, current associations between DNA damage and disease are based upon crude assessments of global genome damage, which provide limited mechanistic information on how damage leads to disease. Furthermore, DNA damage is not uniformly distributed across the genome; accumulation, or persistence, of damage in regions of the genome vital to the functioning of the cell will have downstream consequences. We propose, that the role of DNA damage in disease can only be understood by examination of damage in the context of its location. We currently lack information concerning how the cell maintains baseline levels of oxidatively damaged DNA, and its spatio-temporal distribution across the genome. This is fundamental to our understanding of how the cell responds to damage and targets regions for prioritized repair. The objective of this study is to identify regions of the genome with increased susceptibility to the formation, or slow repair, of oxidatively damaged DNA that play a functional role in senescence. This study will engage students in independent, meritorious research, strengthening the institutional research environment. Aim 1. To identify susceptible regions of the genome for oxidative stress-induced damage. A. Characterize the distribution of basal levels 8-oxodG, and its repair surveillance. Patterns of DNA damage and repair will be identified using DDIP-seq for 8-oxodG, and ChIP-seq for hOGG1 in an aging stem cell model. B. Develop a model to interrogate the factors influencing the distribution of DNA damage and repair and predict downstream effects. DNA damage and repair will be mapped to specific gene sequences, introns/exons, regulatory sequences, and chromosomal locations. Identifying regions of the genome that may have functional consequences in cellular dysfunction and ROS-induced senescence. C. Assess the role of nuclear organization on damage and repair. FISH will be utilized to form spatio-temporal topological maps of DNA damage and repair across the genomic regions identified in 1A and 1B. Aim 2. To determine the mechanisms linking increased susceptibility to oxidative stress to senescence. Examine the effect of increased endogenous ROS on the targeting of DNA damage and repair. We hypothesize that increased ROS alters the distribution of damage and repair driving the disease process. Using the above approaches, in our aging stem cell model with increased endogenous ROS production. We hypothesize that any new regions identified will represent candidates for having a role in triggering senescence.

项目概要/摘要所有生物体都不断暴露于各种环境压力源，无论是人为的还是起源自然。许多压力源都有一个共同的毒性机制，可能对细胞产生有害影响（例如，产生高活性化学物质）。这些化学物质的一部分，逃避细胞的防御并损害包括 DNA 在内的细胞成分，导致氧化。全球基因组测量氧化损伤 DNA 的水平，表明环境压力因素与重大人类健康问题有关（例如，癌症、衰老、心血管疾病和神经退行性疾病）。然而，目前 DNA 之间的关联损害和疾病是基于对全球基因组损害的粗略评估，该评估提供的信息有限有关损伤如何导致疾病的机制信息。此外，DNA损伤并不是均匀分布的跨越基因组；对功能至关重要的基因组区域损伤的积累或持续该细胞将产生下游后果。我们认为，DNA 损伤在疾病中的作用只能是通过检查损害所在位置来理解。我们目前缺乏有关的信息细胞如何维持氧化损伤 DNA 的基线水平及其时空分布基因组。这是我们理解细胞如何响应损伤和目标区域的基础以进行优先修复。这项研究的目的是确定基因组中易感性增加的区域氧化损伤 DNA 的形成或缓慢修复，在衰老过程中发挥功能作用。这项研究将让学生进行独立、有价值的研究，加强机构研究环境。目标 1. 确定基因组中氧化应激引起的损伤的易感区域。 A. 表征基础水平 8-oxodG 的分布及其修复监测。 DNA 损伤的模式在衰老干细胞模型中，将使用 DDIP-seq 检测 8-oxodG 和 ChIP-seq 检测 hOGG1 来识别修复。 B. 开发模型来探究影响 DNA 损伤分布的因素以及修复和预测下游效应。 DNA 损伤和修复将被映射到特定的基因序列、内含子/外显子、调控序列和染色体位置。识别基因组中可能具有功能的区域细胞功能障碍和 ROS 诱导的衰老的后果。 C. 评估核组织对损伤和修复的作用。 FISH将被用来形成时空 1A 和 1B 中确定的基因组区域 DNA 损伤和修复的拓扑图。目标 2. 确定氧化应激易感性增加与衰老之间的联系机制。检查内源性 ROS 增加对 DNA 损伤和修复的影响。我们假设 ROS 的增加改变了驱动疾病过程的损伤和修复的分布。使用上面的方法，在我们的衰老干细胞模型中，内源性 ROS 产量增加。我们假设任何确定的新区域将代表在触发衰老中发挥作用的候选区域。