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.

项目摘要 /摘要所有生物体都不断暴露于各种环境压力源，无论是人为的还是天然起源。许多压力源具有一种常见的有毒机制，可能对细胞产生不利影响（例如，高反应性化学物种的产生）。这些化学物种的一部分，逃避了细胞防御和损害细胞成分，包括DNA，导致氧化。全球基因组的测量氧化受损的DNA水平，暗示了主要人类健康问题中的环境压力源（例如，癌症，衰老，心血管和神经退行性疾病）。但是，DNA之间的当前关联损害和疾病是基于对全球基因组损害的粗略评估，这提供了有限的有关损害如何导致疾病的机械信息。此外，DNA损伤并不均匀分布跨基因组；基因组区域损害的积累或持久性对功能至关重要该单元将带来下游后果。我们建议，DNA损害在疾病中的作用只能是通过检查其位置的损坏来理解。我们目前缺乏有关的信息细胞如何维持氧化损坏的DNA的基线水平及其跨时空分布基因组。这是我们对细胞如何应对损害和针对区域的理解至关重要的用于优先修复。这项研究的目的是确定具有提高易感性的基因组区域氧化受损的DNA的形成或缓慢修复，在衰老中起功能的作用。这项研究将吸引学生进行独立的，有功的研究，从而增强机构研究环境。目的1。确定基因组的敏感区域以氧化应激诱导的损伤。 A.表征基底水平8-OXODG的分布及其修复监视。 DNA损伤的模式并将使用DDIP-SEQ用于8-OXODG，在老化的干细胞模型中使用DDIP-Seq进行chip-Seq。 B.开发一个模型来询问影响DNA损伤分布和修复的因素，并预测下游效应。 DNA损伤和修复将映射到特定的基因序列，内含子/外显子，调节序列和染色体位置。识别可能具有功能性的基因组区域细胞功能障碍和ROS诱导的衰老的后果。 C.评估核组织在破坏和维修中的作用。鱼将被用来形成时空在1a和1b中鉴定的基因组区域的DNA损伤和修复的拓扑图。目标2。确定将氧化应激敏感性增加与衰老的敏感性增加的机制。检查内源性ROS增加对DNA损伤和修复的靶向的影响。我们假设 ROS增加了损害的分布和修复的分布，推动了疾病过程。使用以上在我们老化的干细胞模型中，内源性ROS产生增加。我们假设任何人确定的新区域将代表候选人在触发衰老中发挥作用。