Oxidative DNA Damage and CpG Mutagenesis

DNA 氧化损伤和 CpG 突变

基本信息

批准号：
8106959
负责人：
Yinsheng Wang
金额：
$ 23.64万
依托单位：
UNIVERSITY OF CALIFORNIA RIVERSIDE
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-07-01 至 2016-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8106959
关键词：
8,5&apos -cyclo-2&apos -deoxyadenosine 8,5&apos -cyclo-2&apos -deoxyguanosine Address Affect Biological Biological Assay Biological Markers Biology Bypass Cells Chemicals Chemistry Copper CpG dinucleotide Cytosine DNA DNA Damage DNA Repair DNA biosynthesis DNA lesion DNA-Directed DNA Polymerase Data Deoxyguanosine Development Epigenetic Process Escherichia coli Exposure to Fenton&apos s reagent Fibroblasts Funding Gene Expression Regulation Generations Genomics Goals Grant Guanine Hepatolenticular Degeneration Human Hydrogen Peroxide In Vitro Individual Ions Knowledge Lead Lesion Liver Malignant Neoplasms Mammalian Cell Measures Methods Methylation Modeling Molecular Biology Molecular Profiling Monitor Mutagenesis Mutation Nucleotide Excision Repair Oligodeoxyribonucleotides Organic Chemistry Outcome Paper Patients Photochemistry Play Production Property Proteins Publishing Purines Rattus Reaction Reactive Oxygen Species Reagent Research Risk Factors Role Route Shuttle Vectors Site Solid Source Techniques Technology Testing Thermodynamics Tissues Transition Elements ascorbate base brain tissue bromocytosine crosslink cytotoxic exposed human population human disease improved in vivo innovation insight interdisciplinary approach methyl radical nervous system disorder novel oxidative DNA damage purine repaired tumor

项目摘要

DESCRIPTION (provided by applicant): Reactive oxygen species (ROS) can be induced by both endogenous and exogenous sources and they can result in DNA damage. We hypothesize that the ROS-induced bulky DNA lesions, owing to their difficulty in being repaired and their miscoding potential, may contribute significantly to the ubiquitously observed mutations at CpG dinucleotide site. To test this hypothesis, we will take an interdisciplinary approach, which encompasses synthetic organic chemistry, bioanalytical chemistry and molecular biology, to investigate the formation of these lesions in vitro and in vivo and to assess how they are interpreted by DNA replication and repair machineries in human cells. We organize the proposed research into three specific aims: In Aim 1, we will assess quantitatively the formation of bulky lesions in DNA upon exposure to Fenton reagents and investigate how cytosine methylation affects the formation of 8,5'-cyclo-2'-deoxyguanosine at CpG site. We will also quantify these lesions formed in tissues of a rat model of Wilson's disease, which arises from excessive accumulation of copper ion in the liver. In Aim 2, we will investigate the cytotoxic and mutagenic properties of these lesions in mammalian cells by using our newly developed shuttle vector technology. In Aim 3, we will examine how the ROS-induced bulky DNA lesions are repaired in human cells. The significance of the proposed research lies in that it will provide important insights into the implications of these lesions in the development of human diseases including cancer and neurological disorders. Additionally, the results from the proposed research may reveal potential risk factors for developing human diseases and lead to the discovery of new molecular signatures for monitoring the human exposure toward ROS. The proposed research is highly innovative because it involves the use of a multi-pronged approach to tackle an important, yet largely overlooked biological problem. PUBLIC HEALTH RELEVANCE: The focus of this proposal is to assess the formation and biological consequences bulky DNA lesions induced by reactive oxygen species (ROS). The outcome of the proposed research will improve significantly our understanding of the implications of ROS exposure in the development of cancer and neurological disorders and may lead to the discovery of novel biomarkers for monitoring the human exposure toward ROS.

描述（由申请人提供）：活性氧（ROS）可以由内源性和外源性来源诱导，它们可以导致DNA损伤。我们假设ROS诱导的笨重的DNA病变，由于其难以修复和误导潜力，可能会对CPG Dinucleotide部位普遍观察到的突变产生重大贡献。为了检验这一假设，我们将采用一种跨学科方法，其中包括合成有机化学，生物分析化学和分子生物学，以研究体外和体内这些病变的形成，并评估人类细胞中DNA复制和修复机器的解释方式。我们将拟议的研究组织为三个特定的目的：在AIM 1中，我们将在暴露于Fenton试剂后定量评估DNA中笨重的病变的形成，并研究胞嘧啶甲基化如何影响CPG部位的8,5'- cyclo-2'-甲氧基因鸟氨酸的形成。我们还将量化在威尔逊氏病大鼠模型的组织中形成的这些病变，该病变是由于肝脏中铜离子过度积累而产生的。在AIM 2中，我们将通过使用新开发的班车矢量技术研究这些病变在哺乳动物细胞中这些病变的细胞毒性和诱变特性。在AIM 3中，我们将研究如何在人类细胞中修复ROS诱导的大量DNA病变。拟议研究的意义在于，它将为这些病变在包括癌症和神经系统疾病在内的人类疾病发展中的影响提供重要的见解。此外，拟议研究的结果可能揭示了发展人类疾病的潜在风险因素，并导致发现新的分子特征来监测人类对ROS的暴露。拟议的研究具有很高的创新性，因为它涉及使用多管齐下的方法来解决一种重要但很大程度上被忽视的生物学问题。公共卫生相关性：该提案的重点是评估由活性氧（ROS）诱导的大量DNA病变的形成和生物学后果。拟议研究的结果将显着改善我们对ROS暴露在癌症和神经系统疾病发展中的影响的理解，并可能导致发现新型生物标志物来监测人类对ROS的暴露。