Chemistry and Biology of Deoxyribose Oxidation in DNA

DNA 脱氧核糖氧化的化学和生物学

• 批准号: 8250471
• 负责人: Peter C Dedon
• 金额: $ 37.78万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8250471
• 关键词: Aging; Aldehydes; Area; Benchmarking; Biological; Biological Markers; Biology; Cells; Chemicals; Chemistry; Collaborations; Comparative Study; Complex; DNA; DNA Adducts; DNA Repair; DNA lesion; DNA repair protein; Deoxyribose; Dependence; Development; Dose; Edetic Acid; Electrons; Functional disorder; Funding; Glutathione; Goals; Health; Hydrogen Peroxide; Inflammation; Ionizing radiation; Kinetics; Lesion; Malignant Neoplasms; Metabolic; Metabolism; Methods; Nucleosides; Oxidants; Oxidative Stress; Radiation; Research; Series; Site; Techniques; Toxic effect; Toxicogenetics; Yeasts; analytical method; candidate marker; chemical elimination; comparative; crosslink; human disease; nucleobase; oxidation; particle; repaired; response; ribonolactone; tool

DESCRIPTION (provided by applicant): With the goal of developing biomarkers of oxidative stress, we propose to build on results from the previous funding period by defining the chemical spectrum and biological consequences of deoxyribose oxidation in DNA. DNA oxidation is strongly associated with the pathophysiology of human disease, with a major research focus on nucleobase damage. However, emerging evidence points to deoxyribose oxidation as a critical factor in the toxicity of oxidative stress. We will continue to develop analytical methods to quantify deoxyribose oxidation in isolated DNA and cells exposed to oxidants, and to define the cellular responses to this damage. Aim 1: Develop analytical methods to quantify deoxyribose oxidation products. We propose to continue developing analytical methods to quantify the ribonolactone abasic site from 1'-oxidation, the erythose abasic site from 2'-oxidation, the 3'-oxo species from 3'-oxidation and the nucleoside-5'-aldehyde from 5'-oxidation. This will provide us with a nearly complete set of candidate biomarkers for the various chemistries of deoxyribose oxidation. Aim 2: Comparative analysis of deoxyribose oxidation. In the previous funding period, we initiated comparative studies of deoxyribose oxidation caused by a variety of oxidants to define the determinants of the damage spectrum. We propose to further characterize deoxyribose oxidation caused by ?-radiation, a-particles and FeEDTA, and to include studies of low energy electrons, the most abundant secondary species arising from ionizing radiation. Aim 3: Define the chemistry and biology of deoxyribose oxidation in cells. Using methods developed in Aim 1 and benchmarks determined in Aim 2, we will proceed to study the biological implications of deoxyribose oxidation in cells. There are three goals here: (1) define the biologically relevant spectrum of deoxyribose lesions caused by ?-radiation, a-particles and H2O2 in cells and to identify candidate biomarkers for development in coordination with Aim 4; (2) continue our collaboration with Prof. Bruce Demple to characterize the repair of deoxyribose oxidation products; and (3) pursue our observations of cellular responses to deoxyribose oxidation in yeast made in the previous funding period in terms of the kinetics and dose- dependence of the unique responses to double- and single-strand breaks. Aim 4: Define the metabolic fate of deoxyribose oxidation products. We propose to move our studies forward by defining the metabolic and biotransformational fate of deoxyribose oxidation products. The goal is to identify the chemical transformations that occur with the lesions and the potential utility of the released species as biomarkers. PUBLIC HEALTH RELEVANCE: The broad goal of the proposed studies is to identify candidate molecules for development as biomarkers or indicators of oxidative stress. DNA oxidation is strongly associated with the causative mechanisms of cancer and aging and the bulk of research in this area is focused on damage to the nucleobase components of DNA. However, emerging evidence points to deoxyribose oxidation as a critical factor in the genetic toxicology of oxidative stress and inflammation. We propose to continue our efforts to develop deoxyribose damage products as biomarkers of endogenous and environmentally-induced oxidative stress and inflammation, with the potential for developing predictive and clinically useful tools.

描述（由申请人提供）：为了开发氧化应激的生物标志物，我们建议通过定义DNA中脱氧核糖氧化的化学谱和生物学后果来基于上一个资金时期的结果。 DNA氧化与人类疾病的病理生理密切相关，重点研究核碱基损伤。然而，新兴的证据表明脱氧核糖氧化是氧化应激毒性的关键因素。我们将继续开发分析方法，以量化分离的DNA和暴露于氧化剂的细胞中的脱氧核糖氧化，并定义对这种损伤的细胞反应。目标1：开发分析方法来量化脱氧核糖氧化产物。我们建议继续开发分析方法，以从1'-氧化，2'-氧化，3'-氧化物种和5'-氧化的核苷-5'-5'-醛中量化核糖酮阿无素位点。这将为我们提供几乎完整的候选生物标志物，用于脱氧核糖氧化的各种化学物质。目标2：脱氧核糖氧化的比较分析。在上一个资金期间，我们启动了由多种氧化剂引起的脱氧核糖氧化的比较研究，以定义损伤光谱的决定因素。我们建议进一步表征由辐射，A粒子和饲料引起的脱氧核糖氧化，并包括对低能电子的研究，低能电子是由电离辐射引起的最丰富的二次物种。目标3：定义细胞中脱氧核糖氧化的化学和生物学。使用在AIM 1中开发的方法和在AIM 2中确定的基准，我们将继续研究细胞中脱氧核糖氧化的生物学意义。这里有三个目标：（1）定义细胞中由辐射，A粒子和H2O2引起的脱氧核糖病变的生物学相关光谱，并确定与AIM 4协调开发的候选生物标志物； （2）继续我们与布鲁斯教授的合作，以表征脱氧核糖氧化产物的修复； （3）根据动力学和对双链和单链断裂的独特反应的动力学和剂量依赖性，在上一个资金期间对酵母中对脱氧核糖氧化的反应进行观察。目标4：定义脱氧核糖氧化产物的代谢命运。我们建议通过定义脱氧核糖氧化产物的代谢和生物转化命运来向前发展。目的是确定病变中发生的化学转化以及释放物种作为生物标志物的潜在效用。公共卫生相关性：拟议研究的广泛目标是将候选分子确定为生物标志物或氧化应激指标。 DNA氧化与癌症和衰老的致病机制密切相关，该领域的大部分研究集中在损害DNA的核碱基成分上。然而，新兴的证据表明脱氧核糖氧化是氧化应激和炎症遗传毒理学的关键因素。我们建议继续努力开发脱氧核糖损伤产物，作为内源性和环境诱导的氧化应激和炎症的生物标志物，并有可能开发预测性和临床上有用的工具。