DNA Cross-linking by diepoxybutane

二环氧丁烷 DNA 交联

• 批准号: 8197537
• 负责人: NATALIA Y TRETYAKOVA
• 金额: $ 22.11万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-01 至 2013-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8197537
• 关键词: 1,3-Butadiene; 3,4-epoxy-1-butene; Adenine; Alkylating Agents; Automobile Exhaust; Base Pairing; Biological; Butadiene; Butylene Glycols; Bypass; Carcinogens; Characteristics; Chemicals; Cities; Complex Mixtures; Confidential Information; Cyclization; DNA; DNA Adducts; DNA Alkylation; DNA Repair; DNA Structure; DNA alkyltransferase; DNA lesion; DNA repair protein; Data; Detection; Epidemiologic Studies; Ethylene Oxide; Excision; Exposure to; Funding; Gene Mutation; Goals; Guanine; Health; Hematologic Neoplasms; High Pressure Liquid Chromatography; Human; Hydrolysis; Inhalation Exposure; Instruction; Laboratories; Laboratory Animals; Laboratory Rat; Laboratory mice; Language; Lesion; Lung Neoplasms; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Metabolic Activation; Methodology; Methods; Minnesota; Mission; Molecular; Molecular Conformation; Molecular Structure; Mus; Mutagenesis; Mutation; Organism; Pathway interactions; Physiological; Play; Point Mutation; Positioning Attribute; Preparation; Prevalence; Progress Reports; Property; Proteins; Proteomics; Public Health; Publishing; Rattus; Reaction; Research; Research Design; Research Methodology; Risk; Risk Assessment; Role; Site; Site-Directed Mutagenesis; Specificity; Structure; Techniques; Testing; Tissue Extracts; Tissues; Tobacco smoke; Uncertainty; University of Minnesota Cancer Center; Work; adduct; base; cancer risk; carcinogenesis; cigarette smoking; crosslink; cytotoxic; cytotoxicity; design; ds-DNA; erythritol anhydride; exposed human population; human DNA; improved; in vivo; innovation; insight; leukemia; repaired; research study; tool

DESCRIPTION: See instructions. State the application's broad, long-term objectives and specific aims, making reference to the health relatedness of the project (i.e., relevance to the mission of the agency). Describe concisely the research design and methods for achieving these goals. Describe the rationale and techniques you will use to pursue these goals. In addition, in two or three sentences, describe in plain, lay language the relevance of this research to public health. If the application is funded, this description, as is, will become public information. Therefore, do not include proprietary/confidential information. DO NOT EXCEED THE SPACE PROVIDED. 1,2,3,4-diepoxybutane (DEB) is a genotoxic intermediate produced upon the metabolic activation of 1,3- butadiene (BD), a known human carcinogen produced industrially and found in automobile exhaust and in cigarette smoke. DEB is the most mutagenic and cytotoxic metabolite of BD and is likely to play an important role in BD-induced carcinogenesis. The presence of two oxirane groups within the molecular structure of DEB allows it to form DNA-DNA cross-links by consecutively alkylating two adjacent nucleobases in a DNA duplex. In addition, DEB can form potentially promutagenic exocyclic lesions by alkylating two sites of the same DNA base. The long-range goal of our research is to establish the molecular mechanisms by which bifunctional alkylating agents elicit their biological effects. The objective of this project is to identify specific DNA lesions responsible for the genotoxic effects of DEB and BD. The central hypothesis of this research is that DEB forms DNA-DNA cross-links and exocyclic adducts that accumulate in target tissues, contributing to the observed carcinogenic and mutagenic properties of BD. Our proposed studies will improve the current understanding of the mechanisms of mutagenesis and cytotoxicity resulting from BD exposure by providing key information about bifunctional DEB-DNA adducts, including their formation in vivo following exposure to BD, their effects on DNA structure, mispairing characteristics, and cellular repair. We will be pursuing the following four Specific Aims: 1. Quantify bifunctional DEB-DNA lesions in vivo following inhalation exposure to 1,3-butadiene. A sensitive and specific mass spectrometry-based methodology will be used to analyze DNA-DNA cross- links and exocyclic DEB adducts in DNA extracted from tissues of mice and rats exposed to BD. 2. Determine the effects of bifunctional DEB-DNA adducts on DNA duplex structure and replication. Structural analyses by NMR will be performed to analyze adduct conformations in double stranded DNA, while site specific mutagenesis experiments will determine translesion bypass efficiencies and mutational properties of each DEB-DNA adduct. 3. Analyze the repair of bifunctional DEB-DNA adducts. We will identify the major DNA repair mechanisms responsible for the removal of DEB-DNA adducts and analyze the relationships between adduct conformations and repair efficiency. 4. Characterize DNA-protein cross-linking by DEB. A combination of proteomics and immunological detection will be used to investigate DNA-protein cross-linking by DEB as an additional pathway to cytotoxicity and mutagenesis of BD. Collectively, these studies will identify bifunctional DNA adducts responsible for the biological activity of BD and afford new insights into the mechanisms of its mutagenicity and cytotoxicity, reducing the uncertainty in cancer risk assessment for human exposure to BD. PERFORMANCE SITE(S) (organization, city, state) The Cancer Center, University of Minnesota Minneapolis, Minnesota

描述：参见说明。说明该应用程序的广泛、长期目标和具体目标，并参考以下方面的健康相关性 项目（即与机构使命的相关性）。简明地描述实现这些目标的研究设计和方法。描述 您将用于实现这些目标的基本原理和技术。 此外，用两到三个句子，用通俗易懂的语言描述这项研究与公共卫生的相关性。如果申请获得资助，这 描述将按原样成为公共信息。因此，请勿包含专有/机密信息。不要超出空间 假如。 1,2,3,4-二环氧丁烷 (DEB) 是 1,3- 代谢活化时产生的遗传毒性中间体 丁二烯 (BD)，一种工业生产的已知人类致癌物，存在于汽车尾气和 香烟烟雾。 DEB 是 BD 最具致突变性和细胞毒性的代谢物，可能发挥重要作用 BD 诱导的癌发生中的作用。分子结构中存在两个环氧乙烷基团 DEB 可以通过连续烷基化 DNA 中的两个相邻核碱基来形成 DNA-DNA 交联 复式。此外，DEB 可以通过烷基化两个位点来形成潜在的促突变外环损伤。 相同的DNA碱基。我们研究的长期目标是建立分子机制 双功能烷化剂引发其生物效应。该项目的目标是确定具体 DNA 损伤导致 DEB 和 BD 的基因毒性作用。本研究的中心假设是 DEB 形成 DNA-DNA 交联和环外加合物，在靶组织中积累，有助于 观察到的 BD 致癌和致突变特性。我们提出的研究将改善当前的 通过提供了解 BD 暴露引起的突变和细胞毒性的机制 关于双功能 DEB-DNA 加合物的关键信息，包括它们在暴露于 BD，它们对 DNA 结构、错配特征和细胞修复的影响。我们将追求 以下四个具体目标： 1. 定量吸入暴露于 1,3-丁二烯后体内双功能 DEB-DNA 损伤。 将使用灵敏且特异的基于质谱的方法来分析 DNA-DNA 交叉 从暴露于 BD 的小鼠和大鼠组织中提取的 DNA 中存在链接和环外 DEB 加合物。 2. 确定双功能 DEB-DNA 加合物对 DNA 双链体结构和复制的影响。 将通过 NMR 进行结构分析，以分析双链 DNA 中的加合物构象， 而位点特异性诱变实验将确定跨损伤旁路效率和突变 每个 DEB-DNA 加合物的特性。 3. 分析双功能DEB-DNA加合物的修复。我们将确定主要的 DNA 修复机制 负责去除DEB-DNA加合物并分析加合物之间的关系 构象和修复效率。 4. 通过 DEB 表征 DNA-蛋白质交联。蛋白质组学和免疫学的结合 检测将用于研究 DEB 的 DNA-蛋白质交联，作为一种额外的途径 BD 的细胞毒性和诱变。 总的来说，这些研究将鉴定负责 BD 生物活性的双功能 DNA 加合物 并为其致突变性和细胞毒性的机制提供新的见解，减少不确定性 人类接触 BD 的癌症风险评估。 绩效站点（组织、城市、州） 明尼苏达大学癌症中心 明尼苏达州明尼阿波利斯