Kidney DNA Adductomics

肾DNA加成组学

• 批准号: 10424477
• 负责人: Silvia Balbo
• 金额: $ 44.3万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-07 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10424477
• 关键词: Aflatoxins; Alcohols; Algorithms; Applications Grants; Aristolochic Acids; Asia; Automation; Bioinformatics; Biological Assay; Cancer Etiology; Cancer Patient; Carcinogens; Characteristics; Chemical Agents; Chemical Exposure; Chemicals; Complex; Computer software; DNA; DNA Adducts; DNA Damage; DNA Repair; DNA analysis; Data; Data Analyses; Databases; Detection; Developed Countries; Development; Diet; Environment; Enzymes; Etiology; Europe; Exposure to; Gender; Genes; Genetic Polymorphism; Genome; Goals; Gold; Hazardous Chemicals; Herbal Medicine; Human; Human Genome; Individual; Induced Mutation; Inflammation; Investigation; Kidney; Lead; Leukocytes; Link; Liquid Chromatography; Malignant Neoplasms; Malignant neoplasm of liver; Mammary gland; Mass Fragmentography; Mass Spectrum Analysis; Measurement; Measures; Meat; Medicine; Methods; Molecular Analysis; Monitor; Mutation; Organ; Oxidative Stress; Partner in relationship; Pattern; Pesticides; Pollution; Population; Predisposition; Process; Prostate; Proteomics; Public Health; Rattus; Reactive Oxygen Species; Renal Cell Carcinoma; Renal carcinoma; Research Design; Resolution; Risk Assessment; Risk Factors; Rodent; Role; Sampling; Scanning; Smoker; Software Tools; Specificity; Specimen; Structure; Techniques; Technology; Testing; Tissues; Tobacco; Tobacco smoke; United States; Xenobiotic Metabolism; adduct; analytical tool; automated algorithm; automated analysis; base; bioinformatics tool; biomarker discovery; cancer risk; cell type; chemical genetics; cohort; cooking; environmental mutagens; environmental tobacco smoke; epidemiology study; human tissue; improved; informatics tool; kidney biopsy; lifestyle factors; metabolomics; non-smoker; novel; prevent; software development; tobacco exposure; tool; toxicant

PROJECT SUMMARY Humans are frequently exposed to hazardous chemicals in the environment, which can damage DNA and lead to cancer. However, over the past 30 years, the measurements of DNA adducts have been largely carried out by monitoring a single or up to several DNA adducts, and the data have provided limited information about causative agents of cancer. There is a crucial unmet need to establish a robust technology to screen for a broad range of DNA adducts, some of which can be linked to mutations in cancer driver genes. The goal of our applica- tion is to advance our mass spectrometry (MS) adductomics methods to screen for an array of DNA adducts formed with hazardous chemicals in a single assay and develop algorithms for automated analysis. We will em- ploy high-resolution accurate mass spectrometry (HRAMS) to detect DNA adducts by extracting the charac- teristic spectral features of DNA adducts as they undergo fragmentation by the multistage (MSn) scanning. These MSn scanning approaches can detect DNA adducts of diverse structures in human tissues over a wide range of exposures. In this proposal, we will optimize our MS technology in rodents, under controlled exposures to an array of chemicals thought to contribute to renal cancer, and we will create a mass spectral database and develop software and informatics tools for comprehensive characterization of DNA adducts of the genome. In Aim 1, DNA adduct profiles will be generated in the kidney of rodents, following exposures to toxicants found in tobacco, the environment, diet, from pro-oxidants, and electrophiles produced endogenously. Panels of DNA adduct profiles resulting from these exposures will be characterized by untargeted data-dependent (DDA) and data-independent acquisition (DIA), employing nanoflow liquid chromatography and multistage MSn scanning with the Orbitrap MS. In Aim 2, a DNA adduct database will be constructed, and bioinformatics tools will be created for the unbiased and automated detection of DNA adducts by extraction of the mass spectral features of DNA adducts. The bioinformatics tools will identify patterns of adducts formed with different classes of renal toxicants and conduct systematic comparison and identification of DNA adducts based on different types of exposures and the mechanism of DNA damage. The DIA and DDA scanning methods, the DNA adduct database, and bioinformatics technologies will serve as templates to examine for different classes of DNA adducts in humans. In Aim 3, we will test our technologies to screen biopsies of renal cancer patients and explore tobacco exposure, one of the few recognized risk factors for this cancer. The proposed technology for molecular analysis of DNA adducts is amenable to other target organs and cell types and can help identify chemical agents and lifestyle factors that induce DNA damage and cancer risk. Identification of DNA-damaging agents can lead to strategies to mitigate or prevent exposures to hazardous chemicals with a dramatic impact on public health.

项目概要 人类经常接触环境中的危险化学物质，这些化学物质会损害 DNA 和 导致癌症。然而，在过去的30年里，DNA加合物的测量基本上已经进行了 通过监测单个或最多几个 DNA 加合物来得出结论，并且数据提供的信息有限 癌症的病原体。目前迫切需要建立一种强大的技术来筛选广泛的 一系列 DNA 加合物，其中一些可能与癌症驱动基因的突变有关。我们的应用程序的目标 我们的目标是推进我们的质谱 (MS) 加合物组学方法来筛选一系列 DNA 加合物 在一次测定中用危险化学品形成并开发自动分析算法。我们将- 利用高分辨率精确质谱 (HRAMS) 通过提取特征来检测 DNA 加合物 DNA 加合物通过多级 (MSn) 扫描进行碎片化时的立体光谱特征。这些 MSn 扫描方法可以检测人体组织中多种结构的 DNA 加合物 曝光。在本提案中，我们将在啮齿动物中优化我们的 MS 技术，在受控暴露于 一系列被认为会导致肾癌的化学物质，我们将创建一个质谱数据库并开发 用于全面表征基因组 DNA 加合物的软件和信息学工具。 在目标 1 中，接触有毒物质后，啮齿类动物的肾脏中会产生 DNA 加合物谱 存在于烟草、环境、饮食、内源产生的促氧化剂和亲电子物质中。面板 由这些暴露产生的 DNA 加合物谱的特征将是非靶向数据依赖 (DDA) 和数据独立采集 (DIA)，采用纳流液相色谱和多级 MSn 使用 Orbitrap MS 进行扫描。目标2将构建DNA加合物数据库，并利用生物信息学工具 将创建用于通过提取质谱来公正且自动地检测 DNA 加合物 DNA加合物的特点。生物信息学工具将识别不同类别形成的加合物的模式 肾毒物的研究，并根据不同类型对DNA加合物进行系统比较和鉴定 暴露和 DNA 损伤机制。 DIA 和 DDA 扫描方法，DNA 加合物 数据库和生物信息学技术将作为模板来检查不同类别的 DNA 人类体内的加合物。在目标 3 中，我们将测试我们的技术来筛查肾癌患者的活检并探索 接触烟草是为数不多的公认的导致这种癌症的危险因素之一。 所提出的 DNA 加合物分子分析技术适用于其他靶器官和细胞 类型，并可以帮助识别诱发 DNA 损伤和癌症风险的化学制剂和生活方式因素。 识别 DNA 损伤剂可以制定减轻或防止接触危险物质的策略 对公众健康产生巨大影响的化学品。