Kidney DNA Adductomics

肾DNA加成组学

• 批准号: 10631192
• 负责人: Silvia Balbo
• 金额: $ 44.31万
• 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/10631192
• 关键词: Aflatoxins; Alcohols; Algorithms; Applications Grants; Aristolochic Acids; Asia; Automation; Bioinformatics; Biological Assay; Cancer Etiology; Cancer Patient; Carcinogens; Categories; Characteristics; Chemical Agents; Chemical Exposure; Chemicals; Complex; Computer software; DNA; DNA Adduction; 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; Hazardous Chemicals; Herbal Medicine; Human; Human Genome; Individual; Induced Mutation; Inflammation; Investigation; Kidney; 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 Induction; 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; 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; nano; 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.

项目摘要 人类经常暴露于环境中的危险化学物质，这可能会损害DNA和 导致癌症。但是，在过去的30年中，DNA加合物的测量已大大携带 通过监视单个或最多几个DNA加合物，数据提供了有限的信息 癌症的病因。建立强大的技术以筛选广泛的技术是至关重要的。 DNA加合物的范围，其中一些可以与癌症驱动基因突变有关。我们应用的目标 tion是为了推进我们的质谱（MS）加合体方法来筛选一系列DNA加合物 在单个测定中用危险化学物质形成，并开发算法以进行自动分析。我们会 策略高分辨率准确的质谱法（HRAMS）来检测DNA加合物，通过提取特征 DNA加合物的特性光谱特征通过多阶段（MSN）扫描进行碎片化。这些 MSN扫描方法可以检测到广泛的人体组织中不同结构的DNA加合物 暴露。在此提案中，我们将在受控的暴露下对啮齿动物的MS技术进行优化 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发展 软件和信息学工具，用于全面表征基因组的DNA加合物。 在AIM 1中，啮齿动物的肾脏将产生DNA加合曲线，在暴露于有毒物质之后 在烟草，环境，饮食中发现，来自促氧化剂和内源性产生的电力。面板 这些暴露产生的DNA加合曲线的特征是未靶向数据依赖性（DDA） 和独立于数据的获取（DIA），采用纳米流液色谱和多阶段MSN 使用Orbitrap MS进行扫描。在AIM 2中，将构建DNA加合物数据库和生物信息学工具 将通过提取质量光谱而为对DNA加合物的无偏和自动检测而创建 DNA加合物的特征。生物信息学工具将确定与不同类别形成的加合物的模式 基于不同类型的肾脏有毒物质并进行系统比较和鉴定DNA 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，以检查不同类别的DNA 人类的加合物。在AIM 3中，我们将测试我们的技术以筛查肾癌患者的活检并进行探索 烟草暴露是该癌症的少数公认危险因素之一。 提出的用于DNA加合物分子分析的技术可与其他靶器官和细胞相提并论 类型并可以帮助识别诱导DNA损伤和癌症风险的化学药物和生活方式因素。 鉴定DNA损害剂可以导致减轻或防止暴露于危险的策略 对公共卫生产生巨大影响的化学物质。