New biomonitoring methodologies to measure DNA adducts in human tissues

测量人体组织中 DNA 加合物的新生物监测方法

• 批准号: 8217315
• 负责人: Robert J. Turesky
• 金额: $ 31.71万
• 依托单位: WADSWORTH CENTER
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-02-01 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8217315
• 关键词: Affect; Animals; Archives; Aristolochia; Aristolochic Acids; Balkan Nephropathy; Balkans; Biological; Biological Assay; Biological Markers; Biological Monitoring; Botanicals; Bread; C3H/He Mouse; Cancer Patient; Carboxylic Acids; Carcinogens; Chemical Models; Chinese Herbs; Chromatography; Chronic Kidney Failure; Clinical; Country; Croatia; DNA; DNA Adduction; DNA Adducts; DNA Repair Enzymes; Data; Diagnosis; Diet; Disease; Epidemiologic Studies; Etiology; Exposure to; Flour; Formalin; Freezing; Funding; Genes; Genetic Predisposition to Disease; Goals; Herb; Human; Incidence; Individual; Infusion procedures; Ingestion; Internet; Ions; Kidney; Kidney Diseases; Kidney Failure; Laboratories; Lesion; Link; Malignant Neoplasms; Mass Fragmentography; Mass Spectrum Analysis; Measurement; Measures; Methodology; Methods; Modeling; Molecular; Molecular Toxicology; Mus; Mutation; Nephrotoxic; PO-1; Patients; Population Study; Public Health; Radioactivity; Renal Tissue; Renal carcinoma; Research; Retrieval; Risk; Role; Rural; Safety; Sampling; Screening procedure; Seeds; Source; Specimen; Staging; Structure; Study Subject; Syndrome; TP53 gene; Techniques; Time; Tissue Sample; Tissues; Toxic effect; Toxicogenomics; Translational Research; United States; United States Food and Drug Administration; United States National Institutes of Health; Xenobiotic Metabolism; adduct; base; cancer risk; carcinogenicity; case control; chemical carcinogen; cost effective; design; exposed human population; global health; human tissue; kidney cortex; liquid chromatography mass spectrometry; mass spectrometer; nano-electrospray; nephrotoxicity; novel; phenanthrene; population based; programs; public health relevance; research study; rural area; tissue fixing; toxicant

DESCRIPTION (provided by applicant): The objective of this proposal is to develop new biomonitoring methodologies designed to measure DNA adducts in tissues of humans exposed to chemical carcinogens. Our chemical model for this project is structurally related nitro-phenanthrene carboxylic acids, a class of human carcinogens and nephrotoxicants that include aristolochic acids (AA). These compounds are universally present in herbs of the genus Aristolochia, used for medicinal purposes throughout the world for 2000 years. The nephrotoxicity and carcinogenicity of herbs containing AA are very well documented. Epidemiologic studies reveal AA to be the causal agent for the clinical syndromes known as Chinese herb and Balkan endemic nephropathies. In the latter, exposure to AA involves ingestion of bread prepared from flour contaminated with seeds of Aristolochia clematitis. Due to their toxicities, importation of traditional Chinese herbs containing AA are banned in some, but not all countries. Despite the Food and Drug Administration's warnings concerning the safety of botanical remedies containing AA, these herbs are still widely distributed in the United States via the Internet, and the incidence of chronic renal failure and urothelial cancer worldwide attributed to exposure to AA remains very high. Recently, we provided unequivocal evidence for the presence of AA-DNA adducts in the renal cortex of patients affected by BEN, using a novel, multi-stage ion trap mass spectrometry (MSn) method. Quantitative MS methods are essential for measuring DNA adduct biomarkers of this devastating and uniformally fatal disease. The biomonitoring methods will be used in translational research studies conducted in Balkan countries where the residents have dietary exposure to AA. A critical technological advance in DNA adduct screening methodologies will be achieved by extending the analysis of AA-DNA adducts from freshly frozen tissue samples to archived, formalin-fixed renal tissues, an untapped but rich source of material for toxico- logical research. Finally, a novel screening method will be established, employing an automated chip-based infusion nano-electrospray tandem MS method. This technique will provide a rapid throughput and cost- effective method to screen for DNA adducts in population-based studies. Traditional herbal remedies containing carcinogenic AA are used worldwide and are a global health problem. Recent epidemiological studies have linked herbs containing AA with renal failure and cancer. Rapid and quantitative analytical MS data to screen for AA-DNA adducts are needed to assess the exposure and the causal role of AA in nephropathy and upper urothelial cancer risk. The AA-DNA adducts also serve as critical biomarkers in studies of genetic susceptibility to Balkan endemic nephropathy and its associated upper urothelial cancer. The novel biomonitoring techniques established in this application can be appllied to examine the role of other chemical carcinogens in the etiology of human cancer. PUBLIC HEALTH RELEVANCE: Novel mass spectrometry biomonitoring methods will be established to measure DNA adducts of aristolochic acids (AA), potent nephrotoxicants and carcinogens found in herbs of the genus Aristolochia, which have been used for medicinal purposes world-wide. These novel screening techniques will be employed to measure AA-DNA adducts in human populations exposed to AA and diagnosed with upper urothelial cancer.

描述（由申请人提供）：本提案的目的是开发新的生物监测方法，旨在测量暴露于化学致癌物的人体组织中的 DNA 加合物。我们该项目的化学模型是结构相关的硝基菲羧酸，这是一类人类致癌物和肾毒物，其中包括马兜铃酸 (AA)。这些化合物普遍存在于马兜铃属草本植物中，在世界各地用于药用已有 2000 年历史。含有 AA 的草药的肾毒性和致癌性已有充分记录。流行病学研究表明，AA 是中草药肾病和巴尔干地方性肾病等临床综合征的致病因素。在后者中，接触 AA 涉及​​摄入由被马兜铃种子污染的面粉制成的面包。由于其毒性，一些国家（但并非所有国家）禁止进口含有 AA 的中药材。尽管美国食品和药物管理局对含有 AA 的植物药物的安全性发出警告，但这些草药仍然通过互联网在美国广泛传播，全球范围内因接触 AA 导致的慢性肾衰竭和尿路上皮癌的发病率仍然很高。 最近，我们使用一种新型多级离子阱质谱 (MSn) 方法，提供了明确的证据，证明 BEN 患者肾皮质中存在 AA-DNA 加合物。定量 MS 方法对于测量这种毁灭性且致命疾病的 DNA 加合物生物标志物至关重要。生物监测方法将用于在居民饮食中接触AA的巴尔干国家进行的转化研究。 DNA 加合物筛选方法的关键技术进步将通过将 AA-DNA 加合物的分析范围从新鲜冷冻的组织样本扩展到存档的福尔马林固定的肾组织来实现，肾组织是毒理学研究尚未开发但丰富的材料来源。最后，将建立一种新颖的筛选方法，采用基于自动化芯片的输注纳米电喷雾串联质谱方法。该技术将为基于人群的研究中筛选 DNA 加合物提供快速通量和经济有效的方法。 含有致癌AA的传统草药在全世界范围内使用，是一个全球性的健康问题。最近的流行病学研究已将含有 AA 的草药与肾衰竭和癌症联系起来。需要快速定量分析 MS 数据来筛选 AA-DNA 加合物，以评估 AA 的暴露及其在肾病和上尿路上皮癌风险中的因果作用。 AA-DNA 加合物还可作为巴尔干地方性肾病及其相关上尿路上皮癌遗传易感性研究的关键生物标志物。本申请中建立的新型生物监测技术可用于检查其他化学致癌物在人类癌症病因学中的作用。 公共健康相关性：将建立新的质谱生物监测方法来测量马兜铃酸 (AA) 的 DNA 加合物、马兜铃属草药中发现的强效肾毒物和致癌物质，这些物质已在世界范围内用于药用目的。这些新颖的筛查技术将用于测量暴露于 AA 并被诊断患有上尿路上皮癌的人群中的 AA-DNA 加合物。