MtDNA damage as a biomarker for environmental mitochondrial toxicity

线粒体 DNA 损伤作为环境线粒体毒性的生物标志物

• 批准号: 8623989
• 负责人: J Timothy Greenamyre
• 金额: $ 44.62万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-19 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8623989
• 关键词: Accounting; Address; Affect; Age; Agriculture; Behavioral; Biochemical; Biological; Biological Assay; Biological Markers; Blood; Blood specimen; Bolus Infusion; Case-Control Studies; Chemicals; Chronic; Complex; Cost Savings; DNA; DNA Damage; DNA Library; DNA lesion; Defect; Dose; Environmental Risk Factor; Epidemiologic Studies; Epidemiologist; Epidemiology; Etiology; Exposure to; Funding; Gender; Genetic; Genomic DNA; Health; Herbicides; Human; Injection of therapeutic agent; Institutes; Laboratories; Lead; Mitochondria; Mitochondrial DNA; Modeling; Mus; Nerve Degeneration; Nested Case-Control Study; Occupational; Occupational Exposure; Oxidation-Reduction; Oxidative Stress; Paraquat; Parents; Parkinson Disease; Participant; Pesticides; Polymerase; Positioning Attribute; Principal Investigator; Rattus; Reaction Time; Reactive Oxygen Species; Reporting; Research Design; Risk; Risk Factors; Rodent; Rotenone; Sampling; Smoking; Solvents; Specimen; Spouses; Subgroup; Technology; Testing; Tetrachloroethylene; Toxic Environmental Substances; Toxic effect; Toxin; Translational Research; Trichloroethylene; United States; Universities; Work; base; cohort; dopamine system; experience; exposed human population; farmer; gene environment interaction; human disease; illness length; inhibitor/antagonist; innovation; novel; parent grant; pesticide exposure; public health relevance

DESCRIPTION (provided by applicant): The Greenamyre laboratory (University of Pittsburgh) was recently funded to examine mitochondrial DNA (mtDNA) damage as a potential biomarker of exposure to environmental mitochondrial toxins. The PCR-based assay employed is extremely sensitive, quantitative and highly reproducible - and it detects any type of damage that slows polymerase progression. The parent grant, upon which the current application is based, is experimental and examines various aspects of mtDNA damage in rats exposed to mitochondrial toxins. The basic premise is that mitochondrial toxins, by generating reactive oxygen species (ROS), will cause various forms of DNA lesions in the mitochondria. Parkinson's disease (PD), affecting about 1 million people in the United States, appears to involve both genetic and environmental factors - and systemic mitochondrial defects have been strongly implicated. The best characterized environmental risk factor for PD is occupational exposure to pesticides, many of which are mitochondrial inhibitors. In this regard, the current application engages two of the epidemiologists who were instrumental in identifying specific pesticides as bona fide risk factors for PD. The new collaborators, Caroline Tanner and Sam Goldman (Parkinson's Institute, Sunnyvale, CA), showed that the risk of PD more than doubled in farmers using rotenone or paraquat, which are both ROS-generating mitochondrial inhibitors. Additionally, they reported that exposure to the solvent, trichloroethylene (TCE), also increases the risk of PD. In rats, TCE exposure is associated with inhibition of mitochondrial complex I. Importantly, existing genomic DNA samples are available from the epidemiological studies of Tanner and Goldman. Because (i) PD is associated with mitochondrial defects and exposures to known mitochondrial toxins, and (ii) even subclinical exposure of rats to a mitochondrial toxin produces robust mtDNA damage in blood, they propose specific aims to address the following questions: Does paraquat and/or TCE exposure cause mtDNA damage? Is human exposure to rotenone, paraquat or TCE associated with mtDNA damage? Is sporadic PD, with or without exposure to rotenone, paraquat or TCE associated with mtDNA damage? The current proposal builds upon the strengths of the parent application, with its innovative and extremely sensitive assay for mtDNA damage, and establishes a bridge from experimental work in rodents (Greenamyre) to translational research in the human epidemiology of Parkinson's disease (Tanner & Goldman). Using existing technology and staff, and banked DNA specimens, they will leverage current assets in an effort to develop a biomarker for human exposures to certain environmental toxicants (e.g., rotenone, paraquat & TCE) and, possibly, for Parkinson's disease. As a result, the proposed work can be done with dramatic cost savings and efficiency compared to a new study. Moreover, the principal investigators have a unique opportunity to test our assay as a biomarker for PD in two independent epidemiological cohorts. Given their combined experience and expertise, they believe they are uniquely positioned to assess this potential biomarker.

描述（由申请人提供）：最近资助了Greenamyre实验室（匹兹堡大学），以检查线粒体DNA（mtDNA）损害，作为潜在暴露于环境线粒体毒素的生物标志物。所采用的基于PCR的测定非常敏感，定量且高度可再现 - 它检测到减慢聚合酶进展的任何类型的损伤。当前应用所基于的父授予是实验性的，并检查了暴露于线粒体毒素的大鼠中mtDNA损伤的各个方面。基本的前提是，线粒体毒素通过产生活性氧（ROS）将在线粒体中引起各种形式的DNA病变。在美国影响约100万人的帕金森氏病（PD）似乎涉及遗传和环境因素 - 以及全身性线粒体缺陷。 PD的最佳特征环境风险因素是职业接触农药，其中许多是线粒体抑制剂。在这方面，当前的应用程序与两位流行病学家介入，他们有助于将特定农药识别为PD的真正风险因素。新的合作者Caroline Tanner和Sam Goldman（加利福尼亚州桑尼维尔的帕金森研究所）表明，使用Rotenone或Paraquat的农民中PD的风险增加了一倍以上，它们都是玫瑰果生成的线粒体抑制剂。此外，他们报告说，暴露于溶剂，三氯乙烯（TCE）也会增加PD的风险。在大鼠中，TCE暴露与线粒体复合物的抑制I有关，重要的是，现有的基因组DNA样品可以从Tanner和Goldman的流行病学研究中获得。因为（i）PD与线粒体缺陷和暴露于已知的线粒体毒素，以及（ii）大鼠在线粒体毒素中的亚临床暴露会在血液中产生强大的mtDNA损害，他们提出特定的目的，以解决以下问题：paraquat and paraquat and paraquat and或tce and tece and tece and tece and tece and tece and或tce and/tecdna损害吗？人类暴露于烤面包酮，paraquat或TCE是否与mtDNA损害相关？是否有散发性PD，是否有或不暴露于烤面包酮，paraquat或tce与mtDNA损伤相关的情况下？当前的提案建立在父母应用的优势基础上，其创新性和极其敏感的测定法对mtDNA损害造成损害，并建立了从啮齿动物（Greenamyre）（Greenamyre）的实验性工作到帕金森氏病（Tanner＆Goldman）人类流行病学的转化研究的桥梁。他们将使用现有的技术和员工以及存储的DNA标本，他们将利用流动资产来开发人类对某些环境毒物（例如Rotenone，Paraquat＆TCE）的暴露生物标志物，并可能为帕金森氏病。结果，与一项新研究相比，可以通过巨大的成本和效率来完成拟议的工作。此外，首席研究人员有一个独特的机会，可以在两个独立的流行病学队列中测试我们作为PD的生物标志物的测定法。鉴于他们的综合经验和专业知识，他们认为自己在评估这种潜在的生物标志物方面处于独特状态。