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)：格林阿梅尔实验室(匹兹堡大学)最近获得资助，以检查线粒体DNA(MtDNA)损伤，作为暴露于环境线粒体毒素的潜在生物标记物。采用的基于聚合酶链式反应的分析是极其敏感、定量和高度重复性的-它可以检测到任何类型的损伤，减缓聚合酶的进展。目前的申请所基于的父母拨款是实验性的，它检查了暴露在线粒体毒素下的大鼠线粒体DNA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物种(ROS)，会在线粒体中造成各种形式的DNA损伤。帕金森氏病(PD)在美国影响着大约100万人，似乎涉及遗传和环境因素-系统性线粒体缺陷被强烈牵连。帕金森病的最具特征的环境风险因素是职业暴露于农药，其中许多是线粒体抑制剂。在这方面，目前的申请涉及两名流行病学家，他们帮助确定特定的杀虫剂是帕金森病的真正危险因素。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，使用鱼藤酮或百草枯的农民患帕金森病的风险增加了一倍以上，这两种药物都是产生ROS的线粒体抑制剂。此外，他们报告说，暴露在三氯乙烯(TCE)溶剂中也会增加帕金森病的风险。在大鼠中，TCE暴露与线粒体复合体I的抑制有关。重要的是，现有的基因组DNA样本可以从Tanner和Goldman的流行病学研究中获得。由于(I)PD与线粒体缺陷和暴露于已知的线粒体毒素有关，以及(Ii)即使是亚临床暴露于线粒体毒素的大鼠也会在血液中产生强大的mtDNA损伤，他们提出了具体的目标来解决以下问题：百草枯和/或三氯乙烯暴露是否会导致mtDNA损伤？人类接触鱼藤酮、百草枯或三氯乙烯是否与线粒体DNA损伤有关？有或没有鱼藤酮、百草枯或三氯乙烯暴露的散发性帕金森病是否与线粒体DNA损伤有关？目前的建议建立在亲本应用程序的优势之上，其创新和极其敏感的线粒体DNA损伤分析，并建立了一座桥梁，从啮齿类动物的实验工作(Greenamre)到帕金森氏病人类流行病学的转化研究(Tanner&Goldman)。利用现有的技术和人员，以及储存的DNA样本，他们将利用现有的资产，努力开发一种生物标记物，用于人类暴露于某些环境毒物(如鱼藤酮、百草枯和三氯乙烯)，并可能用于帕金森氏症。因此，与一项新的研究相比，拟议的工作可以显著节省成本和提高效率。此外，主要研究人员有一个独特的机会，在两个独立的流行病学队列中测试我们作为帕金森病生物标志物的检测方法。考虑到他们的经验和专业知识，他们相信他们在评估这一潜在的生物标记物方面处于独特的地位。