Environmental Mitochondrial Toxicants Cause LRRK2 Activation in Parkinson's Disease

环境线粒体毒物导致帕金森病中 LRRK2 激活

• 批准号: 10213962
• 负责人: Briana De Miranda
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10213962
• 关键词: Acute; Adult; Affect; Animal Model; Antibodies; Antigen Presentation; Attenuated; Autophagocytosis; Biological Assay; CRISPR/Cas technology; Cell Line; Cell membrane; Cell surface; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; DNA Sequence Alteration; Development; Disease; Dose; Embryo; Environment; Environmental Pollution; Environmental Risk Factor; Etiology; Exposure to; Foundations; Functional disorder; Funding; Gene Mutation; Genes; Genetic Engineering; Genetic Predisposition to Disease; Genus Hippocampus; Goals; Herbicides; Human; Idiopathic Parkinson Disease; Immune; Impairment; Individual; Industrialization; Inherited; Ketoglutarate Dehydrogenase Complex; LRRK2 gene; Lead; Ligation; Link; Measures; Mentors; Mentorship; Mitochondria; Mitochondrial Proteins; Modeling; Movement Disorders; Mutation; Nerve Degeneration; Neurons; PARK8 gene; Paraquat; Parkinson Disease; Pathogenesis; Pathogenicity; Pathologic; Pathology; Penetrance; Pesticides; Phase; Phosphorylation; Phosphotransferases; Population; Positioning Attribute; Proteins; Rattus; Reporting; Research; Risk; Risk Factors; Rodent Model; Role; Rotenone; Soil; Solvents; Susceptibility Gene; Technology; Therapeutic; Toxic Environmental Substances; Toxic effect; Toxicant exposure; Toxicology; Training; Trichloroethylene; Vesicle; aged; alpha synuclein; base; cellular pathology; combat; cytotoxic; disorder risk; dopaminergic neuron; drinking water; gene environment interaction; ground water; in vivo; inhibitor/antagonist; innovation; kidney cell; kinase inhibitor; mitochondrial dysfunction; mutant; neuroinflammation; neuron loss; novel; novel therapeutics; organic contaminant; programs; protein transport; response; targeted treatment; tenure track; toxicant; trafficking

Abstract The cause of idiopathic Parkinson’s disease (PD) remains unknown, however, significant evidence suggests that interaction between genetic susceptibility and environmental factors is the predominant etiology of PD. Environmental toxicants that cause mitochondrial dysfunction, such as the organic pesticide rotenone, and the common herbicide paraquat, are associated with elevated PD risk (OR 2.5, 95% CI: 0.1.3-4.7; OR 2.5, 95% CI: 1.4-4.7; respectively). A heavily used industrial solvent, trichloroethylene (TCE), also causes mitochondrial toxicity, and is the most frequently reported organic contaminant found in US groundwater. TCE exposure is linked to the development of PD (OR 6.1, 95% CI: 1.2-3.3), and rodent models of TCE exposure display dopamine neuron degeneration from the nigrostriatal tract. Recent evidence from our lab indicates that rotenone (ROT), paraquat (PQ), and TCE interact with PD susceptibility genes, notably, causing the activation of LRRK2 in wildtype (WT) human embryonic kidney (HEK) cells, which could be blocked by a selective LRRK2 inhibitor (GNE-7915). As LRRK2 is the most commonly inherited mutation associated with familial PD, this evidence suggests that a gene-environment interaction exists between LRRK2 and mitochondrial toxicants. Functionally, LRRK2 activation leads to multiple downstream cellular pathologies, such as disruption of vesicular trafficking, deficits in autophagy, the phosphorylation of α-synuclein, and neuroinflammation; all of which are mechanisms hypothesized to precede dopamine neuron degeneration in PD. The basis of this proposal is to investigate LRRK2 activation and pre-degenerative mechanisms in dopamine neurons caused by environmental mitochondrial toxicants. To achieve this, we will pursue the following specific aims: Aim 1 (K99) will build a foundation to identify if LRRK2 activity is induced in WT neurons by environmental mitochondrial toxicants, and if LRRK2 mutations exacerbate this pathology following mitochondrial dysfunction. Aim 2 (R00) will further characterize LRRK2 activation in an animal model of TCE exposure, and determine whether LRRK2 inhibition is protective against TCE. Aim 3 (R00) will characterize another pre-degenerative mechanism influenced by environment mitochondrial toxicants and LRRK2, mitochondrial antigen presentation (MitAP). MitAP involves the trafficking of mitochondrial proteins from the inner lumen to cell surface MHC molecules, causing the selective killing of dopamine neurons by immune cells. We have measured MitAP in dopamine neurons following a single exposure to ROT in rats, suggesting this is an early response to mitochondrial toxicity. We will identify whether ROT, PQ, and TCE induce MitAP in dopaminergic neurons. As LRRK2 activity affects vesicular trafficking, we propose that LRRK2 mutations in N27 cell lines influences MitAP in response to mitochondrial toxicant exposure, and treatment with a LRRK2 inhibitor may be protective against MitAP in vivo. These aims will provide innovative evidence for LRRK2 activation by environmental factors that contributes to dopaminergic neuron degeneration. Collectively, this proposal may lead to new therapeutic treatment avenues for idiopathic and inherited PD.

抽象的 特发性帕金森氏病（PD）的原因仍然未知，但是，大量证据表明， 遗传敏感性和环境因素之间的相互作用是PD的主要病因。 引起线粒体功能障碍的环境有毒物质，例如有机农用烤烤鱼酮和 普通除草剂帕拉夸酸与PD风险升高有关（OR 2.5，95％CI：0.1.3-4.7; OR 2.5，95％CI： 1.4-4.7;分别）。大量使用的工业偿付能力，三氯乙烯（TCE）也会引起线粒体 毒性，是在美国地下水中发现的最常见的有机污染物。 TCE暴露是 与PD的开发（OR 6.1，95％CI：1.2-3.3）和TCE暴露显示模型有关 斑纹纹状体的多巴胺神经元变性。我们实验室的最新证据表明鱼藤酮 （腐烂），paraquat（PQ）和TCE与PD易感基因相互作用，特别是引起LRRK2的激活 在野生型（WT）人类胚胎肾（HEK）细胞中，可以被选择性LRRK2抑制剂阻止 （GNE-7915）。由于LRRK2是与家族性PD相关的最常见的突变，因此 表明LRRK2和线粒体有毒物质之间存在基因环境相互作用。在功能上 LRRK2激活导致多种下游细胞病理，例如囊泡贩运的破坏， 自噬，α-核蛋白的磷酸化和神经炎症的缺陷；所有这些都是机制 假设在PD中以多巴胺神经元的变性为前。该提议的基础是调查 由环境引起的多巴胺神经元中的LRRK2激活和分离前机制 线粒体有毒物质。为了实现这一目标，我们将追求以下具体目标：AIM 1（K99）将建立一个 基础，确定环境线粒体有毒物质在WT神经元中是否诱导LRRK2活性，并且 如果LRRK2突变加剧了线粒体功能障碍后这种病理。 AIM 2（R00）将进一步 在TCE暴露的动物模型中表征LRRK2激活，并确定LRRK2抑制是否为 防止TCE。 AIM 3（R00）将表征另一种受影响的预苯前机制 环境线粒体毒物和LRRK2，线粒体抗原表现（MITAP）。 mitap涉及 从内腔到细胞表面MHC分子的线粒体蛋白运输，导致选择性 免疫细胞杀死多巴胺神经元。我们已经测量了一个单一的多巴胺神经元中的mitap 暴露于大鼠腐烂，这表明这是对线粒体毒性的早期反应。我们将确定是否 腐烂，PQ和TCE影响多巴胺能神经元中的MITAP。由于LRRK2活性会影响囊泡贩运，我们 提议N27细胞系中的LRRK2突变会影响线粒体毒物暴露的MITAP， 用LRRK2抑制剂治疗可以在体内保护MITAP。这些目标将提供创新的 LRRK2激活的证据是由环境因素激活的，导致多巴胺能神经元变性。 总的来说，该提案可能会导致针对特发性和遗传PD的新的治疗途径。