DJ-1 in Astrocyte-Mediated Neuroprotection Against Complex I Inhibitors

DJ-1 在星形胶质细胞介导的针对复合物 I 抑制剂的神经保护中

基本信息

批准号：
8476788
负责人：
J Timothy Greenamyre
金额：
$ 33.41万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-15 至 2016-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8476788
关键词：
1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine 1-Methyl-4-phenylpyridinium Affect Analytical Chemistry Animal Model Astrocytes Behavioral Biochemical Brain Brain region Cell Culture Techniques Coculture Techniques Complex Data Disease Epidemiologic Studies Exposure to Genetic Glial Fibrillary Acidic Protein Goals Human In Vitro Link Mediating Methods Midbrain structure Mitochondria Modeling Mus Mutation Nerve Degeneration Neuraxis Neurodegenerative Disorders Neurons Occupational Exposure PARK7 protein Parkinson Disease Parkinsonian Disorders Pesticides Process Publishing Rattus Reporting Research Research Project Grants Respiratory Chain Risk Risk Factors Rotenone Subfamily lentivirinae Substantia nigra structure System Testing Therapeutic Toxic Environmental Substances Transgenic Mice Work astrocyte mediated neuroprotection cell type dopaminergic neuron environmental pesticide exposure fenazaquin fenpyroximate gene therapy in vivo inhibitor/antagonist intraperitoneal killings knock-down neuron loss neuroprotection neurotoxic novel novel therapeutics pars compacta pesticide exposure promoter release factor translational approach

项目摘要

DESCRIPTION (provided by applicant): Parkinson's disease (PD) is characterized by progressive neuronal loss in multiple brain regions. Particularly affected is the substantia nigra pars compacta (SNpc) of the midbrain. In PD, this region shows degeneration of dopaminergic neurons and deficient mitochondrial respiratory chain complex I activity. 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), a complex I inhibitor, has caused human parkinsonism by selectively killing SNpc dopaminergic neurons. Several pesticides, including rotenone, also selectively inhibit complex I and cause SNpc dopaminergic neurodegeneration in animal models. Human epidemiological studies have now linked pesticide exposure to an increased risk for PD. One recent study implicated occupational exposure to rotenone. It is therefore plausible that environmental toxicant (pesticide) exposure causes the complex I deficiency seen in PD, and that this mechanism is central to disease initiation and/or progression. Our research goal is to identify natural protective strategies employed by the brain against neurodegeneration induced by complex I-inhibiting environmental toxicants. Ultimately, we hope to exploit these mechanisms to develop novel disease-modifying therapies against PD. Our recent work has identified DJ-1 over-expression in astrocytes as a neuroprotective mechanism against complex I-inhibiting pesticides in vitro. This process appears to involve astrocyte-released factors, and may be particularly relevant to PD because (i) reactive astrocytes over-express DJ-1 in sporadic PD and (ii) mutations that eliminate DJ-1 expression cause familial PD. Thus, we hypothesize that astrocytic DJ-1 over-expression in the human brain may represent a natural neuroprotective attempt against PD. If true, this process may be targetable for augmentation as disease-modifying therapy. To model this possibility in the intact brain, we will assess the capacity of astrocytic DJ-1 over-expression to reduce MPTP-induced SNpc dopaminergic neurodegeneration in transgenic mice (Aim 1). This will be studied using behavioral, immunohistochemical, and biochemical analyses in novel mice, recently developed in our lab, that over- express DJ-1 selectively in astrocytes under control of the glial fibrillary acidic protein promoter. We will also assess the capacity of lentivirus-mediated astrocytic DJ-1 over-expression to perform similarly against rotenone in rats (Aim 2). In each case, we hypothesize that astrocytic DJ-1 over-expression will augment astrocyte-mediated neuroprotection against complex I inhibition. Aim 2 will also test an experimental translational gene therapy approach against pesticide-induced neurodegeneration. In our final Aim we will use analytical chemistry methods to identify the DJ-1-modulated, astrocyte-released soluble factor(s) that carry the neuroprotective activity in our cell culture model. These factors, or the mechanisms they employ, may also generate novel therapeutic strategies against pesticide-induced neurodegeneration and PD.

描述（由申请人提供）：帕金森病（PD）的特征是多个大脑区域进行性神经元丧失。特别受影响的是中脑的黑质致密部 (SNpc)。在 PD 中，该区域显示多巴胺能神经元退化和线粒体呼吸链复合物 I 活性缺陷。 1-甲基-4-苯基-1,2,3,6-四氢吡啶 (MPTP) 是一种复合物 I 抑制剂，通过选择性杀死 SNpc 多巴胺能神经元而引起人类帕金森病。包括鱼藤酮在内的几种农药也会选择性抑制复合物 I，并在动物模型中引起 SNpc 多巴胺能神经变性。人类流行病学研究现已将接触农药与帕金森病风险增加联系起来。最近的一项研究表明职业接触鱼藤酮。因此，环境毒物（农药）暴露可能会导致帕金森病中出现的复合物 I 缺乏，并且这种机制对于疾病的发生和/或进展至关重要。我们的研究目标是确定大脑针对复杂 I 抑制性环境毒物引起的神经变性所采用的自然保护策略。最终，我们希望利用这些机制来开发针对帕金森病的新型疾病缓解疗法。我们最近的工作已确定星形胶质细胞中 DJ-1 的过度表达是体外对抗复合物 I 抑制性农药的神经保护机制。这一过程似乎涉及星形胶质细胞释放的因子，并且可能与 PD 特别相关，因为 (i) 反应性星形胶质细胞在散发性 PD 中过度表达 DJ-1，以及 (ii) 消除 DJ-1 表达的突变导致家族性 PD。因此，我们假设人脑中星形胶质细胞 DJ-1 的过度表达可能代表了针对 PD 的自然神经保护尝试。如果属实，这个过程可能可以作为疾病缓解疗法来增强。为了在完整大脑中模拟这种可能性，我们将评估星形胶质细胞 DJ-1 过度表达减少转基因小鼠中 MPTP 诱导的 SNpc 多巴胺能神经变性的能力（目标 1）。我们将在我们实验室最近开发的新型小鼠中使用行为、免疫组织化学和生化分析来研究这一点，这些小鼠在神经胶质纤维酸性蛋白启动子的控制下在星形胶质细胞中选择性过度表达 DJ-1。我们还将评估慢病毒介导的星形胶质细胞 DJ-1 过表达对大鼠中鱼藤酮的类似作用（目标 2）。在每种情况下，我们假设星形胶质细胞 DJ-1 过度表达将增强星形胶质细胞介导的针对复合物 I 抑制的神经保护作用。目标 2 还将测试针对农药引起的神经变性的实验性转化基因治疗方法。在我们的最终目标中，我们将使用分析化学方法来鉴定 DJ-1 调节的星形胶质细胞释放的可溶性因子，这些因子在我们的细胞培养模型中具有神经保护活性。这些因素或其采用的机制也可能产生针对农药引起的神经变性和帕金森病的新治疗策略。