Role of LRRK2 in idiopathic Parkinson's disease

LRRK2 在特发性帕金森病中的作用

• 批准号: 10224659
• 负责人: J Timothy Greenamyre
• 金额: $ 34.23万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10224659
• 关键词: Aftercare; Anatomy; Antibodies; Antioxidants; Autopsy; Binding; Biochemical; Biological Assay; Brain; Cells; Co-Immunoprecipitations; Consensus; Dissociation; Gene Proteins; Hippocampus (Brain); Human; Hydrogen Peroxide; Idiopathic Parkinson Disease; In Situ; In Vitro; Individual; LRRK2 gene; Ligation; Mediating; Microglia; Mitochondria; Mitochondrial DNA; Modeling; Mutation; Nerve Degeneration; Neuroglia; Neurons; Oxidative Stress; Parkinson Disease; Pathogenesis; Pathology; Phosphorylation; Phosphotransferases; Physiological; Play; Population; Post-Translational Protein Processing; Proteins; Rattus; Resolution; Risk Factors; Role; Rotenone; Signal Transduction; Specificity; Specimen; Western Blotting; alpha synuclein; base; brain tissue; case control; cell type; disease-causing mutation; dorsal motor nucleus; genomic locus; human tissue; immunocytochemistry; in vivo; inhibitor/antagonist; kinase inhibitor; knock-down; lymphoblast; mutant; mutation carrier; nigrostriatal system; novel; olfactory bulb; overexpression; oxidation; virtual

Mutations in LRRK2 cause familial Parkinson disease (PD) and, in some populations, may account for up to 40% of all cases3. The LRRK2 gene locus also contains a risk factor for `idiopathic' PD (iPD); however, the role of LRRK2 in typical iPD is not clear. Furthermore, the relationship of LRRK2 to other genes and proteins associated with PD, such as α-synuclein, is also relatively unexplored. While the mechanism(s) by which mutant LRRK2 causes neurodegeneration are not entirely certain, it is generally believed that disease-causing mutations may be associated with increased kinase activity, at least in situ in intact cells. However, a critical barrier to understanding the role of endogenous, wildtype LRRK2 in iPD is the absence of a practical, high-resolution assay for its activation state. We have developed and validated a pair of novel proximity ligation assays with excellent anatomical resolution that can rapidly provide information regarding activation state, cellular localization and physiological regulators of LRRK2. The assay is based on (i) S1292 phosphorylation and (ii) dissociation of 14-3-3 from LRRK2. Using this and other assays, we have preliminary evidence that (i) LRRK2 is activated in nigrostriatal neurons in iPD; (ii) sublethal concentrations of rotenone activate LRRK2; (iii) overexpression of α-synuclein in vivo activates LRRK2 in nigrostriatal neurons; (iv) LRRK2 activation is mediated by oxidative mechanisms. We now propose to examine the role of LRRK2 in iPD and its potential physiological interactions with α-synuclein and mitochondria. We will investigate the following questions: (1) What is the activation state of LRRK2 in human iPD brain tissue? (2) To what extent can endogenous LRRK2 activation be modeled in the rotenone rat? (3) Does!α-synuclein activate endogenous WT LRRK2? (4) Does oxidative stress activate LRRK2?

LRRK2的突变引起农场帕金森氏病（PD），在某些人群中，最多可能占全部的40％ 病例3。 LRRK2基因基因座还包含“特发性” PD（IPD）的危险因素；但是，lrrk2在 典型的IPD尚不清楚。此外，LRRK2与其他基因和与PD相关的蛋白质的关系，例如 α-突触核蛋白也相对出乎意料。而突变体LRK2引起神经退行性的机制 不确定，通常认为引起疾病的突变可能与激酶增加有关 活性，至少在完整的细胞中原位。但是，了解内源性，野生型的作用的关键障碍 IPD中的LRRK2是其激活状态的实际高分辨率测定法。 我们已经开发并验证了一对具有出色解剖分辨率的新型接近连接测定法 迅速提供了有关LRRK2的激活状态，细胞定位和物理调节剂的信息。 测定基于（I）S1292磷酸化和（II）从LRRK2解离14-3-3。使用这个和其他测定法，我们 有初步的证据表明，（i）LRRK2在IPD的黑质神经元中被激活； （ii）一定的浓度 紫藤酮激活LRRK2; （iii）体内α-突触核蛋白的过表达会激活黑质神经元中的LRRK2； （iv） LRRK2激活是由氧化机制介导的。现在，我们建议检查LRRK2在IPD及其ITS中的作用 潜在的生理相互作用与α-突触核蛋白和线粒体。我们将研究以下问题：（1） 人IPD脑组织中LRRK2的激活状态是什么？ （2）内源性LRRK2激活在多大程度上可以 在rotorone大鼠中建模？ （3）！α-突触核蛋白激活内源性WT LRRK2吗？ （4）氧化应激会激活 lrrk2？