High Throughput Screening to Discover Chemical Probes and Pharmacological Agents for Modulating Parkin Activity

高通量筛选以发现调节 Parkin 活性的化学探针和药理学制剂

• 批准号: 9115654
• 负责人: XUEDONG LIU
• 金额: $ 29.93万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9115654
• 关键词: Accounting; Apoptotic; Biochemical; Biological; Biological Assay; Biological Models; Cells; Chemicals; DNA Sequence Alteration; Development; Disease; Environmental Risk Factor; Genes; Genetic; Goals; Health; In Vitro; Juvenile Parkinson Disease; LRRK2 gene; Lead; Libraries; Mitochondria; Molecular; Molecular Target; Motor Neurons; Mutation; Neurodegenerative Disorders; PINK1 gene; Paraquat; Parkinson Disease; Pathway interactions; Phosphotransferases; Polyubiquitination; RNA Interference; Reagent; Research; Risk; Role; Specificity; System; Validation; Valinomycin; alpha synuclein; analog; base; chemical genetics; early onset; genetic approach; high throughput screening; inhibitor/antagonist; mutant; novel; novel therapeutic intervention; novel therapeutics; parkin gene/protein; parkin protein; prevent; protein aggregate; protein aggregation; protein misfolding; protein protein interaction; reconstitution; response; screening; small molecule; small molecule inhibitor; tool; ubiquitin-protein ligase

 DESCRIPTION (provided by applicant): Many neurodegenerative diseases are known to be associated with inappropriate protein aggregation. In Parkinson's disease (PD), formation of protein aggregates in motor neurons is a hallmark of the disease. Both genetic and environmental factors have been associated with an increased risk of PD. Several susceptible genes, including alpha-Synuclein, Parkin, PINK1, LRRK2, DJ in familial PD have been identified. Mutations in Parkin are responsible for an early- onset autosomal recessive form (autosomal recessive juvenile parkinsonism; AR-JP) of PD and account for 50% all recessively transmitted early-onset PD cases. Parkin protein is a ubiquitin E3 ligase and has been shown to catalyze polyubiquitination of a variety of substrates. Parkin appears to be an integral component of cellular defense systems against misfolded proteins and damaged mitochondria. Perturbation of the Parkin activity by environmental factors or genetic mutations is the compelling mechanism for a subset of Parkinson's disease. We hypothesized that systematic identification of small molecule probes that modulate the molecular pathways that control Parkin mitochondrial localization will help to unravel the type of environmental factors that may be associated with the development of Parkinson's disease. In addition, availability of such specific tool compounds can help to clarify the roles of Parkin mitochondrial recruitment and mitophagy in the onset and progression of PD in model systems. We further hypothesized that identification of small-molecule inhibitors that can prevent mutant Parkin aggregation may yield nove chemical probes for the pathways that regulates Parkin aggregation and may offer new therapeutic strategies for Parkinson's disease. Currently there are no specific pharmacological agents that can block wild type Parkin recruitment to mitochondria upon depolarization, neither are specific inhibitors that can block aggregation of Parkin mutants in cells. The overall goal of this application is to develop a set of chemical probes to permit development of the chemical genetics approaches to dissect the function of PINK1 and Parkin in Parkinson's disease. Specificity and counter screening assays will be implemented to eliminate non-specific compounds. Deconvolution assays will be developed to allow determination of the molecular targets of the chemical probes discovered in related HTS cell based assays.

 描述（由申请人提供）：已知许多神经退行性疾病与不适当的蛋白质聚集有关，在帕金森病（PD）中，运动神经元中蛋白质聚集的形成是该疾病的标志，遗传因素和环境因素都与该疾病有关。一些易感基因，包括家族性 PD 中的 α-突触核蛋白、Parkin、PINK1、LRRK2、DJ，已被确定为导致 PD 风险增加的原因。早发性常染色体隐性遗传型帕金森病（常染色体隐性幼年型帕金森病；AR-JP），占所有隐性传播的早发性帕金森病病例的 50%。 Parkin 蛋白是一种泛素 E3 连接酶，已被证明可催化多种多泛素化。 Parkin 似乎是细胞防御系统的一个组成部分，可抵抗环境因素或遗传对错误折叠蛋白质和受损线粒体的干扰。突变是帕金森病的一个重要机制，我们致力于系统地鉴定调节控制帕金线粒体定位的分子途径的小分子探针，这将有助于揭示可能与帕金森病发展相关的环境因素类型。此外，此类特定工具化合物的可用性有助于阐明 Parkin 线粒体募集和线粒体自噬在模型系统中 PD 发生和进展中的作用，我们进一步鉴定了可以预防突变的小分子抑制剂。 Parkin 聚集可能会产生新的化学探针，用于调节 Parkin 聚集的途径，并可能为帕金森病提供新的治疗策略。目前，没有特定的药物可以阻止野生型 Parkin 在去极化时招募到线粒体，也没有可以阻止的特定抑制剂。该应用的总体目标是开发一套化学探针，以允许开发化学遗传学方法来剖析 PINK1 和 PINK1 的功能。帕金森氏病中的帕金氏病将进行特异性和反筛选测定，以消除非特异性化合物，从而确定相关 HTS 细胞测定中发现的化学探针的分子靶标。