The PARK10 gene USP24 affects Parkinson's Disease via regulation of autophagy

PARK10 基因 USP24 通过调节自噬影响帕金森病

• 批准号: 8931076
• 负责人: MARTA M LIPINSKI
• 金额: $ 7.68万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-30 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8931076
• 关键词: 1-Phosphatidylinositol 3-Kinase; Affect; Attenuated; Autophagocytosis; Biology; Cell Death; Cell Line; Cell Survival; Cell model; Cell physiology; Cells; Code; Data; Defect; Degradation Pathway; Development; Disease model; Gene Targeting; Genes; Genetic Polymorphism; Genomics; Goals; Health; Herbicides; Human; Human Cell Line; Human Genome; Immunofluorescence Immunologic; In Vitro; Intervention; Libraries; Link; Lysosomes; Mitochondria; Modeling; Molecular; Neurodegenerative Disorders; Neurons; Outcome; PARK10 gene; Paper; Paraquat; Parkinson Disease; Pathology; Pathway interactions; Patients; Peptide Hydrolases; Pharmaceutical Preparations; Play; Predisposition; Prevalence; Prevention; Prevention strategy; Proteins; Rattus; Reagent; Regulation; Research; Single Nucleotide Polymorphism; Small Interfering RNA; Substantia nigra structure; Testing; Time; Ubiquitin; Up-Regulation; Western Blotting; age related; alpha synuclein; base; chemical genetics; citizen science; effective therapy; genetic manipulation; genome-wide; inhibition of autophagy; knock-down; mutant; novel; overexpression; prevent; protein degradation; risk variant; synuclein; treatment strategy; vector

DESCRIPTION (provided by applicant): Recent studies indicate a causative link between defects in autophagy, a lysosome-dependent intracellular degradation pathway, and Parkinson's disease (PD). Conversely, up-regulation of autophagy has been proposed as a possible PD intervention strategy. We recently identified USP24, a gene of unknown function located in the PD associated PARK10 locus, as a negative regulator of autophagy. Our data indicate that USP24 protein levels are increased in the substantia nigra of a subpopulation of PD patients, suggesting possible involvement also in idiopathic PD. On the other hand, inactivation of USP24 induces autophagy and can play a protective function in cellular models of PD. The goal of our proposal is to use in vitro PD models to test the hypothesis that USP24 affects PD predisposition via autophagy-dependent mechanism and that inhibition of USP24 can attenuate PD pathology. AIM 1 will determine how USP24 overexpression and PD-associated coding region polymorphisms alter levels of autophagy. We hypothesize that USP24 inhibits autophagy flux by interfering with function of the type III PI3 kinase. We will test this by expressing wild-type and mutant USP24 in human cell lines and primary rat neurons and using immunofluorescence, western blot and autophagic flux analysis to investigate the effects on the levels of autophagy, type III PI3 kinase activity, lysosomal degradation and mitochondrial function. AIM 2 will determine the influence of USP24-regulated autophagy on outcomes in cell based PD models. We hypothesize that induction of autophagy by USP24 inhibition will attenuate PD pathology. To test this we will evaluate the effects of USP24 knock-down, overexpression and PD polymorphisms on cell viability, mitochondrial function and protein turnover in two cell based PD models: treatment with PD associated herbicide paraquat and expression of mutant alpha-synuclein A53T. Our data will be the necessary first step to understand how USP24 regulates autophagy and contributes to PD and to determine if up-regulation of autophagy by inhibition of USP24 can be used as a novel prevention and treatment strategy.

描述（由申请人提供）：最近的研究表明自噬缺陷、溶酶体依赖性细胞内降解途径与帕金森病 (PD) 之间存在因果关系。相反，自噬的上调已被提议作为一种可能的 PD 干预策略。我们最近发现 USP24（一种功能未知的基因，位于 PD 相关 PARK10 位点）是自噬的负调节因子。我们的数据表明，PD 患者亚群的黑质中 USP24 蛋白水平升高，表明也可能参与特发性 PD。另一方面，USP24 失活可诱导自噬，并可在 PD 细胞模型中发挥保护功能。我们提案的目标是使用体外 PD 模型来检验 USP24 通过自噬依赖性机制影响 PD 易感性以及抑制 USP24 可以减弱 PD 病理学的假设。 AIM 1 将确定 USP24 过表达和 PD 相关编码区多态性如何改变自噬水平。我们假设 USP24 通过干扰 III 型 PI3 激酶的功能来抑制自噬通量。我们将通过在人类细胞系和原代大鼠神经元中表达野生型和突变型 USP24 并使用免疫荧光、蛋白质印迹和自噬流分析来研究其对自噬、III 型 PI3 激酶活性、溶酶体降解和线粒体功能水平的影响来测试这一点。 AIM 2 将确定 USP24 调节的自噬对基于细胞的 PD 模型结果的影响。我们假设通过抑制 USP24 诱导自噬将减弱 PD 病理。为了测试这一点，我们将评估 USP24 敲低、过度表达和 PD 多态性对两种基于细胞的 PD 模型中细胞活力、线粒体功能和蛋白质更新的影响：用 PD 相关除草剂百草枯处理和突变 α-突触核蛋白 A53T 的表达。我们的数据将是了解 USP24 如何调节自噬并促进 PD 的必要的第一步，并确定是否可以将通过抑制 USP24 上调自噬作为一种新的预防和治疗策略。