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

基本信息

批准号：
8822475
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
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-30 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8822475
关键词：
1-Phosphatidylinositol 3-Kinase Affect Attenuated Autophagocytosis Biology Cell Death Cell Line Cell Survival Cell model Cell physiology Cells Code Communities Data Defect Degradation Pathway Development Disease model Gene Targeting Genes Genetic Polymorphism Genomics Goals 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 Science Single Nucleotide Polymorphism Small Interfering RNA Substantia nigra structure Testing Time Ubiquitin Up-Regulation Western Blotting age related alpha synuclein base chemical genetics effective therapy genetic manipulation genome-wide inhibition of autophagy knock-down mutant novel overexpression prevent protein degradation public health relevance risk variant synuclein treatment strategy vector

项目摘要

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氏病（PD）之间存在原因联系。相反，已经提出了自噬的上调作为可能的PD干预策略。我们最近确定了USP24，这是一个位于PD相关PARK10基因座中的未知功能的基因，是自噬的负调节剂。我们的数据表明，在PD患者亚群的基本中，USP24蛋白水平升高，这表明可能参与特发性PD。另一方面，USP24的失活会诱导自噬，并且可以在PD的细胞模型中发挥保护作用。我们建议的目的是使用体外PD模型来测试USP24通过自噬依赖性机制影响PD易感性的假设，并且对USP24的抑制可以减弱PD病理。 AIM 1将确定USP24过表达和与PD相关的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发性大鼠神经元中表达野生型和突变体USP24，并使用免疫荧光，蛋白质印迹和自噬通量分析来研究对自噬，III型PI3激酶活性水平的影响，溶酶体脱落和线粒体功能。 AIM 2将确定USP24调节的自噬对基于细胞PD模型的结果的影响。我们假设USP24抑制对自噬的诱导会减弱PD病理。为了测试这一点，我们将评估USP24敲低，过表达和PD多态性对两个基于细胞的PD模型中细胞活力，线粒体功能和蛋白质的影响：使用PD相关的除草剂paraquat和突变体α-突变型核蛋白A53T的表达。我们的数据将是了解USP24如何调节自噬并有助于PD的必要第一步，并确定是否可以通过抑制USP24来上调自噬的上调作为新颖的预防和治疗策略。