LRRK2 and cellular pathways of Parkinson's Disease

LRRK2 与帕金森病的细胞通路

基本信息

批准号：
7268124
负责人：
WANLI W SMITH
金额：
$ 21.49万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-07-15 至 2008-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7268124
关键词：
Affect Biochemical Biological Assay Cell Death Signaling Process Cell Line Cell model Cells Chemicals Co-Immunoprecipitations Complex Cultured Cells Data Dominant Genes Environmental Risk Factor Gene Mutation Genetic Genetic Predisposition to Disease Intervention LRRK2 gene Label Lewy Bodies Mediating Methods Modification Molecular Molecular Target Mouse Cell Line Movement Disorders Mutation Nerve Degeneration Neurodegenerative Disorders Neurons Numbers PC12 Cells PINK1 gene Parkinson Disease Parkinson&apos s Disease Pathway Pathogenesis Pathology Pathway interactions Pattern Physiologic pulse Protein Kinase Interaction Proteins Pulse taking Recessive Parkinsonism, Autosomal Role Signal Pathway Signal Transduction Small Interfering RNA Study models Techniques Testing Therapeutic Intervention Toxic effect Transfection Ubiquitination aggregation pathway alpha synuclein dopaminergic neuron early onset experience human SNCAIP protein immunocytochemistry inhibitor/antagonist leucine-rich repeat kinase 2 mutant neurotoxicity parkin gene/protein protein aggregate protein aggregation protein degradation therapeutic target

项目摘要

DESCRIPTION (provided by applicant): Parkinson's disease (PD) is a progressive neurodegenerative movement disorder characterized by selective loss of dopaminergic neurons, and the presence of intracellular protein aggregates termed Lewy bodies. The pathogenesis of PD remains incompletely understood, but it appears to involve both genetic susceptibility and environmental factors. Five major genetic causes have been identified, including mutations in alpha-synuclein, parkin, PINK1, DJ-1 and LRRK2, and these mutations have provided models for study of the pathogenesis of genetic PD, with implications for sporadic PD. Mutations in LRRK2 (or dardarin) have been recently identified to cause autosomal dominant late onset, clinically typical PD with pleomorphic pathology. LRRK2 is a large complex protein, and contains domains that suggest roles for protein interactions, kinase activity, and Ras-related signaling. We have extensive experience in studying cell toxicity, signaling pathways, and protein interactions in neurodegenerative diseases. Our previous studies of PD have elucidated the role of alpha-synuclein toxicity, and protein interactions with synphilin-1 and parkin in the formation of Lewy body-like aggregates. We now propose to use similar methods to study LRRK2. We have evidence that LRRK2 interacts with other PD-related gene products and have promising data indicating that mutant LRRK2 can cause direct neuronal toxicity, providing a unique model for studying pathogenesis of LRRK2-related PD. In Specific Aim 1, we will study LRRK2 cellular expression and toxicity, and determine the signaling pathways by which LRRK2 induces neuronal degeneration. We will conduct cell transfection studies using wild type and mutant LRRK2, examine the LRRK2 distribution patterns, and confirm the LRRK2 toxicity. We will generate stable PC12 cell lines which inducibly express either wild type or mutant LRRK2 to study the molecular mechanisms underlying LRRK2 toxicity. In Specific Aim 2, we will test the functional effects of LRRK2 interaction with synphilin-1 and parkin on the pathways of cell toxicity and cellular aggregation using the cell models described in Aim 1. These studies will elucidate the functional role of LRRK2 in cell toxicity and the functional consequences of its interaction with synphilin-1 and parkin in cell culture, will help elucidate the pathogenesis of LRRK2-associated PD, and will identify potential therapeutic targets for intervention in PD.

描述（由申请人提供）：帕金森氏病（PD）是一种进行性神经退行性运动障碍，其特征是多巴胺能神经元的选择性丧失，并且存在被称为Lewy身体的细胞内蛋白质聚集体。 PD的发病机理仍未完全理解，但似乎涉及遗传易感性和环境因素。已经鉴定出五个主要的遗传原因，包括α-突触核蛋白，帕克蛋白，PINK1，DJ-1和LRRK2的突变，这些突变为研究遗传PD的发病机理提供了模型，对零星PD的影响。最近已经确定了LRRK2（或dardarin）中的突变，以引起常染色体显性晚期发作，临床上典型的具有多态性病理学的PD。 LRRK2是一种大型的复杂蛋白质，其中包含该结构域，这些结构域暗示了蛋白质相互作用，激酶活性和与RAS相关的信号传导的作用。我们在研究细胞毒性，信号通路和神经退行性疾病中的蛋白质相互作用方面具有丰富的经验。我们先前对PD的研究阐明了α-突触核蛋白毒性的作用，以及与Synphilin-1和Parkin在Lewy身体样骨料形成中的蛋白质相互作用。现在，我们建议使用类似的方法研究LRRK2。我们有证据表明LRRK2与其他与PD相关的基因产物相互作用，并具有有前途的数据表明突变体LRRK2可以引起直接的神经毒性，为研究LRRK2相关PD的发病机理提供了独特的模型。在特定目标1中，我们将研究LRRK2细胞表达和毒性，并确定LRRK2诱导神经元变性的信号传导途径。我们将使用野生型和突变体LRRK2进行细胞转染研究，检查LRRK2分布模式，并确认LRRK2毒性。我们将生成稳定的PC12细胞系，该细胞系可表达野生型或突变的LRRK2来研究LRRK2毒性的分子机制。在特定目标2中，我们将使用AIM 1中描述的细胞模型来测试LRRK2与Synphilin-1和Parkin相互作用与细胞毒性和细胞聚集途径的功能效应。这些研究将阐明LRK2在细胞毒性中的功能作用，以及其与Celliried and protiried partiried protiation-liluc and protiatient oss Assass-lycriatient oss Assass-aluc and sass Assass rucs inc sass Assass Incriast Inucs rucs and rucs and rucs inctiase的功能。 PD，并将确定潜在的治疗靶标在PD中进行干预。