PINK1 in the Regulation of Macroautophagy and Parkinsonian Neurodegeneration.

PINK1 在巨自噬和帕金森神经变性的调节中的作用。

基本信息

批准号：
7614733
负责人：
Salvatore James Cherra
金额：
$ 4.17万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-05-01 至 2012-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7614733
关键词：
1-Methyl-4-phenylpyridinium Abbreviations Affect Alzheimer&apos s Disease Apoptotic Autophagocytosis Axon Biochemical Brain Catabolism Cell Line Cells Child Clinical Cultured Cells Data Dendrites Disease Dopamine Drug Delivery Systems Elderly Functional disorder Gene Mutation Genes Green Fluorescent Proteins Health High temperature of physical object Huntington Disease Immunofluorescence Microscopy Impairment In Vitro Individual Induced Mutation Injury Knockout Mice LRRK2 gene Lead Length Life Light Link Maintenance Mass Spectrum Analysis Measures Mediating Mediator of activation protein Microscopy Microtubule-Associated Proteins Mitochondria Morphology Mus Mutation Nerve Degeneration Neurites Neuroblastoma Neurodegenerative Disorders Neuronal Injury Neuronal Plasticity Neurons Neurotoxins Organelles Oxidopamine PINK1 gene PTEN gene Parkinson Disease Parkinsonian Disorders Pathogenesis Pathway interactions Patients Phosphoproteins Phosphotransferases Play Proteins RNA Interference RNAi vector Regulation Relative (related person)Research Proposals Role Sampling Signal Pathway Signal Transduction Synapses Toxic effect Toxin Training Wild Type Mouse design effective therapy inhibitor/antagonist insight leucine-rich repeat kinase 2 loss of function mutation mitochondrial autophagy mitochondrial dysfunction mutant neuroprotection overexpression pre-doctoral prevent protective effect protein profiling reuptake synaptic function therapeutic target vector

项目摘要

DESCRIPTION (provided by applicant): Neurodegenerative diseases afflict millions of individuals ranging from children to the elderly. Without known causes or effective therapies, this class of diseases outstrips the brain's capacity for compensatory neuronal plasticity with debilitating results. Two pathways involved with neurodegeneration are mitochondrial dysfunction and macroautophagy. This research proposal focuses on the interplay between these two pathways as regulators of axonal and dendritic degeneration (neurite degeneration). Macroautophagy (hereafter, autophagy) is the bulk catabolism of long-lived proteins and organelles, including mitochondria. While basal levels of autophagy are required for health, dysregulation has been implicated as a cause for neurite degeneration. Mitochondria are ubiquitous organelles that play key roles in the proper function and plasticity of neurons. Dysregulated macroautophagy and mitochondrial dysfunction/loss have been implicated in the pathogenesis of Parkinson's disease (PD). Recent studies have illustrated the neuroprotective activity of the PTEN-induced kinase 1 (PINK1). Additionally, PINK1 has been linked to autosomal recessive PD, presumably through loss of function mutations. I hypothesize that PINK1 signaling regulates neurite degeneration through its effects on autophagy/mitophagy. The role of PINK1 in regulating neurite degeneration, autophagy, and mitochondrial loss will be measured in vitro by biochemical and microscopy studies. Aim 1 will determine whether levels of PINK1 expression regulate autophagy and protect against neuronal injuries that cause neurite degeneration. Aim 2 will determine whether PINK1 knockdown or disease-associated mutations induce autophagy and neurite degeneration. I will also determine whether PINK1 knockdown or mutants require autophagy for neurite degeneration. Aim 3 will use a non-biased proteomies approach to identify downstream mitochondrial targets of PINK1. Phosphorylated protein profiles will be compared between PINK1 deficient mice or cells and wild type mice or cells, respectively. Potential downstream PINK1 targets will be identified as phosphoproteins in wild-type samples that are reduced in PINK1 deficient samples. Using RNAi, I will determine if the putative downstream PINK1 mediators are required for the PINK1 neuroprotective effects. By identifying potential pathways that regulate PINK1-mediated neuroprotection and autophagic "selfdigestion", the results of this study will provide insight into the mechanisms of neurite degeneration in Parkinson's and related diseases. In addition to providing predoctoral training to the applicant, this research proposal is designed to uncover promising drug targets for the treatment of neurodegenerative diseases.

描述（由申请人提供）：神经退行性疾病困扰着从儿童到老年人的数百万人。如果没有已知的原因或有效的治疗方法，这类疾病会超出大脑补偿神经元可塑性的能力，从而导致衰弱的结果。与神经变性有关的两条途径是线粒体功能障碍和巨自噬。该研究计划重点关注这两种途径作为轴突变性和树突变性（神经突变性）调节剂之间的相互作用。巨自噬（以下简称自噬）是长寿命蛋白质和细胞器（包括线粒体）的大量分解代谢。虽然自噬的基础水平是健康所必需的，但失调已被认为是神经突退化的原因。线粒体是普遍存在的细胞器，在神经元的正常功能和可塑性中发挥着关键作用。巨自噬失调和线粒体功能障碍/丢失与帕金森病 (PD) 的发病机制有关。最近的研究表明 PTEN 诱导的激酶 1 (PINK1) 具有神经保护活性。此外，PINK1 可能通过功能缺失突变与常染色体隐性遗传性 PD 相关。我假设 PINK1 信号通过其对自噬/线粒体自噬的影响来调节神经突变性。 PINK1 在调节神经突变性、自噬和线粒体丢失中的作用将通过生化和显微镜研究在体外测量。目标 1 将确定 PINK1 表达水平是否调节自噬并防止导致神经突变性的神经元损伤。目标 2 将确定 PINK1 敲低或疾病相关突变是否会诱导自噬和神经突变性。我还将确定 PINK1 敲低或突变体是否需要自噬来导致神经突退化。目标 3 将使用无偏蛋白质组学方法来识别 PINK1 的下游线粒体靶标。将分别比较 PINK1 缺陷小鼠或细胞与野生型小鼠或细胞之间的磷酸化蛋白质谱。潜在的下游 PINK1 靶标将被鉴定为野生型样品中的磷蛋白，这些磷蛋白在 PINK1 缺陷样品中会减少。使用 RNAi，我将确定 PINK1 神经保护作用是否需要假定的下游 PINK1 介质。通过确定调节 PINK1 介导的神经保护和自噬“自我消化”的潜在途径，这项研究的结果将深入了解帕金森病及相关疾病的神经突变性机制。除了为申请人提供博士前培训外，该研究计划还旨在发现治疗神经退行性疾病的有希望的药物靶点。