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：
8071041
负责人：
Salvatore James Cherra
金额：
$ 1.25万
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 2011-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8071041
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 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 Synaptic plasticity 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 neuroblastoma cell neuroprotection overexpression pre-doctoral prevent protective effect protein profiling reuptake synaptic function therapeutic target vector

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 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 Synaptic plasticity 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 neuroblastoma cell neuroprotection overexpression pre-doctoral prevent protective effect protein profiling reuptake synaptic function therapeutic target vector

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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）：神经退行性疾病使数百万个人从儿童到老年人。没有已知的原因或有效的疗法，这种类别的疾病超过了大脑的补偿性神经元可塑性的能力，其结果令人衰弱。与神经退行性相关的两种途径是线粒体功能障碍和大型噬菌体。这项研究建议着重于这两种途径作为轴突和树突变性的调节剂（神经突变性）之间的相互作用。宏观哲学（以下称为自噬）是包括线粒体在内的长寿命蛋白和细胞器的大量分解代谢。尽管健康需要基础自噬水平，但失调已被视为导致神经突变性的原因。线粒体是无处不在的细胞器，在神经元的适当功能和可塑性中起关键作用。大型噬菌体和线粒体功能障碍/损失的失调与帕金森氏病（PD）的发病机理有关。最近的研究说明了PTEN诱导的激酶1（PINK1）的神经保护活性。此外，PINK1与常染色体隐性PD有关，大概是由于功能突变的丧失而链接。我假设PINK1信号传导通过其对自噬/线粒体的影响来调节神经突变性。通过生化和显微镜研究，将在体外测量PINK1在调节神经突变性，自噬和线粒体损失中的作用。 AIM 1将确定PINK1表达水平是否调节自噬并预防导致神经突变性的神经元损伤。 AIM 2将确定PINK1敲低或与疾病相关的突变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进行神经突变性。 AIM 3将使用非偏置蛋白质组方法来识别PINK1的下游线粒体靶标。将分别比较粉红色的小鼠或细胞和野生型小鼠或细胞之间的磷酸化蛋白质谱。潜在的下游PINK1靶标将在野生型样品中鉴定为pink1缺乏样品中降低的磷蛋白。使用RNAi，我将确定PINK1神经保护作用的推定下游PINK1介体是否需要。通过确定调节PINK1介导的神经保护和自噬的“自我消化”的潜在途径，这项研究的结果将洞悉帕金森氏症和相关疾病中神经突变性的机制。除了向申请人提供专业培训外，该研究建议还旨在发现有希望的药物靶标以治疗神经退行性疾病。