Biology of Parkin and It's Role in Parkinson's Disease

帕金生物学及其在帕金森病中的作用

• 批准号: 8882845
• 负责人: Ted M. Dawson
• 金额: $ 41.31万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 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 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8882845
• 关键词: ABL1 gene; Adult; Affinity Chromatography; Alpha-Synuclein transgenic mouse; Amino Acids; Apoptosis; Biogenesis; Biology; Brain; Cells; Cessation of life; Complex; Data; Defect; Due Process; Event; Failure; Funding; Gene Proteins; Genes; Genetic; Genetic Transcription; Goals; Human; Impairment; Investigation; Knock-out; Knockout Mice; Lead; Mammals; Mediating; Methodology; Mitochondria; Modeling; Mus; Mutation; National Institute of Neurological Disorders and Stroke; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Nodal; Oxidative Stress; Parkin gene; Parkinson Disease; Pathogenesis; Patients; Phosphorylation; Play; Poly Adenosine Diphosphate Ribose; Polymerase; Process; Proteins; Proteomics; Quality Control; RNA; Recommendation; Research; Ribosomes; Role; Signal Transduction; Stable Isotope Labeling; Substantia nigra structure; TNFRSF5 gene; Technology; Therapeutic; Time; Toxic effect; Transcription Repressor/Corepressor; Transgenic Mice; Translating; Ubiquitin-Proteasomal Pathway; Ubiquitination; Up-Regulation; age related; alpha synuclein; base; biological adaptation to stress; c-abl Proto-Oncogenes; deep sequencing; disease-causing mutation; dopaminergic neuron; feeding; insight; loss of function; loss of function mutation; mitochondrial dysfunction; nitrosative stress; overexpression; oxidant stress; parkin gene/protein; prevent; small hairpin RNA; ubiquitin-protein ligase

PROJECT SUMMARY - PROJECT 1: BIOLOGY OF PARKIN AND ITS ROLE IN PARKINSON'S DISEASE Parkinson's disease (PD) is a complex neurodegenerative disorder that is both sporadic and familial. Mutations in parkin are the most common cause of autosomal recessive PD. In sporadic PD dopaminergic, oxidative and nitrosative stress as well as c-Abl phosphorylation result in inhibition of parkin. Thus, loss of parkin function is elemental to both familial and sporadic PD. Parkin is an E3 ligase, this loss of function leads to accumulation of the substrates, AIMP2 and PARIS. We have found that AIMP2 expression leads to age dependent DA neurodegeneration due to parthanatos. And PARIS expression may lead to loss of mitochondrial quality control that promotes neurodegeneration. Our hypothesis is that parkin inactivation in sporadic PD by nitrosative/oxidative stress, and c-Abl activation leads to phosphorylation of parkin on Y143 (pY143 parkin) and inactivation followed by the accumulation of parkin substrates, loss of mitochondrial quality control and toxicity. In parallel, α-synuclein is phosphorylated on Y39 (pY39 α-synuclein) resulting in aggregation and subsequent toxicity. Since aggregated α-synuclein can lead to mitochondrial dysfunction it creates a feed forward cycle. Aim 1: One of the unifying features of PD is mitochondrial dysfunction. This aim will explore the inter- relationship of PARIS and mitochondrial dysfunction caused by mutations in parkin. We have shown that PARIS is an important pathophysiologic substrate of parkin in PD that transcriptionally represses PGC-1α a major transcriptional co-activator that regulates mitochondrial biogenesis and mitochondrial oxidant stress responses. Aim 2: Inactivation of parkin results in accumulation of both AIMP2 and PARIS. Expression of either AIMP2 or PARIS is sufficient to promote age dependent DA neurodegeneration. The sequence of events activated by PARIS and AIMP2 will be explored to determine if and how these two proteins interact to initiate the cell death program, parthanatos. Aim 3: We observe pY143 parkin and elevated AIMP2 and PARIS in A53T α-synuclein transgenic mice that raises the question of whether parkin inactivation, PARIS and AIMP2 upregulation and PARP1 activation play a role in α-synuclein induced neurodegeneration? This possibility will be explored with the α-synuclein preformed fibrils (PFFs) model of PD. Aim 4: State-of-the-art technology including deep sequencing and SILAM (stable isotope labeling by amino acids in mammals) will be deployed to identify genes and proteins that are regulated by adult conditional knockout of parkin and their relationship to PARIS induction with the goal of identify nodal points in the signal cascade of neurodegeneration that can provide new targets for the treatment of PD.

项目摘要 - 项目 1：帕金森生物学及其在帕金森病中的作用 帕金森病 (PD) 是一种复杂的神经退行性疾病，具有散发性和家族性。 Parkin 突变是常染色体隐性遗传 PD 的最常见原因。 氧化和亚硝化应激以及 c-Abl 磷酸化会导致 Parkin 的抑制。 Parkin 功能对于家族性和散发性 PD 至关重要 Parkin 是一种 E3 连接酶，这种功能丧失会导致。 底物 AIMP2 和 PARIS 的积累我们发现 AIMP2 表达会导致年龄。 PARIS 表达可能导致 DA 神经变性的丧失。 促进神经变性的线粒体质量控制。 我们的假设是，散发性 PD 中的 Parkin 失活是由于亚硝化/氧化应激和 c-Abl 激活所致 导致 Y143 (pY143 Parkin) 上的 Parkin 磷酸化并失活，随后积累 Parkin 底物、线粒体质量控制丧失和毒性同时，α-突触核蛋白被磷酸化。 由于聚集的 α-突触核蛋白，Y39（pY39 α-突触核蛋白）会导致聚集和随后的毒性。 可能导致线粒体功能障碍，它会产生前馈循环。 目标 1：PD 的共同特征之一是线粒体功能障碍。该目标将探讨线粒体功能障碍之间的相互作用。 PARIS 与 Parkin 突变引起的线粒体功能障碍的关系。 PARIS 是 PD 中 Parkin 的重要病理生理底物，可转录抑制 PGC-1α 调节线粒体生物发生和线粒体氧化应激的主要转录辅激活因子 回应。 目标 2：parkin 失活导致 AIMP2 和 PARIS 表达累积。 或 PARIS 足以促进年龄依赖性 DA 神经变性激活的事件序列。 PARIS 和 AIMP2 将被探索以确定这两种蛋白质是否以及如何相互作用来启动细胞 死亡计划，帕塔纳托斯。 目标 3：我们在 A53T α-突触核蛋白转基因小鼠中观察到 pY143 Parkin 以及 AIMP2 和 PARIS 升高， 提出了 Parkin 失活、PARIS 和 AIMP2 上调以及 PARP1 激活是否发挥作用的问题 在α-突触核蛋白诱导的神经变性中的作用？将用α-突触核蛋白探索这种可能性？ PD 的预制原纤维 (PFF) 模型。 目标 4：最先进的技术，包括深度测序和 SILAM（氨基稳定同位素标记） 哺乳动物中的酸）将被用来识别受成人条件调节的基因和蛋白质 脱离 Parkin 及其与 PARIS 感应的关系，目的是识别信号中的节点敲击点 神经退行性变的级联反应可以为帕金森病的治疗提供新的靶点。