Elucidating Autophagy-lysosome Mechanism in LRRK2 and alpha-synuclein Pathogenic Pathways

阐明 LRRK2 和 α-突触核蛋白致病途径中的自噬溶酶体机制

• 批准号: 9017208
• 负责人: Zhenyu Yue
• 金额: $ 36.9万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9017208
• 关键词: 1-Phosphatidylinositol 3-Kinase; Affect; Animal Model; Autophagocytosis; Autophagosome; Axon; Brain; Cell model; Collaborations; Complex; Data; Defect; Dopamine; Endosomes; Excision; Exocytosis; Gene Expression; Genes; Genetic; Genetic screening method; Goals; Guanosine Triphosphate Phosphohydrolases; Homeostasis; Human; In Vitro; Kidney; Knockout Mice; Knowledge; LRRK2 gene; Link; Lysosomes; Mediating; Membrane Protein Traffic; Molecular; Molecular Genetics; Mus; Mutation; Neurites; Neurons; Organelles; Outcome; Parkinson Disease; Pathogenesis; Pathway interactions; Phenocopy; Phosphatidylinositols; Phosphotransferases; Process; Protein-Serine-Threonine Kinases; Proteins; Recycling; Regulation; Reporting; Research; Rodent; Rodent Model; Role; SNAP receptor; Structure; Testing; Therapeutic; Transgenic Mice; Transgenic Organisms; abstracting; age related; aged; alpha synuclein; base; biochemical model; experience; in vivo; injured; interdisciplinary approach; mouse model; mutant; neuroprotection; neurotoxicity; overexpression; parkin gene/protein; protein aggregate; protein transport; trafficking; transcription factor; transmission process

Project Summary/Abstract Our goal is to elucidate autophagy-lysosomal mechanism in the pathogenesis of Parkinson's disease (PD). Early pathological analysis of PD and recent studies of PD-linked genes, such as SNCA, LRRK2, Parkin, Pink1, GBA and ATP13A2, implicates dysfunctional autophagy-lysosomal pathway in the pathogenesis of PD. Recent evidence shows that inhibition of LRRK2 or expression of LRRK2 PD mutants causes aberrant autophagic activity, but the detailed mechanism is unclear. Our preliminary study suggests a link of LRRK2 to an autophagy kinase, which is essential for autophagy and required for axon/neurite outgrowth. Here we propose to investigate the molecular mechanism whereby LRRK2 regulates autophagy through directly targeting autophagy machinery and dysfunctional autophagy as a potential PD pathogenic pathway. We previously reported that disruption of autophagy in mice leads to age-dependent accumulation of endogenous α-Syn in dystrophic axons and altered dopamine transmission, consist with impaired autophagy as one of the failing cellular mechanisms involved in the pathogenesis of PD. Emerging evidence suggests that autophagy not only participates in degradation of α-Syn, but also is involved in the secretion pathway of α-Syn, which underlies the spreading of synucleiopathy. Fibrillar α-Syn can be secreted through exosomes and/or autophagosome-related structures. Furthermore, increasing autophagy gene beclin1 or transcription factor TFEB expression facilitates the autophagic clearance of α-Syn and offers neuroprotection in rodent models. However, the precise mechanism for the autophagic clearance of intracellular α-Syn remains poorly characterized. Finally, whether LRRK2 also contributes to the control of α-Syn homeostasis through regulation of autophagy is unclear. Given the emerging role of autophagy in the exocytosis of α-Syn, we hypothesize that autophagy regulates α-Syn clearance through both degradative and secretory pathways; LRRK2 modulates α- Syn homeostasis by targeting autophagy. In Project 3 we propose (1) to determine molecular and genetic basis for LRRK2-mediated autophagy regulation. We will use biochemical, cellular and animal models to dissect the role for LRRK2 wildtype and PD mutation G2019S in autophagy control through modulating the autophagy kinase and SNARE-related protein trafficking; (2) to determine the mechanism that autophagy and LRRK2 modulate α-Syn homeostasis by targeting multiple trafficking pathways. We will apply multidisciplinary approaches to test that LRRK2 and autophagy pathway converge to regulate α-Syn homeostasis by targeting secretion and degradation in cell and animal models of PD. Our project 3 will be conducted in close collaboration with Drs. Jie Shen and Tom Südhof, who have extensive experience in LRRK2 and α-Syn research, respectively. Completion of our project 3 is expected to provide timely knowledge for understanding LRRK2 pathogenic mechanism and developing potential therapeutic strategies by targeting autophagy-lysosome pathways for the clearance of α-synuclein.

项目摘要/摘要 我们的目标是阐明帕金森氏病（PD）病原体中的自噬 - 溶酶体机制。 PD的早期病理分析和PD连锁基因的最新研究，例如SNCA，LRRK2，Parkin， PINK1，GBA和ATP13A2暗示了PD发病机理中功能失调的自噬溶质体途径。 最近的证据表明，抑制LRRK2或表达LRRK2 PD突变体会导致异常 自噬活动，但详细的机制尚不清楚。 一种自噬激酶，对于自噬至关重要，对于轴突/神经突生长所必需。 建议研究LRRK2定期自噬直接thursy的分子机制 靶向自噬机制和功能障碍自噬作为潜在的致病途径 据详细报道，小鼠自噬的纪律导致内源性的年龄依赖性积累 α-Syn在营养不良轴突和多巴胺传播中，由弱势自噬组成 涉及PD发病机理的细胞机制表明自噬。 不仅参与α-syn的降解，而且还参与了α-syn的秘密途径， 是突触病的传播的基础。 自噬相关结构。 TFEB表达促进了α-Syn的自噬清除率，并在啮齿动物模型中提供神经保护作用。 但是，细胞内α-Syn自噬清除的确切机制仍然很差 最终。 鉴于新兴的作用，自噬尚不清楚。 自噬通过降解和分泌途径调节α-syn清除率。 通过靶向自噬，在项目3中，我们提出（1）来确定分子和遗传 LRK2介导的自噬调节的基础。 通过调节您，剖析了LRRK2野生型和突变G2019在自噬控制中的作用 自噬激酶和与圈圈相关的蛋白质运输； LRRK2模块化α-Syn稳态通过靶向多个运输途径 多学科测试LRRK2和自噬途径的多学科方法会收敛以调节α-Syn 通过靶向PD的细胞和动物模型中的分泌和降解。 与博士密切合作。 LRRK2和α-Syn研究，尊重者。 用于了解LRRK2致病机制并通过靶向发展潜在的治疗策略 自噬 - 溶质体途径可清除α-突触核蛋白。