Regulation of LRRK2 in lysosomal stress response

LRRK2 在溶酶体应激反应中的调节

• 批准号: 10592134
• 负责人: TSO-PANG YAO
• 金额: $ 24.15万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10592134
• 关键词: Alleles; Biochemical; Cell model; Cells; Chemicals; Chloroquine; Complex; Disease; Drosophila genus; Elements; Endosomes; Event; Feedback; Genetic; Golgi Apparatus; Homeostasis; Idiopathic Parkinson Disease; Impairment; LRRK2 gene; Lesion; Lipids; Lysosomes; Mediating; Membrane; Metabolism; Mitochondria; Modeling; Molecular; Mutation; Nerve Degeneration; Parkinson Disease; Pathogenesis; Pathogenicity; Penetrance; Phosphorylation; Phosphotransferases; Process; Production; Proteins; Regulation; Resolution; Role; Signal Pathway; Signal Transduction; Sphingolipids; Sphingomyelins; Stress; Toxic effect; biological adaptation to stress; inhibitor; mutant; novel; phosphatidylinositol 4-phosphate; protein kinase D; recruit; response; stressor; trafficking

The pathophysiological basis of selective neurodegeneration in Parkinson's disease remains controversial. Recent genetic and biochemical analyses have uncovered aberrant LRRK2 kinase activity as a salient feature of late-onset Parkinson's disease (PD) of both familial and idiopathic origin. Familial LRRK2 mutations are generally associated with elevated kinase activity. Identifying the pathophysiological processes that regulate LRRK2 kinase could provide a critical window to understand the pathogenesis unique to PD. LRRK2 is recruited and activated at enlarged lysosomes induced by lysosome stressors, chloroquine (CO) and LLOMe. We hypothesize that LRRK2 recruitment to the stressed lysosome is part of the lysosome stress response that acts to return lysosome to the basal states. LRRK2 association with stressed lysosomes is typically transient Remarkably, the LRRK2 association with stressed lysosomes becomes dramatically enhanced in PD-associated Vps35-D620N mutant cells. This finding suggests that LRRK2 becomes excessively activated by the lysosomal stress in Vps35-D620N cells. Thus, stress-induced LRRK2-lysosome association and activation are tightly regulated, and this regulation is disrupted by a mutation that causes familial PD. A small focused chemical inhibitor screen for additional regulators of LRRK2-lysosome association identified PKD. PKD, similar to LRRK2, is activated by lysosomal stresses, while its inactivation led to aberrant LRRK2-lysosome association and activation. This proposal aims to identify the molecular entity that recruits LRRK2 to stressed lysosomes and characterize the role of PKD-signaling pathway in LRRK2 activation and pathogenic toxicity. This proposal's completion could provide a biochemical mechanism underlying pathogenic LRRK2 activation, identify PKD as a novel regulator of LRRK2, and ascertain lysosomal stress as a pathophysiological trigger to late-onset PD

帕金森氏病中选择性神经退行性的病理生理基础仍然存在争议。最近的遗传和生化分析发现了异常的LRRK2激酶活性，这是家族性和特发性起源后期发作的帕金森氏病（PD）的显着特征。家族性LRRK2突变通常与激酶活性升高有关。确定调节LRRK2激酶的病理生理过程可以提供 了解PD独有的发病机理的关键窗口。 LRRK2在由溶酶体胁迫，氯喹（CO）和Llome诱导的增大溶酶体上募集和激活。我们假设LRRK2募集到应力的溶酶体是溶酶体应激反应的一部分，该溶酶体应激反应可将溶酶体返回到基础状态。 LRRK2与应激溶酶体的关联通常是瞬时的，在PD相关的VPS35-D620N突变细胞中，LRRK2与应激溶酶体的关联显着增强。这一发现表明，LRRK2在VPS35-D620N细胞中的溶酶体应激过度激活。因此，应力诱导的LRRK2溶酶体 关联和激活受到严格调节，该调节受到导致家族性PD的突变的破坏。一个小的聚焦化学抑制剂筛选，用于鉴定出PKD的LRRK2溶酶体关联的其他调节剂。与LRRK2相似的PKD被溶酶体应激激活，而其失活导致异常LRRK2溶质体的关联和激活。该提案旨在确定募集LRRK2强调溶酶体的分子实体，并表征PKD信号途径在LRRK2激活和致病性毒性中的作用。该提案的完成可以提供致病性LRRK2激活的生化机制，将PKD识别为LRRK2的新调节剂，并确定溶酶体应激作为病理生理触发的溶酶体应激触发剂量延迟触发PD