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

帕金森病选择性神经退行性变的病理生理学基础仍然存在争议。最近的遗传和生化分析发现，异常LRRK 2激酶活性作为一个显着的特点，迟发性帕金森氏病（PD）的家族性和特发性起源。家族性LRRK 2突变通常与激酶活性升高相关。确定调节LRRK 2激酶的病理生理过程可以提供 了解PD独特发病机制的关键窗口。LRRK 2在由溶酶体应激物氯喹（CO）和LLOMe诱导的扩大的溶酶体处被募集和活化。我们假设LRRK 2向应激溶酶体的募集是溶酶体应激反应的一部分，其作用是使溶酶体返回到基础状态。LRRK 2与应激溶酶体的结合通常是短暂的。值得注意的是，LRRK 2与应激溶酶体的结合在PD相关的Vps 35-D 620 N突变细胞中显著增强。这一发现表明，LRRK 2在Vps 35-D 620 N细胞中被溶酶体应激过度激活。因此，应激诱导的LRRK 2-溶酶体 关联和激活受到严格调节，并且这种调节被导致家族性PD的突变破坏。一个小的集中化学抑制剂筛选LRRK 2-溶酶体协会的其他监管机构确定PKD。PKD类似于LRRK 2，被溶酶体应激激活，而其失活导致异常的LRRK 2-溶酶体缔合和激活。该提案旨在鉴定将LRRK 2募集到应激溶酶体的分子实体，并表征PKD信号通路在LRRK 2活化和致病毒性中的作用。该提案的完成可以提供致病性LRRK 2激活的生化机制，鉴定PKD作为LRRK 2的新调节剂，并确定溶酶体应激作为迟发性PD的病理生理触发因素