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变性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，LRRK2 激酶活性异常是家族性和特发性迟发性帕金森病 (PD) 的一个显着特征。家族性 LRRK2 突变通常与激酶活性升高相关。识别调节 LRRK2 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。 LRRK2 在溶酶体应激源、氯喹 (CO) 和 LLOMe 诱导的扩大的溶酶体中被招募和激活。我们假设 LRRK2 招募到应激溶酶体是溶酶体应激反应的一部分，其作用是将溶酶体恢复到基础状态。 LRRK2 与应激溶酶体的关联通常是短暂的。值得注意的是，LRRK2 与应激溶酶体的关联在 PD 相关的 Vps35-D620N 突变细胞中显着增强。这一发现表明 LRRK2 因 Vps35-D620N 细胞中的溶酶体应激而过度激活。因此，应激诱导的 LRRK2-溶酶体 关联和激活受到严格调节，而这种调节会被导致家族性帕金森病的突变破坏。对 LRRK2-溶酶体关联的其他调节剂进行的小型集中化学抑制剂筛选确定了 PKD。 PKD 与 LRRK2 类似，由溶酶体应激激活，而其失活导致 LRRK2-溶酶体异常关联和激活。该提案旨在鉴定将 LRRK2 招募到应激溶酶体的分子实体，并表征 PKD 信号通路在 LRRK2 激活和致病毒性中的作用。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 的新型调节剂，并确定溶酶体应激作为迟发性 PD 的病理生理触发因素