LRRK2 Biology in Parkinson's Disease

LRRK2 在帕金森病中的生物学作用

• 批准号: 8882848
• 负责人: VALINA L. 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/8882848
• 关键词: Acute; Address; Affect; Animals; Autopsy; Behavior; Behavioral; Biology; Brain; Cell Death Signaling Process; Cell Survival; Cessation of life; Chronic; Clinical; Complex; Data; Data Set; Development; Disease; Event; Future; Gene Expression Profile; Gene Mutation; Genes; Genetic; Genetic Translation; Genetically Engineered Mouse; Goals; Homeostasis; Human; Investigation; Knock-out; Knowledge; Lead; Link; Longevity; Measures; Mediating; Modeling; Molecular; Monitor; Mus; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neuronal Dysfunction; Neurons; Nodal; Outcome; Parkinson Disease; Pathologic; Pathology; Patients; Phenotype; Phosphorylation; Phosphotransferases; Physiological; Proteins; Proteome; Proteomics; RNA; Research; Research Personnel; Resistance; Resources; Ribosomal Proteins; Role; Signal Transduction; System; TNFRSF5 gene; Technology; Tertiary Protein Structure; Time; Tissues; Toxic effect; Transgenic Mice; Transgenic Organisms; Translations; Viral; age related; alpha synuclein; biological adaptation to stress; dopaminergic neuron; genetic manipulation; in vivo; insight; leucine-rich repeat kinase 2; mimetics; mouse LRRK2 protein; mutant; neurochemistry; neuron loss; neuronal survival; neurotoxicity; novel; protein expression; protein protein interaction; ribosomal protein S15; therapeutic development; transmission process; treatment strategy

PROJECT SUMMARY - PROJECT 4: LRRK2 BIOLOGY IN PARKINSON'S DISEASE Parkinson's disease (PD) is a complex neurodegenerative disorder that is both sporadic and familial. Mutations in the leucine-rich repeat kinase 2 (LRRK2) have recently been shown to result in 4% of autosomal dominant familial cases and 1% of sporadic cases worldwide. The clinical and pathological phenotypes of LRRK2 PD patients are similar to classic late-onset PD and LRRK2 knock out animals are resistant to α- synuclein dopaminergic (DA) neurodegeneration, further emphasizing the potential importance of this gene. This project will address mechanisms of how aberrant kinase activity leads to disease. Our hypothesis is that disease causing LRRK2 GS elicits translational deregulation through pathogenic phosphorylation of s15, leading altered protein expression and neuronal dysfunction and death. Comprehensive understanding of the molecular changes in mRNA translation, the transcriptome and the proteome elicited by LRRK2 GS is required to understand DA vulnerability. There is also an important interaction with α-synuclein that impacts DA vulnerability but the mechanism is not yet known. Aim 1 to address the ongoing controversy regarding the importance of the increased kinase activity of LRRK2 GS mutation on DA neuronal viability, LRRK2 GS kinase-dead (LRRK2 GS/DA) mice were made. These mice also provide a model to explore the non-kinase actions of LRRK2 that has been lacking from the field. Behavioral, neuroanatomical and neurochemical changes will be monitored over time. The functional interaction between LRRK2 and α-synuclein toxicity and transmission will be monitored in LRRK2 KO and transgenic lines to determine loss of LRRK2 function is protective and gain of LRRK2 function is toxic. Aim 2 will address the new observation that phosphorylated s15 is a pathogenic target of LRRK2 GS, expression of s15 and phospho-s15 will be monitored in transgenic mice and human postmortem tissue. It will be determined if phosphomimetic s15 is sufficient to elicit neurodegeneration and if phosphodeficient s15 can provide protection from LRRK2 GS neurotoxicity in global and spatially restricted models. These studies will confirm in vivo whether s15 is a pathogenic substrate of LRRK2 GS. Aim 3 will define the specific changes in mRNA translation, the transcriptome and proteome using advanced technologies in genetically engineered mice and human dopaminergic cultures. The use of both mouse and human models allows assessments in the intact brain while investigating acute changes that lead to chronic neurodegeneration in a relevant human system. The unbiased comprehensive datasets will be a valuable resource to all PD investigators. The goal of this project is to identify nodal points in the signal cascade of neurodegeneration that can provide new targets for future development of therapeutic strategies for the treatment of PD.

项目摘要 - 项目4：LRRK2帕金森氏病生物学 帕金森氏病（PD）是一种复杂的神经退行性疾病，既是零星又是家庭。 最近已显示出富含亮氨酸的重复激酶2（LRRK2）中的突变可导致4％的常染色体 全球占主导地位的家庭病例和1％的零星病例。的临床和病理表型 LRRK2 PD患者类似于经典的后期PD，LRRK2敲除动物对α-具有抗性 综合蛋白多巴胺能（DA）神经变性，进一步强调了该基因的潜在重要性。 该项目将解决异常激酶活性如何导致疾病的机制。 我们的假设是导致LRRK2 GS的疾病通过致病性引起翻译失调 S15的磷酸化，蛋白质表达和神经元功能障碍和死亡的磷酸化。 对mRNA翻译，转录组和转录组的分子变化的全面了解 需要LRRK2 GS引起的蛋白质组才能理解DA脆弱性。还有一个重要的 与影响DA脆弱性的α-核蛋白的相互作用，但尚不清楚该机制。 目的1解决有关增加的激酶活性重要性的争议 在DA神经元活力上，LRRK2 GS激酶（LRRK2 GS/DA）小鼠的LRRK2 GS突变。 这些小鼠还提供了一个模型来探索LRRK2的非激酶动作，而LRRK2缺乏 场地。行为，神经解剖学和神经化学变化将随着时间的流逝而受到监测。功能 LRRK2和α-突触核蛋白毒性和传播之间的相互作用将在LRRK2 KO中进行监测，并且 确定LRRK2功能丧失的转基因线受到保护，LRRK2功能的增益有毒。 AIM 2将解决新的观察结果，即磷酸化的S15是LRRK2 GS的致病靶标， S15和磷酸-S15的表达将在转基因小鼠和人类后组织中进行监测。它 将确定磷酸化S15是否足以引起神经变性和磷缺陷S15 可以在全球和空间限制模型中提供免受LRRK2 GS神经毒性的保护。这些研究 将在体内确认S15是否是LRRK2 GS的致病性底物。 AIM 3使用使用mRNA翻译，转录组和蛋白质组的特定变化 基因工程小鼠和人类多巴胺能培养物中的先进技术。两者的使用 小鼠和人类模型允许评估完整的大脑，同时研究引导的急性变化 在相关的人类系统中进行慢性神经退行性。公正的综合数据集将是 对所有PD调查人员的宝贵资源。 该项目的目的是识别神经变性信号级联中的节点点可以 为未来的理论策略提供了新的目标，以治疗PD。