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.

项目概要-项目4：帕金森病中的LRRK 2生物学 帕金森病（PD）是一种复杂的神经退行性疾病，既有散发性，又有家族性。 富含亮氨酸重复序列激酶2（LRRK 2）的突变最近已被证明可导致4%的常染色体 占全球主要的家族性病例和1%的散发病例。的临床和病理表型 LRRK 2 PD患者与经典的晚发型PD相似，LRRK 2敲除动物对α- 突触核蛋白多巴胺能（DA）神经变性，进一步强调了该基因的潜在重要性。 这个项目将解决激酶活性异常如何导致疾病的机制。 我们的假设是，引起疾病的LRRK 2 GS通过致病性引起翻译失调 S15的磷酸化，导致改变的蛋白质表达和神经元功能障碍和死亡。 全面了解mRNA翻译、转录组和基因组中的分子变化， LRRK 2 GS引起的蛋白质组是理解DA脆弱性所必需的。还有一个重要 与α-突触核蛋白的相互作用影响DA的脆弱性，但机制尚不清楚。 目的1：解决目前关于增加激酶活性的重要性的争论， 制备LRRK 2 GS突变对DA神经元活力的影响、LRRK 2 GS激酶死亡（LRRK 2 GS/DA）小鼠。 这些小鼠还提供了一个模型，以探索LRRK 2的非激酶作用，这是缺乏的。 领域随着时间的推移，将监测行为、神经解剖学和神经化学变化。功能 将在LRRK 2 KO中监测LRRK 2和α-突触核蛋白毒性和传播之间的相互作用， 转基因系的LRRK 2功能丧失是保护性的，而LRRK 2功能获得是毒性的。 目的2将阐述磷酸化s15是LRRK 2 GS的致病靶点的新观察结果， 将在转基因小鼠和人死后组织中监测S15和磷酸-S15的表达。它 如果磷酸化模拟S15足以引起神经变性和如果磷酸化缺陷S15 可以在全局和空间限制模型中提供免受LRRK 2 GS神经毒性的保护。这些研究 将在体内证实s15是否是LRRK 2 GS的致病底物。 目的3将明确mRNA翻译，转录组和蛋白质组的具体变化， 基因工程小鼠和人类多巴胺能培养物的先进技术。两者的使用 小鼠和人类模型允许在完整的大脑中进行评估，同时研究导致 与慢性神经退化的关系无偏见的综合数据集将是一个 所有警局调查员的宝贵资源 该项目的目标是确定神经退行性疾病信号级联中的节点， 为未来开发治疗PD的治疗策略提供了新的靶点。