LRRK2 Biology in Parkinson's Disease

LRRK2 在帕金森病中的生物学作用

基本信息

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
项目类别：
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

项目摘要

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 生物学在帕金森病中的应用帕金森病 (PD) 是一种复杂的神经退行性疾病，具有散发性和家族性。最近显示，富含亮氨酸重复激酶 2 (LRRK2) 的突变可导致 4% 的常染色体显性家族性病例和全球 1% 的散发性病例的临床和病理表型。 LRRK2 PD 患者与经典的迟发性 PD 相似，LRRK2 敲除动物对 α- 具有抵抗力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。该项目将探讨异常激酶活性如何导致疾病的机制。我们的假设是，引起 LRRK2 GS 的疾病通过致病性引起翻译失调。 s15 磷酸化，导致蛋白质表达改变以及神经元功能障碍和死亡。全面了解 mRNA 翻译、转录组和转录组的分子变化 LRRK2 GS 引发的蛋白质组是了解 DA 漏洞所必需的。还有一个重要的因素。与 α-突触核蛋白的相互作用会影响 DA 脆弱性，但其机制尚不清楚。目标 1 解决关于增加激酶活性的重要性的持续争议 LRRK2 GS 突变对 DA 神经元活力的影响，制备了 LRRK2 GS 激酶死亡（LRRK2 GS/DA）小鼠。这些小鼠还提供了一个模型来探索 LRRK2 的非激酶作用，而这是随着时间的推移，将监测行为、神经解剖学和神经化学变化。 LRRK2 和 α-突触核蛋白毒性和传播之间的相互作用将在 LRRK2 KO 和确定 LRRK2 功能丧失的转基因品系是保护性的，而 LRRK2 功能获得的则是毒性的。目标 2 将解决磷酸化 s15 是 LRRK2 GS 致病靶点的新观察结果，将在转基因小鼠和人类死后组织中监测 s15 和磷酸化 s15 的表达。将确定磷酸模拟物 s15 是否足以引起神经变性以及磷酸化缺陷 s15 是否足以引起神经变性这些研究可以在全局和空间限制模型中提供针对 LRRK2 GS 神经毒性的保护。将在体内确认s15是否是LRRK2 GS的致病底物。目标 3 将定义 mRNA 翻译、转录组和蛋白质组的具体变化，使用基因工程小鼠和人类多巴胺能培养物中的先进技术的使用。小鼠和人类模型可以在完整的大脑中进行评估，同时研究导致的急性变化相关人类系统中的慢性神经退行性疾病的无偏见综合数据集将是一个对所有 PD 调查人员来说都是宝贵的资源。该项目的目标是识别神经退行性变信号级联中的节点，这些节点可以为PD治疗策略的未来发展提供新的靶点。