The impact of synaptic vesicle-binding of alpha-synuclein on neuron function and neuropathology

α-突触核蛋白突触小泡结合对神经元功能和神经病理学的影响

基本信息

批准号：
10406166
负责人：
Jacqueline Burre
金额：
$ 51.26万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-06-01 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10406166
关键词：
Area Basic Science Behavior Behavioral Assay Binding Biochemical Biochemistry Biological Brain Cells Cellular biology Cessation of life Cognitive Complex Data Dependence Development Disease Disease Progression Dose Electrostatics Exhibits Fostering Functional disorder Genes Genetic Polymorphism Histologic Hydrophobic Interactions Impairment In Vitro Injections Knockout Mice Lentivirus Vector Lewy Bodies Lewy Body Dementia Link Liposomes Location Mediating Medical Membrane Mission Molecular Molecular Chaperones Motor Mus Mutation Nerve Neurologic Neurons Onset of illness Parkinson Disease Pathogenesis Pathogenicity Pathologic Pathology Patients Physiological Process Proteins Public Health Published Comment Publishing Rattus Recombinants Research SNAP receptor Substantia nigra structure Synapses Synaptic Vesicles System Techniques Testing Therapeutic Intervention Toxic effect Translational Research United States National Institutes of Health Variant Vesicle Work alpha synuclein alpha synuclein gene base biophysical techniques brain tissue dopaminergic neuron improved in vivo innovation insight interdisciplinary approach loss of function mouse model mutant nervous system disorder neuropathology neurotoxicity neurotransmitter release nigrostriatal degeneration nonhuman primate novel premature rational design single molecule synuclein synucleinopathy tool trafficking treatment strategy vesicular release

项目摘要

Alpha-synuclein (aSyn) pathology is linked to synucleinopathies including Parkinson's disease and Lewy body dementia, but the underlying disease mechanisms remain poorly understood. The prevalent viewpoint has emerged that aggregation of aSyn triggers neuropathology through a gain-of-toxic-function mechanism, and approaches to eliminate aSyn represent an active area of research for treatment. Yet, aSyn aggregation may also endanger neurons by removing aSyn from synaptic vesicles (its physiologically relevant intracellular location) and thereby causing loss-of-function. Through its synaptic vesicle-bound state, aSyn regulates synaptic vesicle trafficking, and chaperones SNARE-complex assembly to maintain neurotransmitter release. Thus, removing aSyn from neurons may not be protective, but detrimental. The objective in this application is to determine the impact of synaptic vesicle-binding of aSyn on aSyn function and neuron survival, using rationally designed variants of aSyn that stabilize synaptic vesicle-binding. The central hypothesis is that stabilizing binding of aSyn on synaptic vesicles reduces aSyn toxicity and pathology. Guided by strong preliminary data, this hypothesis will be tested in three specific aims: 1) Determine the effect of increased synaptic vesicle-binding of aSyn on SNARE-complex assembly; 2) Assess the effect of increased synaptic vesicle-binding of aSyn on synaptic vesicle cycling; and 3) Test if increased synaptic vesicle-binding of αSyn rescues neurotoxicity and pathology in vivo. Under the first aim, SNARE-complex assembly will be quantified in vivo and in vitro, using cell biological and biochemical techniques. Under the second aim, αSyn multimerization, synaptic vesicle pools and clustering, and synaptic vesicle cycling will be quantified, using cell biological, biochemical and biophysical techniques. Under the third aim, mouse models will be generated by stereotactic injections of lentiviral vectors into the substantia nigra of aSyn knockout mice to assess effects of mutant aSyn variants on αSyn-induced toxicity and pathology, using behavioral assays on mice and biochemical, histological and ultrastructural analyses on injected brains. The study is expected to show improved aSyn function and delayed pathology upon stabilization of synaptic vesicle-binding of αSyn. This research is innovative because it 1) tests the novel hypothesis that stabilizing synaptic vesicle-bound αSyn reduces aSyn pathology, 2) creates new tools to study function and dysfunction of αSyn, and 3) uses a multidisciplinary approach to test our hypothesis from single molecules and cellular systems to live mice. This work is significant, because it will 1) clarify the importance of synaptic vesicle-binding of aSyn for neuron function, 2) provide new insights into the molecular mechanism of synaptic vesicle-binding of αSyn, 3) uncover the contribution of loss-of-function of aSyn to disease pathogenesis, and 4) have translational importance for the development of new treatment strategies aimed at stabilizing synaptic vesicle-bound αSyn.

α-突触核蛋白（ASYN）病理学是包括帕金森氏病和路易身体的联系痴呆症，但潜在的疾病机制仍然很差。出现了ASYN的聚集，通过激素功能机制触发神经病理学，并消除Asyn的方法代表了一个积极的治疗领域。还通过从突触速率中取出ASYN来危害神经元位置），从而导致其突触囊泡结合状态。突触囊泡运输和伴侣snare复合物组件以维持神经递质的释放。因此，从神经元中删除ASYN可能不是保护性的，但该应用程序中的目标是有害的。确定ASYN的突触囊泡结合对ASYN功能和神经元存活的影响理性设计的Asyn tutabilize突触囊泡结合的变体。在强者的指导下。预启示数据，该假设将以三个特定目的进行检验：1）确定增加的影响 ASYN在圈圈组件上的突触囊泡结合； 2）评估突触增加的效果 Asyn在突触囊泡循环上的囊泡结合；在第一个目标下，在体内营救神经毒性和病理使用细胞生物学和生化技术，体内和体外。将使用细胞来量化多聚合，突触囊泡池和聚类和突触囊泡循环生物学，生化和生物物理技术。立体定义构成现象的病毒向量进入Asyn淘汰小鼠的底座，以评估影响的影响使用小鼠的行为测定法和αSyn诱导的二氧毒性和病理的突变体变体对注射大脑的生化，组织学和超时期分析。在αSyn的突触囊泡结合稳定后，改善了Asyn功能和延迟病理。研究是创新的，因为它1）检验了稳定突触囊泡结合αSyn的新假设减少ASYN病理学，2）创建新的工具来研究αSyn的功能和功能障碍，3）使用多学科的方法来检验我们从单分子和细胞系统到活小鼠的假设。工作意义重大，因为它会1）崎功能，2）提供有关αSyn突触囊泡结合的分子机制的新见解，3） ASYN对疾病发病机理的功能丧失的贡献和4）对旨在稳定突触囊泡结合的αSyn的新治疗策略的发展。