Effect of agent and host factors on alpha-synuclein strain pathogenesis

病原体和宿主因素对α-突触核蛋白菌株发病机制的影响

• 批准号: 10754428
• 负责人: Amanda L. Woerman
• 金额: $ 11.5万
• 依托单位: UNIVERSITY OF MASSACHUSETTS AMHERST
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-15 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10754428
• 关键词: Affect; American; Anatomy; Area; Axonal Transport; Binding; Biochemical; Biological; Biological Assay; Biology; Brain; Cell Line; Cell model; Central Nervous System; Chemicals; Chimeric Proteins; Clinical; Disease; Disease Progression; Dynein ATPase; Exhibits; Genetic; Goals; Heterogeneity; Human; In Vitro; Injections; Integration Host Factors; Investigation; Knowledge; Label; Mammalian Cell; Maps; Microtubule Polymerization; Modeling; Molecular; Molecular Conformation; Motor Activity; Movement Disorders; Multiple System Atrophy; Mus; Neural Pathways; Neuroanatomy; Neurodegenerative Disorders; Neurons; Parkinson Disease; Pathogenesis; Pathway interactions; Patients; Pattern; Peripheral; Prions; Property; Proteins; Recombinants; Research; Role; Route; Sampling; Shapes; Source; Structure; Sucrose; Symptoms; Synapses; Testing; Therapeutic; Therapeutic Intervention; Tissues; Transgenic Mice; Work; alpha synuclein; biophysical properties; clinical heterogeneity; conformer; disease heterogeneity; disease phenotype; experimental study; in vivo; in vivo imaging; innovation; insight; mouse model; nerve transection; nervous system disorder; neuropathology; pre-formed fibril; prion-like; protein misfolding; sciatic nerve; skills; synucleinopathy; tool; transmission process; transport inhibitor

Protein misfolding diseases, or proteinopathies, are a group of invariably fatal neurodegenerative disorders affecting more than 6.8 million Americans. In multiple system atrophy (MSA) and other synucleinopathy patients, the protein α-synuclein (α-syn) misfolds into a self-templating conformation that spreads via a prion- like manner throughout the body, including the central nervous system (CNS). It is hypothesized that the conformation, or strain, that α-syn misfolds into encodes information about the clinical symptoms and neuropathologies a patient will develop. While previous studies focused on the biochemical differences between α-syn strains, the mechanism of how those differences encode distinct biological phenotypes of disease is poorly understood. The long-term goal of our research is to identify the agent and host factors that contribute to the varied clinical presentations observed across synucleinopathies. In this proposal, we will test the hypothesis that strain-specific differences in aggregate transport and neuroanatomical spread contribute to disease pathogenesis. In Aim 1, we will use alexa fluor-labeled α-syn aggregates to investigate the rate and direction of axonal transport in vitro and in vivo. To determine the molecular mechanisms responsible for α-syn transport, we will use chemical and genetic tools to disrupt microtubule polymerization, dynein motor activity, and dynein cargo adaptor binding, and quantify the strain-specific effects on α-syn axonal transport. In Aim 2, we will determine the role of trans-synaptic spread on α-syn strain pathogenesis. To rigorously perform these studies, we will first determine the titer of three different α-syn strains both in vitro and in vivo. We will then use the sciatic nerve injection model, with and without nerve transection, to determine if α-syn neuroinvasion relies exclusively on trans-synaptic spread, of if extraneural pathways contribute to disease pathogenesis when the same titer of each strain is injected. Finally, we will perform a thorough disease pathogenesis study to establish a temporal-spatial map of strain-specific α-syn spread. This work is innovative because it is the first study to investigate how interactions between the host and strain impact disease progression, and to establish between in vitro and in vivo α-syn titers. This work is significant because it is the first to investigate how interactions between host and strain contribute to the mechanisms underlying axonal transport and trans-synaptic spread of disease. Critically, by identifying the cellular and molecular machinery responsible for α-syn propagation, the results of these experiments will lead to new areas of promising investigation.

蛋白质错误折叠疾病或蛋白质病是一组不变的致命神经退行性疾病 影响超过680万美国人。在多系统萎缩（MSA）和其他突触核酸 患者，蛋白质α-突触核蛋白（α-syn）不折叠成一个自我鉴定构象，该构象通过prion- 整个身体的方式，包括中枢神经系统（CNS）。假设 α-syn错误折叠中的构象或应变编码有关临床符号和的信息 神经病理学患者会发育。而先前的研究集中于生化差异 在α-syn菌株之间，这些差异如何编码不同的生物学表型的机制 疾病知之甚少。我们研究的长期目标是确定代理和寄宿因素 有助于跨突触核心病变观察到的各种临床表现。在此建议中，我们将测试 骨料转运和神经解剖学扩散的应变特异性差异的假设有助于 疾病发病机理。在AIM 1中，我们将使用Alexa荧光标记的Alpha-Syn聚集体来研究速率和 轴突运输的方向在体外和体内。确定负责α-syn的分子机制 运输，我们将使用化学和遗传工具破坏微管聚合，动力蛋白运动活动， 和Dynein货物适配器结合，并量化应变特异性对α-Syn轴突运输的影响。在AIM 2中， 我们将确定跨突触扩散在α-Syn菌株发病机理上的作用。严格执行这些 研究，我们将首先确定体外和体内三种不同α-Syn菌株的滴度。然后我们将使用 坐骨神经注射模型，有或没有神经传播，以确定α-syn神经浸觉是否依赖 if外部途径仅在跨突触传播上，当 注入每个应变的相同滴度。最后，我们将进行彻底的疾病发病机理研究，以建立 应变特异性α-SYN扩散的临时空间图。这项工作具有创新性，因为它是首次研究 研究宿主与菌株之间的相互作用如何影响疾病的进展，并在 体外和体内α-syn滴度。这项工作很重要，因为它是第一个研究互动方式 宿主和应变之间有助于轴突运输和反式突触的基础机制 疾病。至关重要的是，通过识别负责α-syn繁殖的细胞和分子机械， 这些实验的结果将导致有望调查的新领域。