JPND - Intraneuronal transport-related pathways across neurodegenerative diseases

JPND - 跨神经退行性疾病的神经元内转运相关途径

• 批准号: MR/R024782/1
• 负责人: Simon Andrews
• 金额: $ 32.74万
• 依托单位: Babraham Institute
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FR024782%2F1
• 关键词: JPND; Intraneuronal; transport; related; pathways

The overall goal of this proposal is to identify pathways/protein networks underlying the spreading of distinct aggregates of Abeta1-42, Tau, alpha-synuclein or HTTExon 1 involved in Alzheimer disease, Parkinson's disease, and Huntington disease, respectively. The spread of the lesions is a common feature of all these diseases, but strikingly starts and progresses in different brain sub-regions for each pathology. According to recent observations distinct aggregate conformers could be responsible for different clinical subtypes. Here, we aim at revealing the specific mechanisms by which distinct fibrillary protein polymorphs of Abeta1-42, Tau, alpha-synuclein and HTTExon 1, are taken up and transported by neurons. We will use different types of mouse neurons maintained in primary culture to establish a structure-function relationship for distinct pathogenic protein assemblies and their conformers by analyzing their binding, uptake and cellular trafficking after extracellular application. By doing so we will address:By doing so we will address:1. How distinct pathogenic fibrillary protein polymorphs bind to neuronal cells and to which membrane components;2. How exogenous pathogenic fibrillary proteins are taken up and transported;3. How pathogenic fibrillary proteins traffic within cells by identifying;4. The relationship between the traffic of pathogenic fibrillary proteins and neuronal activity;5. The protein networks that account for distinct diseases;6. How genetic risk factors affect the interactomes and transport mechanisms of distinct pathogenic fibrillary proteins polymorphs;7. The identification of common pathways involved as novel therapeutic targets.More specifically, we will* quantify the binding of these distinct pathogenic fibrillary proteins polymorphs to the neuronal plasma membrane;* identify their "receptors" or membrane protein partners;* assess endocytosis;* identify their cargoes within the cells;* quantify intracellular transport and their export/release in the extracellular medium after anterograde or retrograde transport to identify common and divergent protein networks and characteristics that account for distinct diseases.The different partners of this project have been selected for their expertise to cover these specific tasks.* The LAC/ENS Paris-Saclay team will apply its recent intraneuronal transport assay, relying on fluorescent nanodiamond tracking.* Neuro-PSI and CNCR teams have expertise in interaction proteomics to reveal proteins with functional similarities and specificities in the different trafficking steps.* The consortium will also take advantage of novel mouse transgenic LOAD models, PD models and Halo-tagged-knockin models recently obtained and validated by the FP7-HEALTH AgedBrainSYSBIO consortium (coordinated by LAC/ENS Paris-Saclay) to identify common underlying mechanisms linked to intraneuronal transport.* These readouts will be used to build quantitative models of aggregates trafficking and identify common pathways, using the expertise of the BI partner and of the MSSM as external partner.

这项提案的总体目标是确定Abeta1-42、Tau、α-突触核蛋白或HTTExon 1分别与阿尔茨海默病、帕金森病和亨廷顿病相关的不同聚集体扩散的途径/蛋白质网络。病变的扩散是所有这些疾病的共同特征，但在每种病理情况下，病变都在不同的脑区开始和发展。根据最近的观察，不同的聚集体构象可能与不同的临床亚型有关。在这里，我们旨在揭示Abeta1-42、Tau、α-突触核蛋白和HTTExon 1的不同纤维蛋白多态被神经元摄取和运输的具体机制。我们将使用原代培养的不同类型的小鼠神经元，通过分析它们在细胞外应用后的结合、摄取和细胞运输，建立不同致病蛋白组件及其构象的结构-功能关系。通过这样做，我们将解决：1.不同的致病纤维蛋白多态如何与神经细胞结合以及与哪些膜成分结合；2.外源致病纤维蛋白如何被吸收和运输；3.致病纤维蛋白如何通过鉴定在细胞内运输；4.致病纤维蛋白的运输与神经元活动之间的关系；5.不同疾病的蛋白质网络；6.遗传风险因素如何影响不同致病纤维蛋白多态的相互作用和运输机制；7.确定作为新治疗靶点的共同途径。更具体地说，我们将*量化这些不同的致病原纤维蛋白多态与神经细胞膜的结合；*确定它们的“受体”或膜蛋白伙伴；*评估内吞作用；*确定它们在细胞内的运输；*量化顺行或逆行运输后细胞内运输及其在细胞外介质中的输出/释放，以确定导致不同疾病的共同和不同的蛋白质网络和特征。该项目的不同合作伙伴因其专业知识而被选中。*LAC/ENS巴黎-Saclay团队将应用其最新的神经元内运输分析，依赖于荧光纳米钻石追踪。*Neuro-PSI和CNCR团队在相互作用蛋白质组学方面拥有专业知识，以揭示不同运输步骤中具有功能相似性和特异性的蛋白质。*该联盟还将利用新的小鼠转基因负荷模型，PD模型和光环标记敲门模型最近由FP7-Health AgedBrainSYSBIO联盟(由LAC/ENS巴黎-萨克雷协调)获得并验证，以确定与神经元内运输有关的共同潜在机制。*这些读数将用于建立集合体贩运的量化模型，并确定共同路径，使用BI合作伙伴和MSSM作为外部合作伙伴的专业知识。