JPND - Intraneuronal transport-related pathways across neurodegenerative diseases

本提案的总体目标是确定分别涉及阿尔茨海默病、帕金森病和亨廷顿病的Abeta 1 -42、Tau、α-突触核蛋白或HTTExon 1的不同聚集体的传播的潜在途径/蛋白质网络。病变的扩散是所有这些疾病的共同特征，但对于每种病理，在不同的脑子区域中开始和发展。根据最近的观察，不同的聚集体构象可能导致不同的临床亚型。在这里，我们的目的是揭示特定的机制，其中不同的神经元蛋白多态性的Abeta 1 -42，Tau，α-突触核蛋白和HTTExon 1，采取和运输的神经元。我们将使用不同类型的小鼠神经元保持在原代培养建立一个结构-功能关系，不同的致病性蛋白质组装体和它们的构象，通过分析它们的结合，摄取和细胞外应用后的细胞运输。通过这样做，我们将解决：1。不同的致病性神经元蛋白多态性如何与神经元细胞结合以及与哪些膜成分结合;2.外源性致病蛋白是如何被摄取和转运的;3.如何致病性蛋白质交通细胞内通过识别;4.致病性神经元蛋白的运输与神经元活动之间的关系;5.解释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性糖蛋白多态性的相互作用组和转运机制;7.作为新的治疗靶点的共同途径的鉴定。更具体地说，我们将 * 定量这些不同的致病性神经元蛋白质多态性与神经元质膜的结合;* 鉴定它们的“受体”或膜蛋白伴侣;* 评估内吞作用;* 鉴定它们在细胞内的货物;* 量化细胞内转运及其输出/在顺行或逆行转运后在细胞外介质中释放，以鉴定共同和不同的蛋白质网络和特征，这一项目的不同合作伙伴是根据他们的专门知识来完成这些具体任务的。LAC/ENS Paris-Saclay团队将应用其最新的神经元内转运试验，依赖于荧光纳米金刚石跟踪。Neuro-PSI和CNCR团队在相互作用蛋白质组学方面具有专业知识，可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FP 7-HEALTH脑内SYSBIO联盟（由LAC/ENS Paris-Saclay协调）最近获得并验证的新型小鼠转基因LOAD模型，PD模型和Halo标记的敲入模型，以确定与神经元内运输相关的共同潜在机制。这些读数将用于建立总量贩运的定量模型，并利用BI合作伙伴和MSSM作为外部合作伙伴的专门知识确定共同途径。

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.