Directional proteome analysis of extracellular vesicles in AD models

阿尔茨海默病(AD)是一种与和Tau相关的破坏性神经退行性痴呆 目前在美国有540万人患有脑部疾病。AD的主要假设是 一种/淀粉样蛋白假说，该假说认为A的积累是 促进直体病变、线粒体和突触功能障碍的致病进展，以及 神经炎症，导致突触和神经元丢失。新出现的证据表明，释放和 细胞外小泡(EV)从一个细胞到另一个细胞的摄取是细胞间的一种重要形式 可以在不同细胞间传递有益或致病信号的通讯。我们的初步数据 结果表明，A42寡聚体和突变型Tau显著改变EV(外切体& 微泡)。尽管有这些显著的变化，但EV蛋白质组和相互作用的特定变化 而特定神经元室和小胶质细胞之间的定向效应尚不清楚。此外，在给定 EVS在血浆等生物体液中含量丰富，脑源性EVS的蛋白质组发生变化 为从血浆中开发新的生物标记物提供了独特的机会。我们的工作假设是 AD的病理驱动因素显著改变了EV蛋白质组(外切体和MVS)以及它们的 通过局部和非局部机制致病，以及脑EV蛋白质组的这些变化将 作为血浆中有用的病理和疾病特异性生物标志物。在这份建议书中，我们将介绍 并验证神经元/小胶质细胞培养的EV在AD致病反应中蛋白质组的变化 驱动因素和2)确定分离的42处理衍生的外切体的局部和定向致病性 来自神经元和小胶质细胞的相互作用的环境。因此，这些研究的结果将提供关键的 42、Tau和小胶质细胞在EVS和EV释放中的交互和定向作用 蛋白质及其致病性，同时确定AD中EV蛋白质组的主要变化 致病环境。这些信息将有助于识别基于EV的AD生物标记物 并建立了一个广泛的病理来源的EVS及其功能表征的平台 蛋白质组内容。

Alzheimer’s disease (AD) is a devastating neurodegenerative dementia associated with A and Tau pathologies in brain that currently afflicts 5.4 million individuals in the USA. The major hypothesis of AD is the A/amyloid hypothesis, which states that the accumulation of A is an early and necessary event in the pathogenic progression that promotes tauopathy, mitochondrial & synaptic dysfunction, and neuroinflammation, leading to synaptic and neuronal loss. Emerging evidence indicates that the release and uptake of extracellular vesicles (EVs) from one cell to another represent an important form of intercellular communication that could transmit beneficial or pathogenic signals across different cells. Our preliminary data indicate that A42 oligomers and mutant Tau dramatically alter the release of EVs (exosomes & microvesicles). Despite these significant changes, the specific changes in the EV proteome and the interactive and directional effects between specific neuronal compartments and microglia are unknown. Moreover, given that EVs are abundant in biological fluids such as plasma, alterations in the proteome of brain-derived EVs provides the unique opportunity to develop novel biomarkers from blood plasma. Our working hypothesis is that AD pathological drivers significantly alter the EV proteome (exosomes & MVs) as well as their pathogenicity via local and nonlocal mechanisms and that those changes in the EV proteome of brain will serve as useful pathology and disease-specific biomarkers in blood plasma. In this proposal, we will 1) profile and validate the changes in proteome of EVs from neuron/microglia cultures in response to AD pathogenic drivers and 2) determine the local and directional pathogenicity of A42 treatment-derived exosomes isolated from the interactive milieu of neurons and microglia. Therefore, the results of these studies will provide critical insights to the interactive and directional effects of A42, Tau, and microglia in the release of EVs and EV proteins as well as their pathogenicity, while identifying the major changes in the EV proteome in the AD pathogenic setting. Such information collectively will aid in the identification of EV-based biomarkers for AD and establish a platform for extensive functional characterization of pathologically-derived EVs and their proteome content.