Dissecting signaling pathways and seeking EV phosphoproteins as novel biomarkers for Alzheimer's Disease

项目摘要 阿尔茨海默病（AD）是一种进行性和致命的神经退行性疾病，表现为 认知和记忆衰退。AD的特征性病理变化是纤维蛋白 在大脑皮层中沉积，可能通过在细胞间隙中沉积β-淀粉样蛋白（AE）， 在细胞中过度磷酸化Tau蛋白。然而，确切的分子机制和致病性 AD的信号传导尚不清楚，研究人员一直在寻找新的线索和可靠的 诊断AD。在这项专注于创新和转化AD研究的R 01研究中，我们将 介绍基本疾病机制和临床观点前沿的新策略。 本计画的长期目标是：1）发展系统的策略来剖析激酶-底物 与AD发病相关的信号网络，重点是识别和验证 AD关键致病途径中的直接激酶-底物关系; 2）发展 血浆细胞外囊泡（EV）中的磷酸化蛋白用于潜在的临床诊断。作为 磷酸化在疾病如AD、EV、糖尿病和糖尿病的早期发病和进展中起主要作用， 磷蛋白有望成为积极追求的目标，作为细胞状态的指标 以及用于体外疾病诊断。我们将整合新的蛋白质组学方法来识别AE， 与AD发病机制相关的Tau上游激酶，并剖析AE和Tau- 相关的信令网络。策略，荧光互补质谱 （FCMS），将利用蛋白质互补和定量蛋白质组学建立一个高水平的 通过筛选方法来鉴定AE和Tau的直接上游激酶， AD进展。因此，我们将实现以下具体目标：1）：了解 通过构建高分辨率激酶-底物网络驱动AD进展 在Aβ和Tau相关信号通路中的作用; 2）：建立用于 血浆EV中已知AD生物标志物的靶向检测;和3）：磷蛋白的发现 作为AD检测和监测的新型生物标志物。

PROJECT SUMMARY Alzheimer's disease (AD) is a progressive and fatal neurodegenerative disorder manifested by cognitive and memory deterioration. The characteristic pathology changes in AD are fibrin deposition in cerebral cortex, likely through the deposition of beta-amyloid (AE) in cell space and hyperphosphorylated Tau protein in cell. However, the exact molecular mechanism and pathogenic signaling of AD is not clear and researchers have been searching for new leads and reliable diagnosis for AD. In this R01 study that focuses on innovative and translational AD research, we will introduce novel strategies at the forefront of basic disease mechanism and clinical perspectives. The long term goal of this project is 1) to develop systematic strategies to dissect kinase-substrate signaling network related to onset of AD, with an emphasis on the identification and validation of direct kinase-substrate relationship in AD's critical pathogenic pathways; and 2) to develop phosphorylated proteins in plasma extracellular vesicles (EVs) for potential clinical diagnosis. As phosphorylation is a major player in early onset and progression of diseases such as AD, EV phosphoproteins are expected to become actively pursued targets as indicators of cellular states and for in vitro disease diagnosis. We will integrate novel proteomic approaches to identify AE and Tau upstream kinases associated with the pathogenesis of AD and to dissect AE and Tau- associated signaling networks. The strategy, fluorescence complementation mass spectrometry (FCMS), will utilize protein complementation and quantitative proteomics to establish a high throughput screening method to identify direct upstream kinases of AE and Tau associated with the AD progression. Accordingly, we will achieve the following specific aims: 1): Understanding the driving force of AD progression through the construction of high resolution kinase-substrate network in Aβ and Tau associated signaling pathways; 2): Establishment of an analytical platform for targeted detection of known AD biomarkers in plasma EVs; and 3): Discovery of phosphoproteins from plasma EVs as novel biomarkers for AD detection and monitoring.