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

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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.