Developing EV surface proteins as biosignatures for Alzheimer's disease (AD)

项目摘要 越来越多的美国人目前生活与阿尔茨海默氏病（AD）的需求迫切 治疗和诊断解决方案。共识是，早期发现对延迟和 更好地管理疾病。细胞外囊泡（EVs）的最新发现及其潜在的 神经元-神经胶质细胞通讯、突触可塑性或作为 内源性大麻素载体已将其作为神经元疾病诊断的有趣来源 和治疗学。越来越多的功能研究提供了强有力的证据， 基于EV的疾病标志物，如活性miRNA和信号分子，可以很好地识别 在症状发作或疾病的生理检测之前， 早期AD检测的来源。然而，与基于电动汽车的 生物标志物的发现极大地限制了其在临床研究中的应用。在这项STTR研究中， 专注于创新的EV研究和AD检测，我们将应用一种新的EV策略， 分离和分析，严格表征疾病相关EV表面蛋白，然后集中 开发EV表面蛋白作为AD的生物特征。EV表面蛋白，包括 受体、转运蛋白、通道和酶是潜在的诊断生物标志物的来源， 疾病和治疗目标。更重要的是，EV表面蛋白的检测可以是 无需长时间的样品制备程序，如裂解，蛋白质提取， 消化，富集等。所提出的策略引入了一种新的平台技术， EV分离和AD特异性EV表面蛋白的珠上检测。因此， 目标1：实施快速有效的电动汽车综合战略 分离和货物提取;目标#2：与AD连接的EV表面蛋白的表征;目标#3： 开发EV表面蛋白作为AD生物特征的珠上检测。通过完成 本研究的目的是建立一个检测EV表面蛋白的平台技术 作为有前景的AD生物标志物用于进一步验证。

PROJECT SUMMARY The rising number of Americans currently living with Alzheimer’s disease (AD) demands pressing therapeutic and diagnostic solutions. The consensus is that early detection is critical to delay and better manage the disease. The recent discovery of extracellular vesicles (EVs) and their potentially important cellular functions in neuronal-glial communication, synaptic plasticity, or as endocannabinoid carriers has presented them as intriguing sources for neuronal disease diagnosis and therapeutics. The growing body of functional studies have provided strong evidence that EV-based disease markers, such as active miRNAs and signaling molecules, can be identified well before the onset of symptoms or physiological detection of diseases, making them a promising source for early-stage AD detection. However, multiple technical challenges related to EV-based biomarker discovery have greatly limited its applications in clinical studies. In this STTR study that focuses on innovative EV research and detection for AD, we will apply a novel strategy for EV isolation and analysis, rigorously characterize disease-relevant EV surface proteins, and then focus on developing EV surface proteins as biosignatures for AD. EV surface proteins, consisting of receptors, transporters, channels, and enzymes, are a source of potential diagnostic biomarkers of disease and therapeutic targets. More importantly, the detection of EV surface proteins can be achieved in situ without long sample preparation procedure such as lysis, protein extraction, digestion, enrichment and so on. The proposed strategy introduces a novel platform technology for EV isolation and on-bead detection of EV surface proteins specific to AD. Accordingly, the following aims will be completed: Aim #1: Implementation of an integrated strategy for fast and effective EV isolation and cargo extraction; Aim #2: Characterization of EV surface proteins linked to AD; Aim #3: Development of on-bead detection of EV surface proteins as biosignatures for AD. By the completion of this exploratory project, we expect to establish a platform technology to detect EV surface proteins as promising AD biomarkers for further validation.