Surface Signatures of Neuronal and Glial Extracellular Vesicles

神经元和胶质细胞外囊泡的表面特征

• 批准号: 10460520
• 负责人: David Aaron Routenberg
• 金额: $ 89.54万
• 依托单位: MESO SCALE DIAGNOSTICS, LLC
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-19 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10460520
• 关键词: Address; Alzheimer' s disease patient; Alzheimer’s disease biomarker; Area; Astrocytes; Automation; Biological Assay; Biological Markers; Blood; Blood Platelets; Body Fluids; Brain; Brain Diseases; Cell Communication; Cell Line; Cell Surface Proteins; Cells; Communication; Consumption; Diagnosis; Endothelium; Epithelial; Goals; Human; Immunoassay; Measurement; Measures; Methods; Microglia; Molecular; National Institute on Aging; Neural Cell Adhesion Molecule L1; Neuraxis; Neurodegenerative Disorders; Neurons; Oligodendroglia; Patients; Performance; Peripheral; Phase; Plasma; Population; Preparation; Procedures; Process; Proteins; Recovery; Role; Saliva; Sampling; Source; Specificity; Standardization; Surface; Techniques; Technology; Time; Urine; base; brain cell; cell type; cost effective; extracellular vesicles; improved; induced pluripotent stem cell; instrument; interest; multiplex assay; nervous system disorder; novel; novel strategies; particle; screening; tau-1

Abstract Extracellular vesicles (EVs) are believed to be an important means of cell-to-cell communication in the central nervous system. Various types of cells in the human brain secrete EVs, which are each likely to have distinct functions. Studying different types of EVs and their roles in the healthy brain and in neurological disease requires a reliable means of capturing these particles from readily accessible body fluids, like blood. Our goal is to develop a new way to capture specific types of EVs based on the molecules present on their surfaces. Our hypothesis is that we can use the presence of two or more specific molecules on the surface of an EV to select EVs secreted by a particular cell type. MSD has previously developed efficient and highly sensitive methods for screening EVs to determine combinations of proteins present on their surfaces. We will use these methods along with samples provided by our collaborators at the National Institute on Aging to identify specific surface-marker signatures for each of the four most common types of cells in the central nervous system. We will develop a new approach to capturing EVs with each of these surface marker signatures. This approach, which will only capture the EVs having all of the targeted surface markers, should greatly improve the capture specificity, thus addressing one of the main shortcomings of the existing CNS EV isolation methods. Greater specificity will enable more targeted studies of the EVs from each CNS cell type than those that are presently performed. We will use this approach to determine which human biofluids can be a reliable source of EVs from the central nervous system and to estimate the level of each type of EV in peripheral body fluids. We will also measure the proteins contained within the EVs, something that requires exceptionally sensitive assays. We recently combined ultrasensitive immunoassays with EV- specific capture methods to enable accurate measurement of specific EV cargo proteins. We will assay the EV cargo for proteins specific to each of the four CNS cell types to help confirm the cellular origin of the isolated EVs. We will also measure known EV-associated biomarkers of Alzheimer’s disease in patient samples to determine in which EV types they are enriched and to see whether measuring the biomarker within a particular type of EV might increase its predictive power. Lastly we plan to develop a fully automated version of the EV isolation and cargo protein assays. This will enable rapid and reliable preparation of EV samples from hundreds of samples at a time, allowing studies that presently would be either impossible or too time- consuming to be cost-effective.

摘要 细胞外囊泡（Extracellular vesicles，EV）被认为是细胞间相互作用的重要手段， 在中枢神经系统的沟通。人类大脑中的各种类型的细胞分泌 电动汽车，每一个都可能有不同的功能。研究不同类型的电动汽车及其 在健康大脑和神经系统疾病中的作用需要一种可靠的手段来捕获 这些颗粒来自于容易接触到的体液，比如血液。我们的目标是开发一种新的方式 基于存在于其表面上的分子来捕获特定类型的EV。我们 假设是，我们可以使用两个或更多个特定分子的表面上的存在， EV以选择由特定细胞类型分泌的EV。 默沙东此前已开发出高效、高灵敏度的EV筛查方法 以确定其表面上存在的蛋白质的组合。我们将使用这些方法 沿着我们的合作者在国家老龄化研究所提供的样本， 中央淋巴结中四种最常见细胞类型的特异性表面标记物特征 神经系统 我们将开发一种新的方法来捕捉电动汽车与每一个这些表面标记 签名.这种方法，这将只捕捉电动汽车具有所有的目标表面， 标记物，应该大大提高捕获特异性，从而解决了一个主要的 现有CNS EV分离方法的缺点。更高的特异性将使更多 与目前进行的那些相比，来自每种CNS细胞类型的EV的靶向研究。 我们将使用这种方法来确定哪些人类生物流体可以成为EV的可靠来源 从中枢神经系统的EV水平，并估计各类型EV在外周体中的水平 流体. 我们还将测量EV中包含的蛋白质，这需要 异常敏感的检测。我们最近将超灵敏免疫测定与EV- 特异性捕获方法使得能够精确测量特异性EV货物蛋白。我们 将分析EV货物中四种CNS细胞类型的特异性蛋白质，以帮助确认 分离的EV的细胞来源。我们还将测量已知的EV相关生物标志物 阿尔茨海默病的患者样本，以确定其中EV类型，他们富集， 为了了解测量特定类型EV中的生物标志物是否可能增加其 预测能力。 最后，我们计划开发一种完全自动化的EV分离和货物蛋白 测定。这将使快速和可靠的准备EV样品从数以百计的 一次采样，允许研究，目前将是不可能的或太长的时间- 消费，才是有成本效益的。