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: 10019714
• 负责人: David Aaron Routenberg
• 金额: $ 89.38万
• 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 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10019714
• 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; Epithelium; 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; nervous system disorder; novel; novel strategies; particle; screening; tau-1

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.

细胞外小泡(EVS)被认为是细胞间的重要途径 中枢神经系统中的交流。人脑中的各种类型的细胞分泌 电动汽车，每一种都可能具有不同的功能。研究不同类型的电动汽车及其 在健康的大脑和神经疾病中的作用需要一种可靠的手段来捕捉 这些微粒来自容易接触到的体液，比如血液。我们的目标是开发一种新的方式 根据电动汽车表面存在的分子捕捉特定类型的电动汽车。我们的 假设我们可以利用两个或更多特定分子在表面上的存在 EV用于选择由特定细胞类型分泌的EV。 MSD此前已开发出高效且高度敏感的电动汽车筛查方法 以确定存在于其表面的蛋白质的组合。我们将使用这些方法 以及我们在国家老龄化研究所的合作者提供的样本，以确定 中央四种最常见的细胞类型中每一种的特定表面标记特征 神经系统。 我们将开发一种新的方法来捕捉电动汽车的每一个表面标记 签名。此方法将仅捕获具有所有目标表面的电动汽车 标记物，应大大提高捕获的特异性，从而解决主要问题之一 现有中枢神经系统EV隔离方法的不足之处。更大的专用性将带来更多 对每种中枢神经系统细胞类型的EVS进行有针对性的研究，而不是目前进行的那些研究。 我们将使用这种方法来确定哪些人体体液可以作为肠道病毒的可靠来源。 并估计每种EV在外周体内的水平 体液。 我们还将测量电动汽车中包含的蛋白质，这需要 异常敏感的化验结果。我们最近将超灵敏的免疫分析与EV- 特定的捕获方法，能够准确测量特定的EV货物蛋白。我们 将分析EV货物中四种CNS细胞类型中每一种的特定蛋白质，以帮助确认 孤立电动汽车的细胞起源。我们还将测量已知的EV相关生物标记物 以确定哪些EV类型富含阿尔茨海默病 来看看在特定类型的电动汽车中测量生物标记物是否会增加其 预测能力。 最后，我们计划开发一种全自动版本的EV分离和货物蛋白 化验。这将使快速和可靠地从数百个EV样本中制备 一次采样，允许进行目前不可能或时间太长的研究- 消费以达到成本效益。