Development of A Novel Magnetic Exosomal Assay for Diagnosis of Diseases with Impaired Vascular Integrity

 描述（由申请人提供）：该小型企业创新研究第1阶段项目旨在开发和表征一种新型磁性外泌体测定法，以分离和检测人血清中单个完整外泌体浓度。这项拟议的研究还将评估磁性外泌体测定对血管完整性受损疾病的诊断潜力。血管完整性或血管连续性的维持是通过内皮细胞-细胞连接维持的基本过程。内皮屏障功能的缺陷是几种疾病过程的起始因素，包括动脉粥样硬化、缺血/再灌注、肿瘤血管生成、癌症转移、糖尿病、败血症和急性肺损伤。血管完整性受损的初始事件是来自内皮细胞（EC）的称为外泌体的新型细胞外囊泡（EV）的急剧增加。因此，开发高度灵敏、可重复且易于实施的检测复杂临床样品中外来体浓度的测定法可具有潜在的诊断和治疗益处。在本研究提案中，我们将开发和表征一种新型磁性外泌体检测试剂盒和读数器，用于快速分离和检测人血清中单个完整外泌体浓度。此外，我们将评估社区获得性肺炎患者血清的优化磁性外泌体检测。本研究的最终目标是开发一种高度敏感、可重复和易于实施的外泌体检测方法，作为诊断各种类型血管完整性受损相关疾病患者的工具。

 DESCRIPTION (provided by applicant): This Small Business Innovation Research Phase 1 project aims to develop and characterize a novel Magnetic Exosomal Assay to isolate and detect single intact exosome concentrations in human serum. This proposed research will also evaluate the diagnostic potential of the Magnetic Exosomal Assay for diseases with impaired vascular integrity. Vascular integrity or the maintenance of blood vessel continuity is a fundamental process maintained through endothelial cell-cell junctions. Defects in endothelial barrier function are an initiating factor in several disease processes including atherosclerosis, ischemia/reperfusion, tumor angiogenesis, cancer metastasis, diabetes, sepsis and acute lung injury. An initial event in impaired vascular integrity is a dramatic increase in a novel type of extracellular vesicle (EV) from endothelial cells (EC) called exosomes. Therefore, the development of a highly sensitive, reproducible and easy-to- implement assay to detect exosome concentrations in complex clinical samples can have potential diagnostic and therapeutic benefit. In this research proposal, we will develop and characterize a novel Magnetic Exosomal Assay kit and reader for fast isolation and detection of single intact exosome concentrations in human serum. In addition, we will assess the optimized Magnetic Exosomal Assay on Community-Acquired Pneumonia patient serum. The ultimate goal of this research is to develop a highly sensitive, reproducible and easy to implement exosomal assay as a diagnostic tool for patients with various types of impaired vascular integrity-related diseases.