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

 描述(由申请人提供)：这个小型企业创新研究第一阶段项目旨在开发和表征一种新的磁性外切体分析方法，以分离和检测人血清中单个完整的外切体浓度。这项拟议的研究还将评估磁性胞外体分析对血管完整性受损疾病的诊断潜力。血管完整性或维持血管连续性是通过内皮细胞-细胞连接维持的一个基本过程。内皮屏障功能缺陷是动脉粥样硬化、缺血/再灌注、肿瘤血管生成、肿瘤转移、糖尿病、败血症和急性肺损伤等多种疾病过程的始动因素。血管完整性受损的最初事件是来自内皮细胞(EC)的一种新型细胞外小泡(EV)急剧增加，称为外切体。因此，发展一种灵敏度高、重复性好、易于实现的检测方法来检测复杂临床样本中的外切体浓度，可能具有潜在的诊断和治疗益处。在这项研究计划中，我们将开发和表征一种新型的磁性外切体检测试剂盒和读取器，用于快速分离和检测人血清中单个完整的外切体浓度。此外，我们还将评估优化后的社区获得性肺炎患者血清磁性胞外体检测方法。这项研究的最终目标是开发一种高度敏感、可重复性和易于实施的外体检测方法，作为各种类型血管完整性受损相关疾病的诊断工具。

 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.