Single Circulating Vesicle Analysis for Early Cancer Detection

循环细胞外囊泡（EV）的分析在过去几年中取得了进展，但挑战 以在临床环境中可重复地分析样品。如果更灵敏和可重复的方法可以 一旦建立，它们将在早期癌症检测、测量患者 对药物的反应，了解肿瘤的异质性和区分不同的疾病。问题是 在胰腺癌（PDAC）中，缺乏早期检测方法， 测量一直是一个问题，缺乏对囊泡异质性的理解。此外，委员会认为， 大多数现有的技术依赖于本体测量，其本质上不太敏感， 利益被稀释（Yang等人，2017，Sci Transl Med，9）。这项提案的目的是探索一种新颖的单一 EV分析（SEA）方法（Lee et al，2018，ACSnano，出版中）揭示了肿瘤EV（TEV）生物标志物 在PDAC中。具体而言，我们建议i）将该方法与芯片上微流体集成，以改善微流体的性能。 样品处理，同时添加MAb-DNA条形码化步骤用于信号扩增和多重化（目的1），和ii） 扩展并测试临床样本中TEV的即时分析方法（目标2）。提议的单身 囊泡方法有可能改变PDAC癌症研究和临床实践。它能让我们发现 临床标本中TEV的组成，在早期检测（稀有蛋白质分析）方面具有深远的应用 以及不同类型癌症的治疗评估。

Analysis of circulating extracellular vesicles (EV) have advanced over the last several years but challenges remain to reproducibly analyze samples in a clinical setting. If more sensitive and reproducible methods could be established, they would have far reaching applications in early cancer detection, measuring patient response to drugs, understanding tumor heterogeneity and discriminating different diseases. The problem is potentiated in pancreatic cancer (PDAC), where early detection methods are lacking, reproducibility of measurements has been a problem and an understanding of vesicle heterogeneity is missing. Furthermore, most existing technologies rely on bulk measurements which are inherently less sensitive since markers of interest are diluted (Yang et al., 2017, Sci Transl Med, 9). The goal of this proposal is to explore a novel single EV analysis (SEA) method (Lee et al, 2018, ACSnano, in press) to shed light on tumor EV (TEV) biomarker composition in PDAC. Specifically, we propose to i) integrate the method with on-chip microfluidics for improved sample handling while adding a MAb-DNA barcoding step for signal amplification and multiplexing (aim 1) and ii) expand and test the approach for point-of-care analyses of TEV in clinical samples (aim 2). The proposed single vesicle method has the potential to transform PDAC cancer research and clinical practice. It will allow us to discover the make-up of TEV in clinical specimen with far reaching applications for early detection (analysis of rare proteins) and treatment t evaluation in different types of cancers.