Integrated Microfluidic Exosome Profiling for Early Detection of Cancer

描述(由申请人提供)：无症状早期疾病的检测对大多数癌症类型的有效治疗至关重要。例如，当在I期检测到时，卵巢癌患者的5年存活率超过90%，而晚期疾病的存活率为20%。然而，目前大多数癌症的诊断都是在晚期，这突显了对新的生物标记物、策略和技术的迫切需求。由于越来越多的证据表明循环外切体的生物学功能及其在肿瘤发生和发展中的临床意义，探查循环外切体正在成为非侵入性癌症诊断和治疗反应监测的新范式。肿瘤来源的外切体在人类血液中积累，并在一系列选择性的生物分子中得到丰富，包括信号蛋白、酶、肿瘤抗原、miRNAs和mRNAs。某些肿瘤标志物的结构性释放和外切体浓缩为早期癌症诊断提供了独特的机会。尽管外切体具有诊断癌症的潜力，但由于外切体的有效分离、分子分类和综合表征方面的挑战，外切体的生物学和临床价值仍然很大程度上是未知的。在这里，我们建议通过开发一种创新的微流控技术来解决这一主要障碍，该技术能够实现选择性分离、按表面蛋白拓扑学对亚群进行分类，以及外体的原位多路条形码蛋白质图谱，所有这些都在“样品输入-答案”系统中得到简化。该项目有两个具体目标：1)开发一种集成的微流控外切体轮廓分析(MEPA)，用于微升血浆中循环外切体的分子分析；以及2)以卵巢癌为疾病模型，表征和验证MEPA在癌症早期检测中的潜力。这项研究如果成功，将产生一种变革性的技术，可以显著提高肿瘤来源的外切体的分子表征的分析性能，同时克服传统方法中分离、分析吞吐量和样品消耗过程中外切体丢失/损坏的限制。因此，这项技术应该提供一种前所未有的能力来加速外体研究，为非侵入性地探索肿瘤的生物学和开发用于癌症筛查和早期检测的新的可靠生物标志物打开新的机会。

DESCRIPTION (provided by applicant): Detection of asymptomatic early-stage disease is critical for effective treatment of most cancer types. For instance, when detected at Stage I, the 5-year survival rate for ovarian cancer patients is greater than 90%, versus 20% for advanced-stage disease. However, currently most cancers are diagnosed at late stages, which underscore the pressing need of novel biomarkers, strategies and technologies. Probing circulating exosomes is emerging as a new paradigm for non-invasive cancer diagnosis and monitoring of treatment response, because of growing evidences of their biological functions and clinical implications in tumorigenesis and progression. Tumor-derived exosomes accumulate in human blood and are enriched in a selective repertoire of biomolecules, including signaling proteins, enzymes, tumor antigens, miRNAs, and mRNAs. The constitutive release and exosome enrichment of certain tumor markers present distinctive opportunities for early cancer diagnosis. Despite the potential for cancer diagnosis, biology and clinical value of exosomes remain largely unknown, due to the challenges in efficient isolation, molecular classification and comprehensive characterization of exosomes. Here we propose to tackle this major roadblock by developing an innovative microfluidic technology that enables selective isolation, subpopulation classification by surface protein topography, and in situ multiplexed barcode protein profiling of exosomes, all streamlined in a "sample-in-answer-out" system. The project consists of two specific aims: 1) Develop an integrated Microfluidic Exosome Profiling Assay (MEPA) for molecular analysis of circulating exosomes in microliter volumes of plasma; and 2) Characterize and validate MEPA for potential use in early detection of cancer using ovarian cancer as the disease model. This research, if successful, will yield a transformative technology that can substantially improve the analytical performance for molecular characterization of tumor-derived exosomes while overcoming the constraints of exosome loss/damage during isolation, analysis throughput, and sample consumption in conventional protocols. Thus the technology should offer an unprecedented ability to accelerate the exosome research, opening new opportunities to probing the biology of a tumor noninvasively and to developing novel reliable biomarkers for screening and early detection of cancer.