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于大多数癌症类型的有效治疗至关重要。例如，在第一阶段检测到卵巢癌时，卵巢癌患者的5年生存率大于90%，而晚期患者的5年生存率为20%。然而，目前大多数癌症是在晚期诊断出来的，这强调了对新的生物标志物、策略和技术的迫切需要。由于越来越多的证据表明循环外泌体在肿瘤发生和发展中的生物学功能和临床意义，探测循环外泌体正成为一种非侵入性癌症诊断和治疗反应监测的新范式。肿瘤来源的外泌体在人血液中积累，并富集有选择性的生物分子，包括信号蛋白、酶、肿瘤抗原、mirna和mrna。某些肿瘤标志物的组成释放和外泌体富集为早期癌症诊断提供了独特的机会。尽管外泌体具有癌症诊断的潜力，但由于外泌体的有效分离、分子分类和全面表征方面的挑战，其生物学和临床价值在很大程度上仍然未知。在这里，我们建议通过开发一种创新的微流体技术来解决这一主要障碍，该技术可以实现外泌体的选择性分离，通过表面蛋白质形貌进行亚群分类，以及外泌体的原位多路条形码蛋白质分析，所有这些都在“样品-答案-输出”系统中简化。该项目包括两个具体目标：1)开发一种集成的微流体外泌体分析方法（MEPA），用于微升血浆中循环外泌体的分子分析；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在应用。这项研究如果成功，将产生一种变革性的技术，可以大大提高肿瘤来源外泌体的分子特征分析性能，同时克服传统方案中外泌体在分离、分析吞吐量和样品消耗过程中的损失/损伤的限制。因此，该技术将提供前所未有的加速外泌体研究的能力，为非侵入性探索肿瘤生物学以及开发用于筛查和早期检测癌症的新型可靠生物标志物提供新的机会。

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.