Streamlined and comprehensive circulating tumor exosome profiling by microfluidic arrayed nanoplasmonic sensors and actuators

项目总结 检测血液中的癌症生物标记物，也就是所谓的“液体活检”，原则上可以改善 测量几乎看不见的“微小残留病(MRD)”的准确性。外切体是细胞- 排出的胞外小泡含有表面蛋白质和遗传物质(DNA和RNA) 它们反映了亲代细胞的特征和组成。分析Exosome将会 因此提供了对癌细胞状态的直接洞察。用于癌症诊断 特别是，最近的证据表明，几个微小RNA在 CTE。因此，解锁CTE中的信息财富可能会导致一种范式 换档。然而，目前分析CTE的障碍如下：(1)所有现有技术 需要抽血；(2)涉及复杂的程序；(3)缺乏无标签的大小/计数 分子特异性；(4)提供高度平均的结果，具有来自正常的高背景 外体，从而导致敏感度低。(5)提供“部分”信息：表面抗原 或货运DNA/RNA，但不能两者兼而有之。所有这些都导致了一种简单化的二元结果， 动态范围，不能在高灵敏度的情况下频繁使用。我们建议多管齐下 微流控阵列纳米等离子体传感器和致动器(MANSA)平台上的解决方案：(1) 简化了隔离、集中和分析。(2)通过监测提高敏感度 个体未标记外切体结合事件与动态成像技术的补充 光谱成像。(3)通过分析表面抗原和内部抗原来提高特异性 单个外切体水平的D/RNA生物标志物。(4)使用综合解决方案消除抽血 针头装置。(5)癌细胞系不同样本复杂度的基准性能 提取癌症患者的血液样本。我们的目标是获得一个高分辨率的数字外显体 同时具有多重表面蛋白和货物D/RNA生物标志物图谱，以促进高 动态范围枚举和提升灵敏度。拟议的技术将成为一项成本- 高效、对护理点友好的转换平台，将在早期满足关键需求 癌症和MRD检测，以改善癌症保健结果。这项技术也可以 广泛应用于基于外显子的非癌症疾病诊断和基础生物医学 研究。

PROJECT SUMMARY Detection of cancer biomarkers in the blood, known as “liquid biopsy”, can in principle improve the accuracy of measuring nearly invisible “minimal residual disease (MRD)”. Exosomes are cell- excreted extracellular vesicles that contain surface proteins and genetic materials (DNA and RNA) that reflect the characteristics and make-up of the parental cell. Analyzing exosomes would therefore provide direct insight into the state of the cancerous cell. For cancer diagnostics in particular, recent evidences have shown that several micro-RNAs are differentially expressed in CTE. Therefore, unlocking the wealth of information in CTE can potentially cause a paradigm shift. However, current barriers for profiling CTE are the following: (1) all existing technologies require blood withdrawal; (2) involve sophisticated protocols; (3) label-free sizing/counting lacks molecular specificity; (4) provide highly averaged results with high background from normal exosomes, thus leading to poor sensitivity. (5) provide “partial” information: either surface antigen or cargo DNA/RNA, but not both. All of the above has led to a simplistic binary outcome that lacks dynamic range and cannot be used frequently with high sensitivity. We propose a multi-pronged solution on a microfluidic arrayed nanoplasmonic sensor & actuator (MANSA) platform for: (1) streamlined isolation, concentration, and profiling. (2) improve sensitivity by monitoring individual unlabeled exosome binding events with dynamic imaging technology complemented by spectroscopic imaging. (3) improve specificity by profiling both surface antigen and internal D/RNA biomarkers at single exosome level. (4) eliminate blood withdrawal using an integrated needle device. (5) benchmark performance with various sample complexity from cancer cell line extracts to cancer patient blood samples. Our goal is to obtain a high-resolution, digital exosome map with both multiplex surface protein and cargo D/RNA biomarker profiles to facilitate high dynamic range enumeration and boost sensitivity. The proposed technology will become a cost- effective, point-of-care-friendly, translational platform that will address a critical need in early cancer and MRD detection to improve cancer healthcare outcomes. The technology can also be broadly applied to exosome-based diagnostics of non-cancer diseases and basic biomedical research.