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

项目摘要 检测血液中的癌症生物标志物，被称为“液体活检”，原则上可以改善 测量几乎不可见的“微小残留病（MRD）”的准确性。外来体是细胞- 含有表面蛋白和遗传物质（DNA和RNA）的分泌细胞外囊泡 反映了母体细胞的特征和组成。分析外泌体 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，几种micro-RNA在细胞中的表达差异， 热膨胀系数。因此，释放CTE中的丰富信息可能会导致一种范式， 班然而，目前用于分析CTE的障碍如下：（1）所有现有技术 需要抽血;（2）涉及复杂的方案;（3）缺乏无标记的大小测定/计数 分子特异性;（4）提供高度平均的结果，具有正常 外泌体，从而导致差的灵敏度。(5)提供“部分”信息：表面抗原 或货物DNA/RNA，但不是两者都有。所有这些都导致了一个简单的二元结果， 动态范围，不能频繁使用，灵敏度高。我们提出了一个多管齐下的 微流体阵列纳米等离子体激元传感器和致动器（曼萨）平台上的解决方案，用于：（1） 简化的隔离、浓缩和分析。(2)通过监测提高灵敏度 用动态成像技术补充单个未标记的外泌体结合事件， 光谱成像(3)通过分析表面抗原和内部抗原来提高特异性 单个外泌体水平的D/RNA生物标志物。(4)使用集成的 针装置。(5)具有来自癌细胞系的各种样品复杂性的基准性能 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泌体 用多重表面蛋白和货物D/RNA生物标志物图谱作图，以促进高表达 动态范围枚举和增强灵敏度。这项技术将成为一项成本- 有效、护理点友好的翻译平台，可满足早期 癌症和MRD检测，以改善癌症医疗保健结果。该技术还可 广泛应用于非癌症疾病的基于外泌体的诊断和基础生物医学 research.

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.