Nano-plasmonic technology for high-throughput single exosome analyses

细胞外囊泡（EV）为癌症诊断和治疗监测提供了新的机会， 循环生物标志物。这些细胞衍生的膜结合囊泡大量存在于生物体液中 并携带细胞特异性的货物，如脂质，蛋白质和遗传物质，可以作为一种 微创手段来探测肿瘤的分子状态。然而，EV分析提出了独特的 由于EV的纳米尺寸、异质性和存在于广泛的生物背景中，因此存在技术挑战。 我们之前已经开发了纳米等离子体传感平台，可以快速灵敏地检测肿瘤 直接来自临床样本的EV。利用我们以前的原型经验， 应用是为了进一步推进尖端的纳米等离子体技术， 以单一EV分辨率进行分析。具体来说，我们将1）适应金纳米棒结构，提供更高的 灵敏度低至单个分子水平，和2）集成等离子体增强荧光检测， 在单个EV中进行多重EV蛋白和RNA分析。我们假设，这种方法将更加 在分析EV蛋白和mRNA标志物方面比目前可能的更敏感和全面。使用 卵巢癌作为一个模型系统，我们将评估1）高通量单EV的系统实现 检测; 2）可靠地检测临床样品中的肿瘤来源的EV;和3）多重EV标志物 筛选肿瘤来源的EV。如果成功，开发的平台将提供一个更容易使用的工具， 显著加速EV分析作为临床癌症护理常规筛查试验的临床采用 设置.

Extracellular vesicles (EVs) present new opportunities for cancer diagnoses and treatment monitoring as circulating biomarkers. These cell-derived membrane-bound vesicles are abundantly present in biological fluids and carry cell-specific cargos, such as lipids, proteins and genetic materials, which can be harnessed as a minimally invasive means to probe the molecular status of tumors. EV analyses, however, poses unique technical challenges due to EVs' nanometer size, heterogeneity, and presence in vast biological background. We have previously developed nanoplasmonic sensing platforms that can rapidly and sensitively detect tumor EVs directly from clinical samples. Drawing upon our previous experiences with the prototypes, the goal of this application is to further advance the cutting-edge nanoplasmonics technology for comprehensive molecular analyses at single EV resolution. Specifically, we will 1) adapt gold nanorod structures that provides higher sensitivity down to a single molecular level and 2) integrate plasmon-enhanced fluorescence detection for multiplexed EV protein and RNA analyses in single EVs. We hypothesize that the approach will be more sensitive and comprehensive in profiling EV protein and mRNA markers than is currently possible. Using an ovarian cancer as a model system, we will evaluate 1) system implementation for high-throughput single EV detection; 2) reliable detection of tumor-derived EVs in clinical samples; and 3) multiplexed EV marker screening on tumor-derived EVs. If successful, the developed platform would render a more accessible tool to significantly accelerate the clinical adoption of EV analyses as routine screening tests for cancer care in clinical settings.