PO.CL01.19 · 临床研究
通过整合纳米等离子体传感与多重富集技术设计用于早期PDAC检测的新一代EV诊断方法
Engineering next-generation EV diagnostics by integrating nanoplasmonic sensing and multiplex enrichment for early PDAC detection
该海报暂无可下载的资料
AACR 官方页面
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：胰腺导管腺癌（PDAC）通常在晚期才被诊断，此时可治愈的选择已十分有限。循环细胞外囊泡（EVs）为肿瘤生物学提供了一个微创窗口，但现有的EV检测方法速度慢、耗样量大，且与要求速度、可重复性和最少样本量的临床工作流程不兼容。
方法：为应对这些挑战，我们建立了一套序贯式EV液体活检流程，整合了三个互补的纳米工程平台用于PDAC检测。首先，纳米等离子体增强散射（nPES）检测直接从微升级血浆中捕获EVs，并通过双抗体-纳米颗粒偶联对肿瘤富集的EphA2阳性EVs进行定量，同时能够发现与PDAC相关的其他EV表面标志物。其次，我们引入了FLARE（涨落增强的同步标记与快速富集），其采用低频振动膜和一种模拟过氧化物酶的纳米酶，在单次45分钟操作中同步富集、标记和洗涤EVs，无需多步离心并提升了分析性能。第三，我们将等离子体平台改进为一种多重荧光检测方法，在原位组装等离子体基底以增强来自未处理血浆的EV表面蛋白发射信号，从而能够在一次运行中同时分析多个PDAC相关标志物。
结果：EphA2-EV nPES检测能够对小体积动物样本和患者血浆中的肿瘤来源EVs进行高灵敏度定量，能够将PDAC与胰腺炎及健康供者区分开，并揭示治疗前后EV的动态变化。FLARE的EV回收率比传统方法高十倍以上，同时保持高标记效率，其放大的比色输出可实现基于智能手机的多种EV生物标志物定量，以0.95的曲线下面积区分早期PDAC。多重等离子体增强荧光检测进一步提高了稀有EV蛋白检测的信噪比，无需事先分离EV即可从极少量血浆中实现多标志物读出，有助于个体化预后。
结论：通过结合等离子体散射、振动辅助纳米酶富集和等离子体增强荧光，这套以EV为核心的流程建立了一个快速、低成本且节省样本的PDAC液体活检框架。这些平台共同支持可规模化的早期检测、纵向治疗监测，以及跨临床和临床前环境的大队列转化研究。
查看英文原文 English abstract
Background: Pancreatic ductal adenocarcinoma (PDAC) is typically diagnosed at an advanced stage, when curative options are limited. Circulating extracellular vesicles (EVs) provide a minimally invasive window into tumor biology, yet existing EV assays are slow, sample-intensive, and incompatible with clinical workflows requiring speed, reproducibility, and minimal sample input.
Methods: To confront these challenges, we established a sequential EV liquid-biopsy pipeline that integrates three complementary nanoengineered platforms for PDAC detection. First, a nanoplasmon-enhanced scattering (nPES) assay directly captures EVs from microliter plasma and quantifies tumor-enriched EphA2-positive EVs through dual antibody-nanoparticle coupling, simultaneously enabling the discovery of additional EV surface markers associated with PDAC. Second, we introduce FLARE (Fluctuation-enhanced simultaneous Labeling And Rapid Enrichment), which employs a low-frequency vibrating membrane and a peroxidase-mimicking nanozyme to synchronously enrich, label, and wash EVs in a single 45-minute operation, eliminating the need for multistep centrifugation and improving analytical performance. Third, we refined our plasmonic platform into a multiplex fluorescence assay that assembles plasmonic substrates in situ to enhance EV surface protein emission from unprocessed plasma, allowing simultaneous profiling of multiple PDAC-associated markers within one run.
Results: The EphA2-EV nPES assay enabled highly sensitive quantification of tumor-derived EVs in both small-volume animal samples and patient plasma, distinguished PDAC from pancreatitis and healthy donors, and revealed dynamic EV changes before and after therapy. FLARE achieved more than ten-fold higher EV recovery than conventional methods while maintaining high labeling efficiency, and its amplified colorimetric output enabled smartphone-based quantification of multiple EV biomarkers, discriminating early-stage PDAC with an area under the curve of 0.95. The multiplex plasmon-enhanced fluorescence assay further improved signal-to-noise ratios for rare EV protein detection, enabling multi-marker readout from minimal plasma without prior EV isolation, facilitating personalized prognosis.
Conclusions: By combining plasmonic scattering, vibration-assisted nanozyme enrichment, and plasmon-enhanced fluorescence, this EV-centered pipeline establishes a rapid, low-cost, and sample-sparing framework for PDAC liquid biopsy. Together, these platforms support scalable early detection, longitudinal treatment monitoring, and large-cohort translational studies across both clinical and preclinical settings.
利益披露 Disclosure
T. Hu, None.