Identifying the Phenotypes of Tumor-Derived Extracellular Vesicles Using Size-Coded Affinity Microbeads.
Identifying the Phenotypes of Tumor-Derived Extracellular Vesicles Using Size-Coded Affinity Microbeads.
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DOI：
10.1021/jacs.2c10042
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发表时间：
2022-12
影响因子：
15
通讯作者：
Jiacheng Wu;Zhun Lin;Zhengyu Zou;Siping Liang;Minhao Wu;Tony Y. Hu;Yuanqing Zhang
中科院分区：
文献类型：
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作者：
Jiacheng Wu;Zhun Lin;Zhengyu Zou;Siping Liang;Minhao Wu;Tony Y. Hu;Yuanqing Zhang
肿瘤源性细胞外囊泡（tEV）生物标志物能够反映癌细胞的表型，在癌症诊断和治疗方面具有很大的潜力。然而，tev表现出高度的异质性，由于其低表达，快速、敏感地鉴定EV生物标志物仍然具有挑战性。光谱重叠也极大地限制了荧光探针对EV生物标志物的多重分析。在此，我们开发了一种高度敏感的tEV表型分析方法，该方法使用带有发夹探针的大小编码微珠，可以结合针对不同tEV生物标志物的适体。我们还设计了一种包含间隔阵列的微流控芯片，该芯片根据微珠的大小将这些微珠隔离在不同的芯片区域，以产生指示不同EV生物标志物水平的特定位置信号。利用原位滚动循环扩增技术（RCA）扩增这些微珠上的EV生物标志物信号。该策略允许荧光光谱同时检测多种tEV表型，而不受光谱重叠的限制。本研究表明，该方法可以快速、同时检测6种不同的tEV表型，灵敏度高。由于传感平台的可编程性，该方法可以快速适应检测被检测生物标志物的不同tEV表型替代。值得注意的是，临床队列研究表明，这一策略可能为癌症患者的精确诊断和个性化治疗提供新的思路。
Tumor-derived extracellular vesicle (tEV) biomarkers can reflect cancer cell phenotypes and have great potential for cancer diagnosis and treatment. However, tEVs display high heterogeneity, and rapid and sensitive identification of EV biomarkers remains challenging due to their low expression. Spectral overlap also significantly limits the multiplex analysis of EV biomarkers by fluorescent probes. Herein, we developed a method for highly sensitive tEV phenotyping that uses size-coded microbeads that carry hairpin probes that can bind to aptamers targeting distinct tEV biomarkers. We also designed a microfluidic chip containing spacer arrays that segregate these microbeads in distinct chip regions according to their size to generate location-specific signals indicating the level of different EV biomarkers. The EV biomarker signal on these microbeads was amplified by in situ rolling cyclic amplification (RCA). This strategy permits the simultaneous detection of multiple tEV phenotypes by fluorescence spectroscopy without the limitations of spectral overlap. This study demonstrates that this tEV phenotyping method can rapidly and simultaneously detect six different tEV phenotypes with high sensitivity. Due to the programmability of the sensing platform, this method can be rapidly adapted to detect different tEV phenotype substitutions of the detected biomarkers. Notably, clinical cohort studies show that this strategy may provide new ideas for the precise diagnosis and personalized treatment of cancer patients.