Key Laboratory of Micro-Nanoscale Bioanalysis and Drug Screening of Guangxi Higher Education, Guangxi Beibu Gulf Marine Biomedicine Precision Development and High-Value Utilization Engineering Research Center, National Center for International Research of Bio-targeting Theranostics, School of Pharmacy, Guangxi Medical University, Nanning, 530021, China.
Department of Clinical Laboratory, The Affiliated Tumor Hospital of Guangxi Medical University, Nanning, 530021, China.
Biosens Bioelectron. 2022 Nov 15;216:114636. doi: 10.1016/j.bios.2022.114636. Epub 2022 Aug 14.
Simultaneous detection of specific exosomal surface proteins and inner microRNAs are hampered by their heterogeneity, low abundance and spatial segregation in nanovesicles. Here, we design a dual-cycling nanoprobe (DCNP) to enable single-step simultaneous quantitation of cancerous exosomal surface programmed death-ligand 1 (PD-L1) (ExoPD-L1) and miRNA-21 (ExomiR-21) directly in exosome lysates, without resorting to either RNA extraction or time-consuming transmembrane penetration. In this design, DNA molecular machine-based dual-recognition probes co-assemble onto gold nanoparticle surface for engineering 'silent' DCNPs, which enable signal-amplified synchronous response to dual-targets as activated by ExomiR-21 and ExoPD-L1 within 20 min. Benefiting from cycling amplification of the molecular machine, DCNPs sensor achieves detection limits of tumor exosomes, ExoPD-L1 and ExomiR-21 down to 10 particles/μL, 0.17 pg/mL and 66 fM, respectively. Such a sensitive dual-response strategy allows simultaneous tracking the dynamic changes of ExoPD-L1 and ExomiR-21 expression regulated by signaling molecules or therapeutics. This approach further detects circulating ExoPD-L1 and ExomiR-21 in human plasma to differentiate breast cancer patients from healthy individuals with high accuracy, showing great potential of DCNPs for simultaneous profiling exosomal surface and inside biomarkers, and for clinical precision diagnosis.
同时检测特定的外泌体表面蛋白和内 microRNAs 受到其异质性、低丰度和在纳米囊泡中的空间隔离的阻碍。在这里，我们设计了一种双循环纳米探针 (DCNP)，可以在单个步骤中直接在外泌体裂解物中定量检测癌外泌体表面程序性死亡配体 1 (PD-L1) (ExoPD-L1) 和 microRNA-21 (ExomiR-21)，而无需进行 RNA 提取或耗时的跨膜渗透。在这种设计中，基于 DNA 分子机器的双重识别探针共同组装到金纳米粒子表面，用于构建“沉默”的 DCNP，其能够通过 ExomiR-21 和 ExoPD-L1 在 20 分钟内激活的双重靶标进行信号放大的同步响应。受益于分子机器的循环扩增，DCNP 传感器实现了对肿瘤外泌体、ExoPD-L1 和 ExomiR-21 的检测限低至 10 个/μL、0.17 pg/mL 和 66 fM。这种敏感的双重响应策略允许同时跟踪信号分子或治疗剂调节的 ExoPD-L1 和 ExomiR-21 表达的动态变化。该方法进一步检测人血浆中的循环 ExoPD-L1 和 ExomiR-21，以高精度区分乳腺癌患者和健康个体，表明 DCNP 在外泌体表面和内部生物标志物的同时分析以及临床精确诊断方面具有巨大潜力。
