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<p>Extracellular vesicles (EVs) mediate tumor–host communication and represent a promising liquid&#xa0;biopsy source for metastasis risk assessment, yet quantitative detection of low-abundance, epitope-defined EV subpopulations in plasma remains technically challenging. Here, we establish a nanoscale flow cytometry workflow on the CytoFLEX platform for sensitive single-EV phenotyping by optimizing violet side scatter (VSSC) triggering, defining an acquisition window that minimizes coincidence or “swarm” effects, and applying fluorescence-based analysis with stringent background controls. Using this framework, we quantified EV-associated tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) at the single particle level, with good inter-assay reproducibility (CV ~ 11–13%), and resolved low-abundance TRAIL⁺ EVs at approximately 1% abundance within total EV events. Due to the low abundance of EV-associated TRAIL in pancreatic ductal adenocarcinoma (PDAC) plasma, ELISA lacked sufficient analytical sensitivity to accurately reflect EV-associated TRAIL levels, whereas flow-based enumeration preserved quantitative resolution. Clinically, plasma EV-associated TRAIL was significantly elevated in PDAC patients with liver metastasis and demonstrated predictive utility for postoperative liver metastatic recurrence (AUC = 0.766). These results support nanoscale flow cytometry as a robust platform for plasma EV biomarker profiling and identify EV-associated TRAIL as an informative indicator of liver metastatic risk in PDAC.</p>

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Detection of plasma EV-associated TRAIL by nanoscale flow cytometry for liver metastasis prediction in PDAC

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年3月4日

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  • Chun-Xiang Huang,
  • Jia-Hong Jian,
  • Jun-Sheng Hao,
  • Zi-Wen Zhou,
  • Zhuo-Qun Li,
  • Dong-Ming Kuang,
  • Cai-Yuan Wu

摘要

Extracellular vesicles (EVs) mediate tumor–host communication and represent a promising liquid biopsy source for metastasis risk assessment, yet quantitative detection of low-abundance, epitope-defined EV subpopulations in plasma remains technically challenging. Here, we establish a nanoscale flow cytometry workflow on the CytoFLEX platform for sensitive single-EV phenotyping by optimizing violet side scatter (VSSC) triggering, defining an acquisition window that minimizes coincidence or “swarm” effects, and applying fluorescence-based analysis with stringent background controls. Using this framework, we quantified EV-associated tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) at the single particle level, with good inter-assay reproducibility (CV ~ 11–13%), and resolved low-abundance TRAIL⁺ EVs at approximately 1% abundance within total EV events. Due to the low abundance of EV-associated TRAIL in pancreatic ductal adenocarcinoma (PDAC) plasma, ELISA lacked sufficient analytical sensitivity to accurately reflect EV-associated TRAIL levels, whereas flow-based enumeration preserved quantitative resolution. Clinically, plasma EV-associated TRAIL was significantly elevated in PDAC patients with liver metastasis and demonstrated predictive utility for postoperative liver metastatic recurrence (AUC = 0.766). These results support nanoscale flow cytometry as a robust platform for plasma EV biomarker profiling and identify EV-associated TRAIL as an informative indicator of liver metastatic risk in PDAC.