Quantification of Fascin-1-Positive Extracellular Vesicles by Nanoflow Cytometry for Early Detection of Hepatocellular Carcinoma in Liquid Biopsy

<b>Background:</b> Hepatocellular carcinoma (HCC) lacks effective early diagnostic biomarkers. Identifying extracellular vesicles (EVs)-associated biomarkers for early HCC detection in HCC progression is precise and critical. <b>Methods:</b> Databases, tissue microarrays (TMA) were used to identify Fascin-1 as a candidate target for HCC, EVs isolated from tumoral and adjacent tissues. The tumor cells were transfected with lentivirus to obtain the engineered EVs. <b>Results:</b> Fascin-1-enriched EVs promoted HCC cell migration and induced cytoskeleton reorganization <i>in vitro</i>. Importantly, quantifying Fascin-1-positive EVs (Fascin-1⁺EVs) in patient plasma by nanoflow cytometry (nFCM) demonstrated high diagnostic performance (AUC=0.8925, 95%CI: 0.7959-0.9891) for discriminating advanced stage from early stage of HCC patients and healthy individuals. <b>Conclusion:</b> Our findings highlight the potential of Fascin-1⁺EVs as a novel non-invasive biomarker for early HCC detection, and quantitative Fascin-1⁺EV analysis by nanoflow cytometry provides a promising approach for HCC diagnosis.