Annexin A2 as a Therapeutic Target of Artemisiae Scopariae Herba-derived Quercetin in Preventing Malignant Progression from Primary Sclerosing Cholangitis to Cholangiocarcinoma: An Integrative Study.

ETHNOPHARMACOLOGICAL RELEVANCE: Artemisiae Scopariae Herba (ASH) is traditionally used to treat cholestatic liver diseases and exhibits anti-tumour potential. Cholangiocarcinoma (CCA) and its major risk factor, primary sclerosing cholangitis (PSC), lack effective therapies. AIM OF THE STUDY: In this study, we aimed to elucidate the bioactive constituents and underlying protective mechanisms of ASH in preventing malignant progression from PSC to CCA. MATERIALS AND METHODS: We identified the active constituents of ASH and their targets using multiple databases and mass spectrometry. Integrative transcriptomic analysis of public datasets determined the therapeutic targets. Machine learning and molecular docking predicted key targets, while surface plasmon resonance experimentally confirmed the primary drug-target interaction. Subsequent clinical sample analysis validated these targets. Finally, we assessed the in vitro effects of quercetin on CCA cells and utilised single-cell transcriptomics to characterise target expression within the disease microenvironment. RESULTS: Quercetin emerged as the principal bioactive constituent of ASH, with annexin A2 (ANXA2) as its key target. ANXA2 was overexpressed in CCA and PSC, which correlated with poor patient prognosis. In vitro, quercetin suppressed CCA cell malignant phenotypes and downregulated ANXA2 expression. Single-cell analysis revealed that epithelial cells, functioning as critical communication hubs, primarily overexpressed ANXA2. Functional enrichment analysis implicated pathways regulating the actin cytoskeleton and mediating bacterial infection responses. CONCLUSIONS: Quercetin likely exerts its therapeutic effects by targeting ANXA2 and modulating cholangiocyte pathogenicity. These findings highlight ANXA2 as a promising therapeutic target for treating advanced CCA and halting PSC-driven carcinogenesis.