Fig. 7: Therapeutic targeting of the CAV1 pathway in HCC.

A Schematic of CAV1 validation in independent HCC cohorts. B Spatial distribution of hepatocyte clusters in HCC. C UMAP visualisation of hepatocytes, coloured by prognosis (recurrence and no recurrence). D CAV1 expression in recurrence-associated hepatocyte clusters. E Bulk RNA-seq analysis of CAV1 expression in HCC patients (n = 55). F Heatmap of CAV1 expression in HCC cell types, clustered by HBV status and recurrence. G CAV1 expression profile in HCC tissues with different etiologies from the TCGA HCC cohort (tumour versus matched normal, n = 50). H Representative histology sections showing CAV1 expression pre- and post-recurrence, with AE1/AE3 (green) as a pan-cytokeratin marker and CAV1 (red) by double-immunofluorescence immunocytochemistry. I RT-qPCR validation of CAV1 knockdown in HCC PDOs using siRNAs. J 3D cell viability assay showing the effect of CAV1 knockdown on HCC PDO growth, with siPLK1 to monitor transfection efficiency. K RT-qPCR of IL-8 and MMP-9 mRNA in CAV1 knockdown PDOs. L 3D cell viability assay assessing lenvatinib sensitivity after CAV1 knockdown in HCC PDOs. M Western blot of signalling pathways following CAV1 knockdown in HCC PDOs. N RT-qPCR of CAV1 mRNA in miR-7 transfected PDOs. O 3D cell viability assay evaluating the effect of miR-7 on HCC PDO growth. P Effect of lenvatinib and BLU9931 combination treatment on HCC PDO growth. Q Effect of sorafenib and BGB324 combination treatment on HCC PDO growth. For siRNA experiments, RNAiMax was used as a lipid control, and non-targeting siRNA served as the lead control. All experiments were performed on three independent days with at least three technical replicates. Error bars represent ± SD. Data were analysed by one-way ANOVA with multiple comparisons ( > 2 groups) or unpaired two-tailed student’s t-test. Significance is denoted as follows: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Lenva = Lenvatinib, SF = Sorafenib.