Enzyme‐responsive 612‐AYR nanoparticles for targeted inhibition of B4GALT3‐mediated ITGB1 glycosylation in hepatocellular carcinoma

BACKGROUND AND PURPOSE: Aberrant protein glycosylation contributes significantly to hepatocellular carcinoma (HCC) progression. β-1,4-Galactosyltransferase 3 (B4GALT3), an enzyme involved in glycosylation, is overexpressed in HCC and promotes tumour growth by stabilizing integrin β1 (ITGB1). This study aimed to evaluate B4GALT3 as a therapeutic target and develop a precision nanotherapeutic approach for HCC. EXPERIMENTAL APPROACH: Bioinformatics analyses using TCGA-LIHC datasets identified B4GALT3 as a prognostic biomarker in HCC. Functional assays assessed the role of B4GALT3 in proliferation, migration, invasion and ITGB1 glycosylation in HCC cells. Subsequently, enzyme-responsive 612-AYR nanoparticles were engineered to deliver a specific B4GALT3 inhibitor selectively to tumour sites. The nanoparticles' biocompatibility, tumour-targeting capability, retention and therapeutic efficacy were evaluated in both subcutaneous and orthotopic HCC mouse models. KEY RESULTS: B4GALT3 enhanced HCC cell proliferation, migration, and invasion by increasing ITGB1 glycosylation, stabilizing its protein level and activating downstream signalling and cell-cycle pathways. The 612-AYR nanoparticles demonstrated excellent biocompatibility, tumour-specific accumulation and prolonged retention in vivo. Treatment with these nanoparticles markedly suppressed tumour growth and metastasis in both mouse models through effective disruption of the B4GALT3-ITGB1 signalling axis. CONCLUSION AND IMPLICATIONS: Targeting aberrant glycosylation via enzyme-responsive 612-AYR nanoparticles delivering a B4GALT3 inhibitor presents a promising precision therapeutic approach for HCC. This integrated strategy combining glycosylation-targeted molecular biology and advanced nanotechnology offers a novel paradigm for personalized cancer treatment, holding significant therapeutic and diagnostic potential.