Construction of targeting GPR77+CD10+ lipid nanoparticles and validation of targeting capability in vitro and in vivo

The synthesized dual-targeting DOPE (GPR77/CD10) LNP possesses excellent in vitro and in vivo targeting properties, while also having high systemic biosafety and compatibility. • Dual-targeted LNP nanoparticles exhibit excellent stability and drug sustained-release effects. • Dual-targeted LNP nanoparticles have excellent targeting abilities both in vitro and in vivo. • The targeted LNPs possess good biosafety and biocompatibility, providing an important basis for clinical translation. Dual-receptor targeting strategies hold promise for improving nanocarrier specificity in complex tumor microenvironments. Herein, we engineered lipid nanoparticles (LNPs) functionalized with GPR77 and CD10 antibodies to exploit receptor co-expression as a mechanism for enhanced targeting. To rigorously validate this approach, we developed a GPR77 + CD10 + overexpressing CHO cell model, which served as a controlled system to dissect ligand-receptor interactions. The dual-targeting LNPs (DOPE(GPR77/CD10)) exhibited significantly higher cellular uptake in receptor-positive CHO cells compared to single-targeted or non-targeted formulations, demonstrating synergistic binding efficacy. These LNPs also showed excellent drug encapsulation and prolonged circulation. In a CHO xenograft model, dual-targeting LNPs achieved higher tumor accumulation than non-targeted controls, with minimal off-target organ retention. Biosafety assessments confirmed negligible hemolysis and no hepatorenal toxicity. While this study focused on mechanistic validation in a simplified model, our findings establish a generalizable platform for dual-receptor targeting, with potential applications in stromal or tumor cell-specific drug delivery.