Mechanical characterization of hyaluronic acid-modified cationic liposomes for targeted deliver of ONECUT2 shRNA in hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is a globally significant malignancy with high morbidity and mortality. Anti-tumor targeted drug therapy is a promising therapeutic strategy, but the strategy faces challenges related to delivery efficiency and mechanical interactions within the tumor microenvironment. In our previous study, we found that the transcription factors ONECUT2 (OC2) and CD44 receptor have important roles in HCC progression. We designed high molecular weight hyaluronic acid-modified cationic liposomes (HMW-CL) to take advantage of the binding affinity between hyaluronic acid and CD44 to deliver plasmid DNA (pshOC2) encoding a short hairpin RNA targeting OC2 to HCC cells. The results showed that the prepared HMW-CL had a uniform particle size of 179.5 nm, a moderate zeta potential of 15.8 mV, a high encapsulation efficiency of 86%, which not only protected pshOC2 from degradation but also ensured favorable mechanical stability under physiological shear stresses. Biomechanical characterization revealed that the liposomes maintained structural integrity under simulated blood flow conditions, with minimal deformation and optimal adhesion to CD44-expressing HCC cells. In vitro experiments, HMW-CL/pshOC2 liposomes were characterized by high transfection efficacy, lysosomal escape, and low cytotoxicity. They could efficiently deliver pshOC2 to cells, affecting HCC cell proliferation, migration, invasion, and triggering apoptosis. Biomechanical assays further confirmed that the liposomes altered the mechanical properties of HCC cells, reducing their stiffness and migratory capacity, which are critical factors in tumor progression. In vivo experiments, intravenous injection of HMW-CL/pshOC2 liposomes effectively reduced OC2 expression in HCC tumors and inhibited tumor growth at low toxicity with an inhibition rate of 81.77%. Our study demonstrated that OC2 may be a candidate gene suitable for HCC targeted-therapy, and our HMW-CL/pshOC2 liposomes were prepared based on the hyaluronic acid/CD44 binding strategy, with good stability, high transfection efficacy, and low cytotoxicity. Moreover, their favorable biophysical and biomechanical properties make them a promising delivery system for HCC therapy, with potential applications in modulating the mechanical microenvironment of tumors.