Short hairpin RNA-directed LDHA silencing using liposomal nanovectors for the effective treatment of human hepatocellular carcinoma

Abstract

Abstract Background Lactate dehydrogenase A (LDHA), which converts pyruvate to lactate, is over-expressed in many malignant tumors. We will analyze the correlation between LDHA expression and clinicopathological features of HCC. We further applied biodegradable polyethyleneimine containing multiple disulfide bonds (SS-PEI) as an effective non-viral vector for the safe intracellular delivery of plasmid LDHA-shRNA-EGFP in vitro and in vivo for the treatment of liver cancer. Methods We first retrospectively analyzed the correlation between the expression of LDHA in liver cancer and the clinical case characteristics of patients and the prognosis of anti-PD1/PD-L1 therapy. Further, I synthesized a nanomaterial SS-PEI/pLDHA-shRNA-EGFP that targets and inhibits the expression of LDHA. Through cytological and animal experiments, it is confirmed that the effectiveness and safety of inhibiting tumor growth. Results Our studies showed that patients with hepatocellular carcinoma (HCC) with high LDHA expression have poorer overall survival and disease-free survival. The expression of PD-L1 in HCC was positively correlated with the expression of Ki-67, PD-L1 and the infiltration of Regulatory T cells, CAF cells in the tumor microenvironment (p < 0.05). The objective response rate of anti-PD1/PD-L1 immunotherapy in HCC patients with low LDHA expression was significantly higher than patients with high LDHA expression(p < 0.05). Transfection experiments showed that the SS-PEI/pLDHA-shRNA-EGFP complexes could be used to transfect various types of HCC cell lines in vitro, inducing reduced expression of LDHA and cell growth inhibition. In addition, treatment of mice with SS-PEI/pLDHA-shRNA-EGFP by tail vein injection significantly inhibited the growth of subcutaneous xenograft tumors. Meanwhile, the complexes revealed relatively low cytotoxicity in vitro and no toxicity was observed in vivo. Conclusions The expression of LDHA in HCC is closely related to tumor proliferation and immune escape. Patients with low LDHA expression of HCC are more sensitive to anti-PD1/PD-L1 immunotherapy. Targeting LDHA in vivo with a specific short hairpin RNA delivered via SS-PEI is a promising therapy for patients with HCC.