Ovarian Cancer Immunotherapy Using PD-L1 siRNA Targeted Delivery from Folic Acid-Functionalized Polyethylenimine: Strategies to Enhance T Cell Killing

Adoptive T cell immunotherapy is a promising treatment strategy for epithelial ovarian cancer (EOC). However, programmed death ligand‐1 (PD‐L1), highly expressed on EOC cells, interacts with programmed death‐1 (PD‐1), expressed on T cells, causing immunosuppression. This study aims to block PD‐1/PD‐L1 interactions by delivering PD‐L1 siRNA, using various folic acid (FA)–functionalized polyethylenimine (PEI) polymers, to SKOV‐3‐Luc EOC cells, and investigate the sensitization of the EOC cells to T cell killing. To enhance siRNA uptake into EOC cells, which over express folate receptors, PEI is modified with FA or PEG–FA so that siRNA is complexed into nanoparticles with folate molecules on the surface. PEI modification with a single functional group lowers the polymer cytotoxicity compared to unmodified PEI. FA‐conjugated polymers increase siRNA uptake into SKOV‐3‐luc cells and decrease unspecific uptake into monocytes. All polymers result in 40% to 50% PD‐L1 protein knockdown. Importantly, SKOV‐3‐Luc cells treated with either PEI–FA or PEI– polyethylene glycol (PEG)–FA/PD‐L1 siRNA complexes are up to twofold more sensitive to T cell killing compared to scrambled siRNA treated controls. These findings are the first to demonstrate that PD‐L1 knockdown in EOC cells, via siRNA/FA‐targeted delivery, are able to sensitize cancer cells to T cell killing. Programmed cell death ligand 1 knockdown on ovarian cancer cells by folic acid (FA) functionalized polyethylenimine (PEI) polymers complexed with siRNA increases T cell functionality and cancer cell killing. FA modified PEI exerts lowered cytotoxicity, increases siRNA uptake in ovarian cancer cells, while decreasing unspecific uptake into phagocytotic mononuclear cells.