MRNAs encoding IL-12 and a decoy-resistant variant of IL-18 synergize to engineer T cells for efficacious intratumoral adoptive immunotherapy

Abstract

Interleukin-12 (IL-12) gene transfer enhances the therapeutic potency of adoptive Tcell therapies. We previously reported that transient engineering of tumor-specific CD8 Tcells with IL-12 mRNA enhanced their systemic therapeutic efficacy when delivered intratumorally. Here, we mix Tcells engineered with mRNAs to express either single-chain IL-12 (scIL-12) or an IL-18 decoy-resistant variant (DRIL18) that is not functionally hampered by IL-18 binding protein (IL-18BP). These mRNA-engineered Tcell mixtures are repeatedly injected into mouse tumors. Pmel-1 Tcell receptor (TCR)-transgenic Tcells electroporated with scIL-12 or DRIL18 mRNAs exert powerful therapeutic effects in local and distant melanoma lesions. These effects are associated with Tcell metabolic fitness, enhanced miR-155 control on immunosuppressive target genes, enhanced expression of various cytokines, and changes in the glycosylation profile of surface proteins, enabling adhesiveness to E-selectin. Efficacy of this intratumoral immunotherapeutic strategy is recapitulated in cultures of tumor-infiltrating lymphocytes (TILs) and chimeric antigen receptor (CAR) Tcells on IL-12 and DRIL18 mRNA electroporation.