白细胞介素-12（IL-12）基因转移增强了采用T细胞治疗的治疗效力。我们先前报告，将癌特异性CD8 T细胞暂时工程成表达IL-12 mRNA，可以提高它们在肿瘤内部输送时的系统治疗功效。在这里，我们将mRNA工程的T细胞混合使用，以表达单链IL-12（scIL-12）或对IL-18结合蛋白（IL-18BP）没有功能性障碍的IL-18诱饵耐药变异体（DRIL18）。重复将混合的mRNA工程T细胞注射到小鼠肿瘤中。经过电穿孔使Pmel-1 T细胞受体（TCR）易位表达scIL-12或DRIL18 mRNA，在局部和远处的黑色素瘤病变中表现出强大的治疗效果。这些效应与T细胞代谢适应性、增强miR-155对免疫抑制靶基因的控制、多种细胞因子的增强表达以及表面蛋白的糖基化谱的改变有关，从而使其粘着E-选择素。在IL-12和DRIL18 mRNA电穿孔下，该肿瘤内淋巴细胞（TILs）和嵌合抗原受体（CAR）T细胞的体外培养重现了这种肿瘤内免疫治疗策略的疗效。版权所有©2023年作者。由Elsevier公司出版。保留所有权利。

Interleukin-12 (IL-12) gene transfer enhances the therapeutic potency of adoptive T cell therapies. We previously reported that transient engineering of tumor-specific CD8 T cells with IL-12 mRNA enhanced their systemic therapeutic efficacy when delivered intratumorally. Here, we mix T cells 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 T cell mixtures are repeatedly injected into mouse tumors. Pmel-1 T cell receptor (TCR)-transgenic T cells electroporated with scIL-12 or DRIL18 mRNAs exert powerful therapeutic effects in local and distant melanoma lesions. These effects are associated with T cell 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) T cells on IL-12 and DRIL18 mRNA electroporation.Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.