体外转录 (IVT) mRNA 已成为生产核酸药物的快速方法。在这里，我们构建了一种溶瘤 IVT mRNA，它利用人鼻病毒 2 型 (HRV2) 内部核糖体进入位点 (IRES) 选择性触发高表达 EIF4G2 和 PTBP1 的癌细胞中的翻译。溶瘤作用是由长 hGSDMDc .825 T>A/c.884 A>G-F1LCT 突变体 mRNA 序列提供的，其线粒体内膜心磷脂靶向触发线粒体自噬的毒性。利用重排内含子-外显子 (PIE) 剪接环化策略和脂质纳米颗粒 (LNP) 封装降低了 mRNA 的免疫原性，并能够在体内递送至真核细胞。工程化 HRV2 IRESs-GSDMDp.D275E/E295G-F1LCT circRNA-LNPs (GSDMDENG circRNA) 成功抑制 EIF4G2 /PTBP1 全腺癌异种移植物的生长。重要的是，在由KRAS G12D突变引起的EIF4G2和PTBP1异常的自发性肿瘤模型中，GSDMDENG circRNA显着预防胰腺癌、肺腺癌和结肠腺癌的发生，提高存活率并诱导持续的KRAS G12D肿瘤抗原特异性细胞毒性T淋巴细胞应答。 © 2023。作者获得 Springer Nature America, Inc. 的独家许可。

In vitro-transcribed (IVT) mRNA has arisen as a rapid method for the production of nucleic acid drugs. Here, we have constructed an oncolytic IVT mRNA that utilizes human rhinovirus type 2 (HRV2) internal ribosomal entry sites (IRESs) to selectively trigger translation in cancer cells with high expression of EIF4G2 and PTBP1. The oncolytic effect was provided by a long hGSDMDc .825 T>A/c.884 A>G-F1LCT mutant mRNA sequence with mitochondrial inner membrane cardiolipin targeting toxicity that triggers mitophagy. Utilizing the permuted intron-exon (PIE) splicing circularization strategy and lipid nanoparticle (LNP) encapsulation reduced immunogenicity of the mRNA and enabled delivery to eukaryotic cells in vivo. Engineered HRV2 IRESs-GSDMDp.D275E/E295G-F1LCT circRNA-LNPs (GSDMDENG circRNA) successfully inhibited EIF4G2+/PTBP1+ pan-adenocarcinoma xenografts growth. Importantly, in a spontaneous tumor model with abnormal EIF4G2 and PTBP1 caused by KRAS G12D mutation, GSDMDENG circRNA significantly prevented the occurrence of pancreatic, lung and colon adenocarcinoma, improved the survival rate and induced persistent KRAS G12D tumor antigen-specific cytotoxic T lymphocyte responses.© 2023. The Author(s), under exclusive licence to Springer Nature America, Inc.