The Translational Research Institute for Neurological Disorders & Interdisciplinary Research Center of Neuromedicine and Chemical Biology of Wannan Medical College, Department of Neurosurgery, Department of Intensive Care Medicine, the First Affiliated Hospital of Wannan Medical College (Yijishan Hospital of Wannan Medical College), Wuhu, China.
Zhongda Hospital, School of Medicine & School of Biological Sciences and Medical Engineering, Advanced Institute for Life and Health & Interdisciplinary Innovation Institute for Medicine and Engineering, Southeast University, Nanjing, China.
Nat Cancer. 2024 Jan;5(1):30-46. doi: 10.1038/s43018-023-00650-8. Epub 2023 Oct 16.
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 hGSDMD-F1L 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-GSDMD-F1L circRNA-LNPs (GSDMD 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, GSDMD 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.
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性 IVT mRNA，它利用人鼻病毒 2 型（HRV2）内部核糖体进入位点（IRES）在 EIF4G2 和 PTBP1 高表达的癌细胞中选择性触发翻译。溶瘤效应由带有靶向线粒体内膜心磷脂的长 hGSDMD-F1L 突变体 mRNA 序列提供，该序列触发线粒体自噬。利用置换内含子-外显子（PIE）剪接环化策略和脂质纳米颗粒（LNP）包封降低了 mRNA 的免疫原性，并能够在体内递送至真核细胞。工程 HRV2 IRES-GSDMD-F1L circRNA-LNP（GSDMD circRNA）成功抑制了 EIF4G2/PTBP1 泛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 异常的自发肿瘤模型中，GSDMD circRNA 显著预防了胰腺、肺和结肠腺癌的发生，提高了存活率并诱导了持续的 KRAS G12D 肿瘤抗原特异性细胞毒性 T 淋巴细胞反应。
