新型阳离子类脂材料C1转染肿瘤抗原NY-ESO-1环状RNA刺激T细胞产生IFN-γ的比较研究  

Comparative study of transfection of tumor antigen NY-ESO-1 circRNA with a novel cationic lipid-like material C1 to stimulate IFN-γproduction in T cells

作　　者：周红 马意朋 汪晓娟 刘凤兰 李彬 乔栋娟 夏小俊 任培根[2] 王明军 Zhou Hong;Ma Yipeng;Wang Xiaojuan;Liu Fenglan;Li Bin;Qiao Dongjuan;Xia Xiaojun;Ren Peigen;Wang Mingjun(Department of Research and Development,Shenzhen Innovation Immunotechnology Co.,Ltd.,Shenzhen 518119,China;Center for Energy Metabolism and Reproduction,Shenzhen Institutes of Advanced Technology,Chinese Academy of Sciences,Shenzhen 518055,China;State Key Laboratory of Oncology in South China,Sun Yat-sen University Cancer Center,Guangzhou 510060,China)

机构地区：[1]深圳因诺免疫有限公司研究发展部,深圳518119 [2]中国科学院深圳先进技术研究院能量代谢与生殖研究中心,深圳518055 [3]中山大学肿瘤防治中心,华南肿瘤学国家重点实验室,广州510060

出　　处：《中华微生物学和免疫学杂志》2024年第9期771-777,共7页Chinese Journal of Microbiology and Immunology

基　　金：深圳市科技计划项目(CJGJZD20200617102403009);广东省基础与应用基础研究基金联合基金(2020B1515120018)。

摘　　要：目的利用环状RNA(circular RNA,circRNA)分子表达NY-ESO-1抗原表位,并利用一种新型类脂材料C1制备circRNA肿瘤疫苗,初步评价其在细胞内的转染效率和对T细胞的激活情况。方法体外转录合成表达增强绿色荧光蛋白(enhanced green fluorescent protein,EGFP)和NY-ESO-1抗原表位的mRNA和circRNA,并转染COS7细胞,体外检测目的蛋白的表达情况。利用C1材料和商用转染试剂TransIT-mRNA作为mRNANY-ESO-1和circRNANY-ESO-1的递送系统制备疫苗,比较其转染效率。结果mRNAEGFP、circRNAEGFP体外转染COS7细胞24 h后,荧光显微镜下可见明显的EGFP表达。mRNANY-ESO-1、circRNANY-ESO-1转染COS7-A*02:01细胞后可以刺激靶向NY-ESO-1/HLA-A2的T细胞受体工程化T细胞(T cell receptor-engineered T cells,TCR-T)分泌细胞因子IFN-γ。circRNANY-ESO-1能够更持久地表达目的抗原并刺激靶细胞释放IFN-γ,在体外对细胞株共培养的条件下,C1材料的递送效果较商用转染试剂好。结论相较于线性mRNA,circRNA转染COS7-A*02:01细胞后能够更有效、更持久地激活T细胞,未来可能是一种更适合用于临床治疗肿瘤的分子。类脂材料C1能够有效递送线性mRNA分子和circRNA分子。本研究为circRNA肿瘤疫苗的研究提供了新思路。ObjectiveTo express NY-ESO-1 epitopes using circular RNA(circRNA)and construct circRNA cancer vaccines using a novel lipid-like material C1,and to evaluate the transfection efficiency and T cell activation potential at cellular level.Methods In vitro transcription was used to synthesize mRNA and circRNA expressing EGFP and NY-ESO-1 epitopes.Then,they were transfected into COS7 cells and the expression of target proteins were detected in vitro.Lipid-like material C1 and commercial transfection agent TransIT-mRNA were used as delivery systems for mRNA NY-ESO-1 and circRNA NY-ESO-1,and their delivery efficiency was compared.ResultsThe expression of EGFP was observed under fluorescence microscopy after transfection of mRNA EGFP and circRNA EGFP into COS7 cells for 24 h.The secretion of IFN-γby T cell receptor-engineered T(TCR-T)cells targeting NY-ESO-1/HLA-A2 was stimulated by COS7-A*02:01 cells transfected with mRNA NY-ESO-1 and circRNA NY-ESO-1.Compared with mRNA NY-ESO-1,circRNA NY-ESO-1 was able to express the target antigen and stimulate the target cells to release IFN-γmore persistently.The delivery efficiency of C1 material was better than that of commercial transfection reagents when COS7 cells were transfected in vitro.ConclusionsCompared with the linear mRNA,transfection of COS7-A*02:01 cells with circRNA can lead to more efficient and durable activation of T cells,suggesting that it could be a more suitable candidate for clinical treatment of tumors in the future.The lipid-like material C1 can effectively deliver linear mRNA and circular RNA molecules.This study provides reference for further research on circRNA tumor vaccines.

关 键 词：肿瘤疫苗 NY-ESO-1 circRNA疫苗 类脂材料C1 免疫治疗 

分 类 号：R730.5[医药卫生—肿瘤]