In vivo production of anti-CD19 CAR-T cells with T cell-targeted engineered exosomes to evaluate cytotoxicity against lymphoma cells
10.12354/j.issn.1000-8179.2025.20250297
- VernacularTitle:靶向T细胞的工程化外泌体在体生成CD19 CAR-T细胞及其对淋巴瘤细胞的杀伤研究
- Author:
Dong TING
1
;
Zhou YING
1
;
Yu BOYU
1
;
Xia XUEJIAO
1
;
Ma YIGE
1
;
Ma YAN
1
;
Gao YANG
1
;
Zhou MENGYING
1
;
Wang CHANGJUN
1
;
Li QIUYI
1
;
Gu CHAOJIANG
1
Author Information
1. 武汉科技大学生命科学与健康学院(武汉市 430065)
- Publication Type:Journal Article
- Keywords:
in vivo generation of CAR-T cells;
CD8-targeted exosome delivery system;
humanized mouse model;
B lymphoma cell line
- From:
Chinese Journal of Clinical Oncology
2025;52(6):279-286
- CountryChina
- Language:Chinese
-
Abstract:
Objective:Chimeric antigen receptor T-cell(CAR-T)immunotherapy has made major breakthroughs in the treatment of blood tu-mors.However,current CAR-T therapies face several limitations:they require autologous cells,involve a lengthy and costly production pro-cess,and use lentiviral transduction that carry risk of insertional carcinogenesis due to random integration.Therefore,there is an urgent need to develop a universal cost-effective cancer immunotherapy method generating CAR-T cells for in vivo cancer immunotherapy.Meth-ods:This study successfully established an exosome-mediated,T-cell targeted delivery system,demonstrating both precise design and func-tional efficacy for biomedical applications.To optimize CAR-T cell generation the transfection dose was adjusted,and the kinetics of CAR-T cell percentage were recorded.The cytotoxicity of the resulting CAR-T cells was evaluated in vitro by calcein-AM release.To test the tumor-killing in vivo of engineered exosomes,human PBMCs were injected into NPG mice via the tail vein to establish humanized mice,followed by intravenous injection of tumor cells to induce cancer.Results:To overcome the limitations of conditional autologous CAR-T cells,we de-veloped a T cell-targeted exosome system capable of specifically targeting human CD3+,CD4+,and CD8+T cells.CAR-T production was dose-dependent,with transfection efficiency reaching upto 97.8%at 106 particles/cell.Both in vitro cytotoxicity assays and in vivo animal experi-ments demonstrated that exosome-incubated CAR-T cells effectively eliminated CD19-positive Raji cells,highlighting their specificity and therapeutic potential in antigen-directed applications.Conclusions:We successfully established a CD8-targeting exosome delivery system for CAR-T cell production capable of transforming CD8+T cells into functional CAR-T cells,which showed significant tumor-killing ability in vitro and in mice.Compared with the traditional lentiviral vector for the preparation of CAR-T cells in vitro,in vivo-reprogrammed CAR-T cells us-ing our CD8-targeted exosome delivery system,with higher transfection efficiency,shorter production period,lower cost,and eliminated the risk of insertion carcinogenesis.This strategy promises to bring a new era of universal CAR-T medicine,which can improve cancer immuno-therapy and may hold promise as a therapeutic platform to treat various diseases.
