Unleashing the potential of a low CpG Passer transposon for superior CAR-T cell therapy
BackgroundTo date, the non-viral vector Chimeric Antigen Receptor (CAR) T cell preparation platform, exemplified by transposons, has demonstrated significant potential in tumor immunotherapy and yielded positive results in multiple clinical trials. Nonetheless, non-methylated CpG sequences within pl...
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Frontiers Media S.A.
2025-02-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2025.1541653/full |
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| author | Jianyao Zeng Yan Sun Yan Sun Yuan Fang Xiaodie Wang Qian Huang Pingjing Zhang Meiqi Shao Pei Wang Jingbo Cheng Meng Di Tao Liu Qijun Qian Qijun Qian Qijun Qian |
| author_facet | Jianyao Zeng Yan Sun Yan Sun Yuan Fang Xiaodie Wang Qian Huang Pingjing Zhang Meiqi Shao Pei Wang Jingbo Cheng Meng Di Tao Liu Qijun Qian Qijun Qian Qijun Qian |
| author_sort | Jianyao Zeng |
| collection | DOAJ |
| description | BackgroundTo date, the non-viral vector Chimeric Antigen Receptor (CAR) T cell preparation platform, exemplified by transposons, has demonstrated significant potential in tumor immunotherapy and yielded positive results in multiple clinical trials. Nonetheless, non-methylated CpG sequences within plasmid DNA can elicit an inflammatory response via Toll-like receptor 9 (TLR9) during CAR-T cell preparation, adversely affecting transgene expression. Additionally, de novo DNA methylation programs promote T cell exhaustion, which poses a significant limitation for CAR-T cell therapy applications.MethodsHigh-throughput liquid protein chip and CBA analyses were utilized to determine the expression levels of inflammatory factors. Flow cytometry and luciferase reporter assays were employed for mutation screening. BALB/c mice and M-NSG mice were used to evaluate the inflammatory response and efficacy of LCG CAR-T in vivo, with TIL grouping detected via immunohistochemistry.ResultsIn this study, we modified the newly discovered Passer (JL) transposon to construct a low-CpG content transposon for CAR-T cell (LCG CAR-T cell) preparation. In vitro experiments demonstrated that LCG CAR-T cells prepared using this new transposon exhibited stronger cytotoxicity. In animal models, LCG CAR-T cells significantly inhibited tumor growth and increased the populations of CD4+CAR-T cells and tumor-infiltrating lymphocytes. Furthermore, LCG CAR-T cells modulated pro-inflammatory cytokine release, thereby reducing in vivo inflammatory responses and surpassing the effects observed with unmodified CAR-T cells.ConclusionsCollectively, our results demonstrate the high safety and efficacy of non-viral, low CpG Passer transposon CAR-T cells, offering new avenues for improving CAR-T cell efficacy while minimizing in vivo inflammation. |
| format | Article |
| id | doaj-art-a2404fb00c5340ab9557da67d2675471 |
| institution | Kabale University |
| issn | 1664-3224 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj-art-a2404fb00c5340ab9557da67d26754712025-02-06T12:09:51ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-02-011610.3389/fimmu.2025.15416531541653Unleashing the potential of a low CpG Passer transposon for superior CAR-T cell therapyJianyao Zeng0Yan Sun1Yan Sun2Yuan Fang3Xiaodie Wang4Qian Huang5Pingjing Zhang6Meiqi Shao7Pei Wang8Jingbo Cheng9Meng Di10Tao Liu11Qijun Qian12Qijun Qian13Qijun Qian14School of Medicine, Shanghai University, Shanghai, ChinaSchool of Medicine, Shanghai University, Shanghai, ChinaInnovative Drugs Business Group, Shanghai Cell Therapy Group, Shanghai, ChinaInnovative Drugs Business Group, Shanghai Cell Therapy Group, Shanghai, ChinaSchool of Medicine, Shanghai University, Shanghai, ChinaInnovative Drugs Business Group, Shanghai Cell Therapy Group, Shanghai, ChinaInnovative Drugs Business Group, Shanghai Cell Therapy Group, Shanghai, ChinaInnovative Drugs Business Group, Shanghai Cell Therapy Group, Shanghai, ChinaInnovative Drugs Business Group, Shanghai Cell Therapy Group, Shanghai, ChinaInnovative Drugs Business Group, Shanghai Cell Therapy Group, Shanghai, ChinaSchool of Medicine, Shanghai University, Shanghai, ChinaInnovative Drugs Business Group, Shanghai Cell Therapy Group, Shanghai, ChinaSchool of Medicine, Shanghai University, Shanghai, ChinaInnovative Drugs Business Group, Shanghai Cell Therapy Group, Shanghai, ChinaShanghai Mengchao Cancer Hospital, Shanghai University, Shanghai, ChinaBackgroundTo date, the non-viral vector Chimeric Antigen Receptor (CAR) T cell preparation platform, exemplified by transposons, has demonstrated significant potential in tumor immunotherapy and yielded positive results in multiple clinical trials. Nonetheless, non-methylated CpG sequences within plasmid DNA can elicit an inflammatory response via Toll-like receptor 9 (TLR9) during CAR-T cell preparation, adversely affecting transgene expression. Additionally, de novo DNA methylation programs promote T cell exhaustion, which poses a significant limitation for CAR-T cell therapy applications.MethodsHigh-throughput liquid protein chip and CBA analyses were utilized to determine the expression levels of inflammatory factors. Flow cytometry and luciferase reporter assays were employed for mutation screening. BALB/c mice and M-NSG mice were used to evaluate the inflammatory response and efficacy of LCG CAR-T in vivo, with TIL grouping detected via immunohistochemistry.ResultsIn this study, we modified the newly discovered Passer (JL) transposon to construct a low-CpG content transposon for CAR-T cell (LCG CAR-T cell) preparation. In vitro experiments demonstrated that LCG CAR-T cells prepared using this new transposon exhibited stronger cytotoxicity. In animal models, LCG CAR-T cells significantly inhibited tumor growth and increased the populations of CD4+CAR-T cells and tumor-infiltrating lymphocytes. Furthermore, LCG CAR-T cells modulated pro-inflammatory cytokine release, thereby reducing in vivo inflammatory responses and surpassing the effects observed with unmodified CAR-T cells.ConclusionsCollectively, our results demonstrate the high safety and efficacy of non-viral, low CpG Passer transposon CAR-T cells, offering new avenues for improving CAR-T cell efficacy while minimizing in vivo inflammation.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1541653/fullmethylationCpG motifchimeric antigen receptor T celltransposoninflammation |
| spellingShingle | Jianyao Zeng Yan Sun Yan Sun Yuan Fang Xiaodie Wang Qian Huang Pingjing Zhang Meiqi Shao Pei Wang Jingbo Cheng Meng Di Tao Liu Qijun Qian Qijun Qian Qijun Qian Unleashing the potential of a low CpG Passer transposon for superior CAR-T cell therapy Frontiers in Immunology methylation CpG motif chimeric antigen receptor T cell transposon inflammation |
| title | Unleashing the potential of a low CpG Passer transposon for superior CAR-T cell therapy |
| title_full | Unleashing the potential of a low CpG Passer transposon for superior CAR-T cell therapy |
| title_fullStr | Unleashing the potential of a low CpG Passer transposon for superior CAR-T cell therapy |
| title_full_unstemmed | Unleashing the potential of a low CpG Passer transposon for superior CAR-T cell therapy |
| title_short | Unleashing the potential of a low CpG Passer transposon for superior CAR-T cell therapy |
| title_sort | unleashing the potential of a low cpg passer transposon for superior car t cell therapy |
| topic | methylation CpG motif chimeric antigen receptor T cell transposon inflammation |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2025.1541653/full |
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