Differences in the phenotypes and transcriptomic signatures of chimeric antigen receptor T lymphocytes manufactured via electroporation or lentiviral transfection

Frontiers in Immunology

Immunology

Zhang Zhang¹  Jintao Zou¹  Wei Chen¹  Xiaopeng Zhang¹  Xuan Hu¹  Anna Niu¹  Jing Wang¹  Lingyu Su²  Xing Lu²  Wei Zhang³ 

[1] Beijing Institute of Biotechnology, Beijing, China;Beijing Institute of Biotechnology, Beijing, China;Nanhu Laboratory, Jiaxing, Zhejiang, China;Nanhu Laboratory, Jiaxing, Zhejiang, China;

关键词: chimeric antigen receptor T lymphocyte;    piggyBac transposon;    lentiviral transfection;    phenotype;    transcriptomic signature;   

DOI  :  10.3389/fimmu.2023.1068625

 received in 2022-10-13, accepted in 2023-04-14,  发布年份 2023

来源: Frontiers

【 摘 要 】

Chimeric antigen receptor (CAR)-T cell therapy is an innovative treatment for CD19-expressing lymphomas. CAR-T cells are primarily manufactured via lentivirus transfection or transposon electroporation. While anti-tumor efficacy comparisons between the two methods have been conducted, there is a current dearth of studies investigating the phenotypes and transcriptome alterations induced in T cells by the two distinct manufacturing methods. Here, we established CAR-T signatures using fluorescent imaging, flow cytometry, and RNA-sequencing. A small fraction of CAR-T cells that were produced using the PiggyBac transposon (PB CAR-T cells) exhibited much higher expression of CAR than those produced using a lentivirus (Lenti CAR-T cells). PB and Lenti CAR-T cells contained more cytotoxic T cell subsets than control T cells, and Lenti CAR-T cells presented a more pronounced memory phenotype. RNA-sequencing further revealed vast disparities between the two CAR-T cell groups, with PB CAR-T cells exhibiting greater upregulation of cytokines, chemokines, and their receptors. Intriguingly, PB CAR-T cells singularly expressed IL-9 and fewer cytokine release syndrome-associated cytokines when activated by target cells. In addition, PB CAR-T cells exerted faster in vitro cytotoxicity against CD19-expressing K562 cells but similar in vivo anti-tumor efficacy with Lenti CAR-T. Taken together, these data provide insights into the phenotypic alterations induced by lentiviral transfection or transposon electroporation and will attract more attention to the clinical influence of different manufacturing procedures.

【 授权许可】

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Copyright © 2023 Niu, Zou, Hu, Zhang, Su, Wang, Lu, Zhang, Chen and Zhang