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

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 cur...

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Autores principales: Niu, Anna, Zou, Jintao, Hu, Xuan, Zhang, Zhang, Su, Lingyu, Wang, Jing, Lu, Xing, Zhang, Wei, Chen, Wei, Zhang, Xiaopeng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10203401/
https://www.ncbi.nlm.nih.gov/pubmed/37228617
http://dx.doi.org/10.3389/fimmu.2023.1068625
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author Niu, Anna
Zou, Jintao
Hu, Xuan
Zhang, Zhang
Su, Lingyu
Wang, Jing
Lu, Xing
Zhang, Wei
Chen, Wei
Zhang, Xiaopeng
author_facet Niu, Anna
Zou, Jintao
Hu, Xuan
Zhang, Zhang
Su, Lingyu
Wang, Jing
Lu, Xing
Zhang, Wei
Chen, Wei
Zhang, Xiaopeng
author_sort Niu, Anna
collection PubMed
description 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.
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spelling pubmed-102034012023-05-24 Differences in the phenotypes and transcriptomic signatures of chimeric antigen receptor T lymphocytes manufactured via electroporation or lentiviral transfection Niu, Anna Zou, Jintao Hu, Xuan Zhang, Zhang Su, Lingyu Wang, Jing Lu, Xing Zhang, Wei Chen, Wei Zhang, Xiaopeng Front Immunol Immunology 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. Frontiers Media S.A. 2023-05-09 /pmc/articles/PMC10203401/ /pubmed/37228617 http://dx.doi.org/10.3389/fimmu.2023.1068625 Text en Copyright © 2023 Niu, Zou, Hu, Zhang, Su, Wang, Lu, Zhang, Chen and Zhang https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Immunology
Niu, Anna
Zou, Jintao
Hu, Xuan
Zhang, Zhang
Su, Lingyu
Wang, Jing
Lu, Xing
Zhang, Wei
Chen, Wei
Zhang, Xiaopeng
Differences in the phenotypes and transcriptomic signatures of chimeric antigen receptor T lymphocytes manufactured via electroporation or lentiviral transfection
title Differences in the phenotypes and transcriptomic signatures of chimeric antigen receptor T lymphocytes manufactured via electroporation or lentiviral transfection
title_full Differences in the phenotypes and transcriptomic signatures of chimeric antigen receptor T lymphocytes manufactured via electroporation or lentiviral transfection
title_fullStr Differences in the phenotypes and transcriptomic signatures of chimeric antigen receptor T lymphocytes manufactured via electroporation or lentiviral transfection
title_full_unstemmed Differences in the phenotypes and transcriptomic signatures of chimeric antigen receptor T lymphocytes manufactured via electroporation or lentiviral transfection
title_short Differences in the phenotypes and transcriptomic signatures of chimeric antigen receptor T lymphocytes manufactured via electroporation or lentiviral transfection
title_sort differences in the phenotypes and transcriptomic signatures of chimeric antigen receptor t lymphocytes manufactured via electroporation or lentiviral transfection
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10203401/
https://www.ncbi.nlm.nih.gov/pubmed/37228617
http://dx.doi.org/10.3389/fimmu.2023.1068625
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