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Structure of the Signal Transduction Domain in Second-Generation CAR Regulates the Input Efficiency of CAR Signals

T cells that are genetically engineered to express chimeric antigen receptor (CAR) have a strong potential to eliminate tumor cells, yet the CAR-T cells may also induce severe side effects due to an excessive immune response. Although optimization of the CAR structure is expected to improve the effi...

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Autores principales: Fujiwara, Kento, Kitaura, Masaki, Tsunei, Ayaka, Kusabuka, Hotaka, Ogaki, Erika, Okada, Naoki
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7957710/
https://www.ncbi.nlm.nih.gov/pubmed/33804441
http://dx.doi.org/10.3390/ijms22052476
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author Fujiwara, Kento
Kitaura, Masaki
Tsunei, Ayaka
Kusabuka, Hotaka
Ogaki, Erika
Okada, Naoki
author_facet Fujiwara, Kento
Kitaura, Masaki
Tsunei, Ayaka
Kusabuka, Hotaka
Ogaki, Erika
Okada, Naoki
author_sort Fujiwara, Kento
collection PubMed
description T cells that are genetically engineered to express chimeric antigen receptor (CAR) have a strong potential to eliminate tumor cells, yet the CAR-T cells may also induce severe side effects due to an excessive immune response. Although optimization of the CAR structure is expected to improve the efficacy and toxicity of CAR-T cells, the relationship between CAR structure and CAR-T cell functions remains unclear. Here, we constructed second-generation CARs incorporating a signal transduction domain (STD) derived from CD3ζ and a 2nd STD derived from CD28, CD278, CD27, CD134, or CD137, and investigated the impact of the STD structure and signaling on CAR-T cell functions. Cytokine secretion of CAR-T cells was enhanced by 2nd STD signaling. T cells expressing CAR with CD278-STD or CD137-STD proliferated in an antigen-independent manner by their STD tonic signaling. CAR-T cells incorporating CD28-STD or CD278-STD between TMD and CD3ζ-STD showed higher cytotoxicity than first-generation CAR or second-generation CARs with other 2nd STDs. The potent cytotoxicity of these CAR-T cells was not affected by inhibiting the 2nd STD signals, but was eliminated by placing the STDs after the CD3ζ-STD. Our data highlighted that CAR activity was affected by STD structure as well as by 2nd STD signaling.
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spelling pubmed-79577102021-03-16 Structure of the Signal Transduction Domain in Second-Generation CAR Regulates the Input Efficiency of CAR Signals Fujiwara, Kento Kitaura, Masaki Tsunei, Ayaka Kusabuka, Hotaka Ogaki, Erika Okada, Naoki Int J Mol Sci Article T cells that are genetically engineered to express chimeric antigen receptor (CAR) have a strong potential to eliminate tumor cells, yet the CAR-T cells may also induce severe side effects due to an excessive immune response. Although optimization of the CAR structure is expected to improve the efficacy and toxicity of CAR-T cells, the relationship between CAR structure and CAR-T cell functions remains unclear. Here, we constructed second-generation CARs incorporating a signal transduction domain (STD) derived from CD3ζ and a 2nd STD derived from CD28, CD278, CD27, CD134, or CD137, and investigated the impact of the STD structure and signaling on CAR-T cell functions. Cytokine secretion of CAR-T cells was enhanced by 2nd STD signaling. T cells expressing CAR with CD278-STD or CD137-STD proliferated in an antigen-independent manner by their STD tonic signaling. CAR-T cells incorporating CD28-STD or CD278-STD between TMD and CD3ζ-STD showed higher cytotoxicity than first-generation CAR or second-generation CARs with other 2nd STDs. The potent cytotoxicity of these CAR-T cells was not affected by inhibiting the 2nd STD signals, but was eliminated by placing the STDs after the CD3ζ-STD. Our data highlighted that CAR activity was affected by STD structure as well as by 2nd STD signaling. MDPI 2021-03-01 /pmc/articles/PMC7957710/ /pubmed/33804441 http://dx.doi.org/10.3390/ijms22052476 Text en © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Fujiwara, Kento
Kitaura, Masaki
Tsunei, Ayaka
Kusabuka, Hotaka
Ogaki, Erika
Okada, Naoki
Structure of the Signal Transduction Domain in Second-Generation CAR Regulates the Input Efficiency of CAR Signals
title Structure of the Signal Transduction Domain in Second-Generation CAR Regulates the Input Efficiency of CAR Signals
title_full Structure of the Signal Transduction Domain in Second-Generation CAR Regulates the Input Efficiency of CAR Signals
title_fullStr Structure of the Signal Transduction Domain in Second-Generation CAR Regulates the Input Efficiency of CAR Signals
title_full_unstemmed Structure of the Signal Transduction Domain in Second-Generation CAR Regulates the Input Efficiency of CAR Signals
title_short Structure of the Signal Transduction Domain in Second-Generation CAR Regulates the Input Efficiency of CAR Signals
title_sort structure of the signal transduction domain in second-generation car regulates the input efficiency of car signals
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7957710/
https://www.ncbi.nlm.nih.gov/pubmed/33804441
http://dx.doi.org/10.3390/ijms22052476
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