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TCR-independent Activation in Presence of a Src-family Kinase Inhibitor Improves CAR-T Cell Product Attributes

Chimeric antigen receptor expressing T cells (CAR-T cells) have shown remarkable efficacy against some blood cancers and have potential to treat many other human diseases. During CAR-T cell manufacturing, T cells are activated via engagement of the T-cell receptor (TCR); however, persistent TCR enga...

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Autores principales: Lamture, Gauri, Baer, Alan, Fischer, Joseph W., Colon-Moran, Winston, Bhattarai, Nirjal
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Lippincott Williams & Wilkins 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8906249/
https://www.ncbi.nlm.nih.gov/pubmed/34802014
http://dx.doi.org/10.1097/CJI.0000000000000402
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author Lamture, Gauri
Baer, Alan
Fischer, Joseph W.
Colon-Moran, Winston
Bhattarai, Nirjal
author_facet Lamture, Gauri
Baer, Alan
Fischer, Joseph W.
Colon-Moran, Winston
Bhattarai, Nirjal
author_sort Lamture, Gauri
collection PubMed
description Chimeric antigen receptor expressing T cells (CAR-T cells) have shown remarkable efficacy against some blood cancers and have potential to treat many other human diseases. During CAR-T cell manufacturing, T cells are activated via engagement of the T-cell receptor (TCR); however, persistent TCR engagement can induce unchecked activation, differentiation, and exhaustion, which can negatively affect CAR-T cell product quality and in vivo potency. In addition, T cells may not uniformly respond to TCR-dependent activation (TCR(D)) contributing to lot-to-lot variability, poor expansion, and manufacturing failures. TCR(D) also presents challenges during manufacturing of allogeneic CAR-T cells when endogenous TCR is deleted to prevent graft-versus-host disease. Thus, novel strategies to activate T cells may help improve CAR-T cell product attributes and reduce manufacturing failures. In this study, we compared the effect of TCR(D) and TCR-independent activation (TCR(I)) on CAR-T cell product attributes. We found that TCR(I) in presence of a Src-kinase inhibitor significantly improved CAR-T cell expansion and yield without affecting viability and CD4/CD8 ratio. Markers of T-cell activation, exhaustion and differentiation were also reduced in these CAR-T cells compared with CAR-T cells manufactured by TCR(D). TCR(I) did not affect CAR-T cell in vitro potency; however, following co-culture with target cells, CAR-T cells manufactured by TCR(I) released significantly less inflammatory cytokines compared with CAR-T cells manufactured by TCR(D). Together, these data suggest that manufacturing CAR-T cells by TCR(I) activation in the presence of a Src-kinase inhibitor improves product quality attributes and may help reduce manufacturing failures and improve CAR-T cell safety and efficacy in vivo.
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spelling pubmed-89062492022-03-10 TCR-independent Activation in Presence of a Src-family Kinase Inhibitor Improves CAR-T Cell Product Attributes Lamture, Gauri Baer, Alan Fischer, Joseph W. Colon-Moran, Winston Bhattarai, Nirjal J Immunother Basic Studies Chimeric antigen receptor expressing T cells (CAR-T cells) have shown remarkable efficacy against some blood cancers and have potential to treat many other human diseases. During CAR-T cell manufacturing, T cells are activated via engagement of the T-cell receptor (TCR); however, persistent TCR engagement can induce unchecked activation, differentiation, and exhaustion, which can negatively affect CAR-T cell product quality and in vivo potency. In addition, T cells may not uniformly respond to TCR-dependent activation (TCR(D)) contributing to lot-to-lot variability, poor expansion, and manufacturing failures. TCR(D) also presents challenges during manufacturing of allogeneic CAR-T cells when endogenous TCR is deleted to prevent graft-versus-host disease. Thus, novel strategies to activate T cells may help improve CAR-T cell product attributes and reduce manufacturing failures. In this study, we compared the effect of TCR(D) and TCR-independent activation (TCR(I)) on CAR-T cell product attributes. We found that TCR(I) in presence of a Src-kinase inhibitor significantly improved CAR-T cell expansion and yield without affecting viability and CD4/CD8 ratio. Markers of T-cell activation, exhaustion and differentiation were also reduced in these CAR-T cells compared with CAR-T cells manufactured by TCR(D). TCR(I) did not affect CAR-T cell in vitro potency; however, following co-culture with target cells, CAR-T cells manufactured by TCR(I) released significantly less inflammatory cytokines compared with CAR-T cells manufactured by TCR(D). Together, these data suggest that manufacturing CAR-T cells by TCR(I) activation in the presence of a Src-kinase inhibitor improves product quality attributes and may help reduce manufacturing failures and improve CAR-T cell safety and efficacy in vivo. Lippincott Williams & Wilkins 2022-04 2021-11-19 /pmc/articles/PMC8906249/ /pubmed/34802014 http://dx.doi.org/10.1097/CJI.0000000000000402 Text en Written work prepared by employees of the Federal Government as part of their official duties is, under the U.S. Copyright Act, a “work of the United States Government” for which copyright protection under Title 17 of the United States Code is not available. As such, copyright does not extend to the contributions of employees of the Federal Government.
spellingShingle Basic Studies
Lamture, Gauri
Baer, Alan
Fischer, Joseph W.
Colon-Moran, Winston
Bhattarai, Nirjal
TCR-independent Activation in Presence of a Src-family Kinase Inhibitor Improves CAR-T Cell Product Attributes
title TCR-independent Activation in Presence of a Src-family Kinase Inhibitor Improves CAR-T Cell Product Attributes
title_full TCR-independent Activation in Presence of a Src-family Kinase Inhibitor Improves CAR-T Cell Product Attributes
title_fullStr TCR-independent Activation in Presence of a Src-family Kinase Inhibitor Improves CAR-T Cell Product Attributes
title_full_unstemmed TCR-independent Activation in Presence of a Src-family Kinase Inhibitor Improves CAR-T Cell Product Attributes
title_short TCR-independent Activation in Presence of a Src-family Kinase Inhibitor Improves CAR-T Cell Product Attributes
title_sort tcr-independent activation in presence of a src-family kinase inhibitor improves car-t cell product attributes
topic Basic Studies
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8906249/
https://www.ncbi.nlm.nih.gov/pubmed/34802014
http://dx.doi.org/10.1097/CJI.0000000000000402
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