Cord blood-derived iNK T cells as a platform for allogeneic CAR T cell therapy

CD1d-restricted invariant Natural Killer (iNK) T cells are a suitable candidate for allogeneic Chimeric Antigen Receptor (CAR) T cell therapy as they do not cause graft-versus-host disease (GvHD) due to the monomorphic nature of CD1d proteins. However, the phenotypic and functional heterogeneity of...

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Main Authors: Maison Grefe, Abel Trujillo-Ocampo, Jelita Clinton, Hong He, Ling Yu, Dan Li, Qing Ma, Elizabeth J. Shpall, Jeffrey J. Molldrem, Jin S. Im
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-05-01
Series:Frontiers in Immunology
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Online Access:https://www.frontiersin.org/articles/10.3389/fimmu.2025.1621260/full
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author Maison Grefe
Abel Trujillo-Ocampo
Jelita Clinton
Hong He
Ling Yu
Dan Li
Qing Ma
Elizabeth J. Shpall
Jeffrey J. Molldrem
Jin S. Im
Jin S. Im
author_facet Maison Grefe
Abel Trujillo-Ocampo
Jelita Clinton
Hong He
Ling Yu
Dan Li
Qing Ma
Elizabeth J. Shpall
Jeffrey J. Molldrem
Jin S. Im
Jin S. Im
author_sort Maison Grefe
collection DOAJ
description CD1d-restricted invariant Natural Killer (iNK) T cells are a suitable candidate for allogeneic Chimeric Antigen Receptor (CAR) T cell therapy as they do not cause graft-versus-host disease (GvHD) due to the monomorphic nature of CD1d proteins. However, the phenotypic and functional heterogeneity of iNK T cells from adult donors (AD) may lead to the inconstant CAR-iNK T cell products. Cord blood-derived (CB) iNK T cells, in contrast, exhibit inter-donor homogeneity in phenotype including uniform CD4 expression and are enriched in memory iNK T cell populations. Thus, we evaluated the preclinical therapeutic potential of iNK T cells derived from cord blood (CB) as an off the shelf CAR T cell therapy platform, given the dominant presence of CD4+ iNK T cells. First, CB-derived iNK T cells were extremely enriched with CD4+ iNK T cells that express various NK receptors and display iNK-TCR mediated cytotoxicity but in a lesser degree than AD-derived CD4- iNK T cells. When engineered with an 8F4CAR targeting the acute myeloid leukemia-associated antigen PR1 presented in HLA-A2*01, CB-8F4CAR-iNK T cells showed a greater expansion capacity with higher CD62L expression than AD-8F4CAR-iNK T cells but with similar 8F4CAR expression and iNK T purity. CB-8F4CAR-iNK T cells displayed in vitro cytotoxicity against PR1/HLA-A2+ primary Acute Myeloid Leukemia (AML) and cell lines better than AD-8F4CAR iNK T cells and maintained potent cytotoxicity in repeated antigenic challenges. Moreover, CB-8F4CAR-iNK T cells showed anti-leukemia activity in vivo in a dose dependent manner. Lastly, CB-8F4CAR-iNK T cells were polarized to produce Th2-biased cytokines but in a lesser amount after 8F4CAR-mediated leukemia cytolysis compared to iNK-TCR mediated activation. In conclusion, consistent CD4+ phenotype, superior expansion capacity, and enhanced CD62L expression of CB-CAR-iNK T cells suggest that they may provide an alternative off-the-shelf source for effective CAR-iNK T cell therapy, while reducing the risk of severe cytokine release syndrome through their immunomodulatory properties. Thus, our results support the potential use of CB-iNK T cells as an allogeneic CAR-T cell therapy platform as they maintain a potent cytotoxicity with potentially better safety profile given a Th2-biased cytokine production upon activation.
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spelling doaj-art-0db22e8251bd4706b496a2676d97176a2025-08-20T02:16:49ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-05-011610.3389/fimmu.2025.16212601621260Cord blood-derived iNK T cells as a platform for allogeneic CAR T cell therapyMaison Grefe0Abel Trujillo-Ocampo1Jelita Clinton2Hong He3Ling Yu4Dan Li5Qing Ma6Elizabeth J. Shpall7Jeffrey J. Molldrem8Jin S. Im9Jin S. Im10Department of Hematopoietic Biology & Malignancies, The University of Texas MD Anderson Cancer Center, Houston, TX, United StatesDepartment of Hematopoietic Biology & Malignancies, The University of Texas MD Anderson Cancer Center, Houston, TX, United StatesDepartment of Hematopoietic Biology & Malignancies, The University of Texas MD Anderson Cancer Center, Houston, TX, United StatesDepartment of Hematopoietic Biology & Malignancies, The University of Texas MD Anderson Cancer Center, Houston, TX, United StatesDepartment of Hematopoietic Biology & Malignancies, The University of Texas MD Anderson Cancer Center, Houston, TX, United StatesDepartment of Hematopoietic Biology & Malignancies, The University of Texas MD Anderson Cancer Center, Houston, TX, United StatesDepartment of Hematopoietic Biology & Malignancies, The University of Texas MD Anderson Cancer Center, Houston, TX, United StatesDepartment of Stem Cell Transplantation & Cellular Therapy, The University of Texas MD Anderson Cancer Center, Houston, TX, United StatesDepartment of Hematopoietic Biology & Malignancies, The University of Texas MD Anderson Cancer Center, Houston, TX, United StatesDepartment of Hematopoietic Biology & Malignancies, The University of Texas MD Anderson Cancer Center, Houston, TX, United StatesDepartment of Stem Cell Transplantation & Cellular Therapy, The University of Texas MD Anderson Cancer Center, Houston, TX, United StatesCD1d-restricted invariant Natural Killer (iNK) T cells are a suitable candidate for allogeneic Chimeric Antigen Receptor (CAR) T cell therapy as they do not cause graft-versus-host disease (GvHD) due to the monomorphic nature of CD1d proteins. However, the phenotypic and functional heterogeneity of iNK T cells from adult donors (AD) may lead to the inconstant CAR-iNK T cell products. Cord blood-derived (CB) iNK T cells, in contrast, exhibit inter-donor homogeneity in phenotype including uniform CD4 expression and are enriched in memory iNK T cell populations. Thus, we evaluated the preclinical therapeutic potential of iNK T cells derived from cord blood (CB) as an off the shelf CAR T cell therapy platform, given the dominant presence of CD4+ iNK T cells. First, CB-derived iNK T cells were extremely enriched with CD4+ iNK T cells that express various NK receptors and display iNK-TCR mediated cytotoxicity but in a lesser degree than AD-derived CD4- iNK T cells. When engineered with an 8F4CAR targeting the acute myeloid leukemia-associated antigen PR1 presented in HLA-A2*01, CB-8F4CAR-iNK T cells showed a greater expansion capacity with higher CD62L expression than AD-8F4CAR-iNK T cells but with similar 8F4CAR expression and iNK T purity. CB-8F4CAR-iNK T cells displayed in vitro cytotoxicity against PR1/HLA-A2+ primary Acute Myeloid Leukemia (AML) and cell lines better than AD-8F4CAR iNK T cells and maintained potent cytotoxicity in repeated antigenic challenges. Moreover, CB-8F4CAR-iNK T cells showed anti-leukemia activity in vivo in a dose dependent manner. Lastly, CB-8F4CAR-iNK T cells were polarized to produce Th2-biased cytokines but in a lesser amount after 8F4CAR-mediated leukemia cytolysis compared to iNK-TCR mediated activation. In conclusion, consistent CD4+ phenotype, superior expansion capacity, and enhanced CD62L expression of CB-CAR-iNK T cells suggest that they may provide an alternative off-the-shelf source for effective CAR-iNK T cell therapy, while reducing the risk of severe cytokine release syndrome through their immunomodulatory properties. Thus, our results support the potential use of CB-iNK T cells as an allogeneic CAR-T cell therapy platform as they maintain a potent cytotoxicity with potentially better safety profile given a Th2-biased cytokine production upon activation.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1621260/fullinvariant natural killer t cellscord bloodchimeric antigen receptoracute myeloid leukemiaCAR T cell therapy
spellingShingle Maison Grefe
Abel Trujillo-Ocampo
Jelita Clinton
Hong He
Ling Yu
Dan Li
Qing Ma
Elizabeth J. Shpall
Jeffrey J. Molldrem
Jin S. Im
Jin S. Im
Cord blood-derived iNK T cells as a platform for allogeneic CAR T cell therapy
Frontiers in Immunology
invariant natural killer t cells
cord blood
chimeric antigen receptor
acute myeloid leukemia
CAR T cell therapy
title Cord blood-derived iNK T cells as a platform for allogeneic CAR T cell therapy
title_full Cord blood-derived iNK T cells as a platform for allogeneic CAR T cell therapy
title_fullStr Cord blood-derived iNK T cells as a platform for allogeneic CAR T cell therapy
title_full_unstemmed Cord blood-derived iNK T cells as a platform for allogeneic CAR T cell therapy
title_short Cord blood-derived iNK T cells as a platform for allogeneic CAR T cell therapy
title_sort cord blood derived ink t cells as a platform for allogeneic car t cell therapy
topic invariant natural killer t cells
cord blood
chimeric antigen receptor
acute myeloid leukemia
CAR T cell therapy
url https://www.frontiersin.org/articles/10.3389/fimmu.2025.1621260/full
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