CD28 is superior to 4-1BB costimulation in generating CAR-NK cells for tumor immunotherapy

Abstract Chimeric antigen receptor (CAR)-NK therapy holds great potential for tumor treatment, but current CAR designs are primarily optimized for T cells, raising concerns about their suitability for NK cells. This study compared two dominant CAR designs used in T cells—CD28-CD3ζ (28z) and 4-1BB-CD...

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Main Authors: Pengchao Zhang, Xuejia Feng, Xiangyun Niu, Zhongming Liu, Minghui Li, Maoxuan Liu, Dehong Yan, Guizhong Zhang, Xiaochun Wan
Format: Article
Language:English
Published: BMC 2025-03-01
Series:Experimental Hematology & Oncology
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Online Access:https://doi.org/10.1186/s40164-025-00618-7
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author Pengchao Zhang
Xuejia Feng
Xiangyun Niu
Zhongming Liu
Minghui Li
Maoxuan Liu
Dehong Yan
Guizhong Zhang
Xiaochun Wan
author_facet Pengchao Zhang
Xuejia Feng
Xiangyun Niu
Zhongming Liu
Minghui Li
Maoxuan Liu
Dehong Yan
Guizhong Zhang
Xiaochun Wan
author_sort Pengchao Zhang
collection DOAJ
description Abstract Chimeric antigen receptor (CAR)-NK therapy holds great potential for tumor treatment, but current CAR designs are primarily optimized for T cells, raising concerns about their suitability for NK cells. This study compared two dominant CAR designs used in T cells—CD28-CD3ζ (28z) and 4-1BB-CD3ζ (BBz)—and found that CD28 costimulation offers superior functionality in NK cells. 28z CAR-NK cells exhibited significantly better activation, cytotoxicity, and in vivo anti-tumor efficacy than BBz CAR-NK cells, with similar persistence and tumor infiltration. 28z CAR more effectively recruited the ZAP70 kinase and upregulated multiple key factors involved in immune activation, potentially augmenting CAR-NK cell function. MAP3K8, a kinase involved in inflammation and the MAPK signaling pathway, was identified as a critical mediator in enhancing 28z CAR-NK cell function. Silencing or inhibiting MAP3K8 impaired the anti-tumor activity of 28z CAR-NK cells, while its overexpression substantially improved the function of BBz CAR-NK cells. These findings provide new insights into how CD28 costimulation boosts CAR-NK cell efficacy, supporting its use into NK cell-specific CARs for cancer immunotherapy, and highlight MAP3K8 as a potential target for optimizing BBz CAR-NK cell therapy.
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issn 2162-3619
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publishDate 2025-03-01
publisher BMC
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series Experimental Hematology & Oncology
spelling doaj-art-d6ee4d307e544088b58c0e9a9cec8e822025-08-20T02:59:23ZengBMCExperimental Hematology & Oncology2162-36192025-03-011411510.1186/s40164-025-00618-7CD28 is superior to 4-1BB costimulation in generating CAR-NK cells for tumor immunotherapyPengchao Zhang0Xuejia Feng1Xiangyun Niu2Zhongming Liu3Minghui Li4Maoxuan Liu5Dehong Yan6Guizhong Zhang7Xiaochun Wan8Center for Protein and Cell-based Drugs, Institute of Biomedicine and Biotechnology, Shenzhen Institutes of Advanced Technology, Chinese Academy of SciencesCenter for Protein and Cell-based Drugs, Institute of Biomedicine and Biotechnology, Shenzhen Institutes of Advanced Technology, Chinese Academy of SciencesCenter for Protein and Cell-based Drugs, Institute of Biomedicine and Biotechnology, Shenzhen Institutes of Advanced Technology, Chinese Academy of SciencesCenter for Protein and Cell-based Drugs, Institute of Biomedicine and Biotechnology, Shenzhen Institutes of Advanced Technology, Chinese Academy of SciencesCenter for Protein and Cell-based Drugs, Institute of Biomedicine and Biotechnology, Shenzhen Institutes of Advanced Technology, Chinese Academy of SciencesCenter for Protein and Cell-based Drugs, Institute of Biomedicine and Biotechnology, Shenzhen Institutes of Advanced Technology, Chinese Academy of SciencesCenter for Protein and Cell-based Drugs, Institute of Biomedicine and Biotechnology, Shenzhen Institutes of Advanced Technology, Chinese Academy of SciencesCenter for Protein and Cell-based Drugs, Institute of Biomedicine and Biotechnology, Shenzhen Institutes of Advanced Technology, Chinese Academy of SciencesCenter for Protein and Cell-based Drugs, Institute of Biomedicine and Biotechnology, Shenzhen Institutes of Advanced Technology, Chinese Academy of SciencesAbstract Chimeric antigen receptor (CAR)-NK therapy holds great potential for tumor treatment, but current CAR designs are primarily optimized for T cells, raising concerns about their suitability for NK cells. This study compared two dominant CAR designs used in T cells—CD28-CD3ζ (28z) and 4-1BB-CD3ζ (BBz)—and found that CD28 costimulation offers superior functionality in NK cells. 28z CAR-NK cells exhibited significantly better activation, cytotoxicity, and in vivo anti-tumor efficacy than BBz CAR-NK cells, with similar persistence and tumor infiltration. 28z CAR more effectively recruited the ZAP70 kinase and upregulated multiple key factors involved in immune activation, potentially augmenting CAR-NK cell function. MAP3K8, a kinase involved in inflammation and the MAPK signaling pathway, was identified as a critical mediator in enhancing 28z CAR-NK cell function. Silencing or inhibiting MAP3K8 impaired the anti-tumor activity of 28z CAR-NK cells, while its overexpression substantially improved the function of BBz CAR-NK cells. These findings provide new insights into how CD28 costimulation boosts CAR-NK cell efficacy, supporting its use into NK cell-specific CARs for cancer immunotherapy, and highlight MAP3K8 as a potential target for optimizing BBz CAR-NK cell therapy.https://doi.org/10.1186/s40164-025-00618-7CAR-NKTranscriptomicsCD284-1BBMAP3K8
spellingShingle Pengchao Zhang
Xuejia Feng
Xiangyun Niu
Zhongming Liu
Minghui Li
Maoxuan Liu
Dehong Yan
Guizhong Zhang
Xiaochun Wan
CD28 is superior to 4-1BB costimulation in generating CAR-NK cells for tumor immunotherapy
Experimental Hematology & Oncology
CAR-NK
Transcriptomics
CD28
4-1BB
MAP3K8
title CD28 is superior to 4-1BB costimulation in generating CAR-NK cells for tumor immunotherapy
title_full CD28 is superior to 4-1BB costimulation in generating CAR-NK cells for tumor immunotherapy
title_fullStr CD28 is superior to 4-1BB costimulation in generating CAR-NK cells for tumor immunotherapy
title_full_unstemmed CD28 is superior to 4-1BB costimulation in generating CAR-NK cells for tumor immunotherapy
title_short CD28 is superior to 4-1BB costimulation in generating CAR-NK cells for tumor immunotherapy
title_sort cd28 is superior to 4 1bb costimulation in generating car nk cells for tumor immunotherapy
topic CAR-NK
Transcriptomics
CD28
4-1BB
MAP3K8
url https://doi.org/10.1186/s40164-025-00618-7
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