Co-expression of IL-15 and CCL21 strengthens CAR-NK cells to eliminate tumors in concert with T cells and equips them with PI3K/AKT/mTOR signal signature

Background Chimeric antigen receptor (CAR)-natural killer (NK) cell therapy has demonstrated safety and feasibility in clinical settings; however, limited efficacy due to intrinsic dysfunction and extrinsic suppression remains an unresolved issue. T cells provide multifaceted support to NK cell-medi...

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Main Authors: Jie Zhou, Lu Tang, Yu Hu, Jun Deng, Yingying Li, Wei Xiong, Jianghua Wu, Jia Xu, Wenjing Luo, Heng Mei, Xindi Wang, Zhaozhao Chen, Chenggong Li, Zhongpei Huang, Zhuolin Wu, Yinqiang Zhang, Yun Kang, Qiaolin Liu
Format: Article
Language:English
Published: BMJ Publishing Group 2025-06-01
Series:Journal for ImmunoTherapy of Cancer
Online Access:https://jitc.bmj.com/content/13/6/e010822.full
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author Jie Zhou
Lu Tang
Yu Hu
Jun Deng
Yingying Li
Wei Xiong
Jianghua Wu
Jia Xu
Wenjing Luo
Heng Mei
Xindi Wang
Zhaozhao Chen
Chenggong Li
Zhongpei Huang
Zhuolin Wu
Yinqiang Zhang
Yun Kang
Qiaolin Liu
author_facet Jie Zhou
Lu Tang
Yu Hu
Jun Deng
Yingying Li
Wei Xiong
Jianghua Wu
Jia Xu
Wenjing Luo
Heng Mei
Xindi Wang
Zhaozhao Chen
Chenggong Li
Zhongpei Huang
Zhuolin Wu
Yinqiang Zhang
Yun Kang
Qiaolin Liu
author_sort Jie Zhou
collection DOAJ
description Background Chimeric antigen receptor (CAR)-natural killer (NK) cell therapy has demonstrated safety and feasibility in clinical settings; however, limited efficacy due to intrinsic dysfunction and extrinsic suppression remains an unresolved issue. T cells provide multifaceted support to NK cell-mediated responses. Here, we aimed to design a novel CD19-targeted CAR-NK, engineered with secreted interleukin-15 and C-C motif chemokine ligand 21 (ie, 15×21 CAR-NK), capable of recruiting and cooperating with T cells.Methods We characterized 15×21 CAR-NK cells by performing experiments in vitro and in mouse models, and conducting RNA sequencing.Results 15×21 CAR-NK cells exhibit strong capabilities in cytotoxicity, cytokine production, effector molecule expression, and T-cell recruitment in vitro. Cooperation with T cells promoted efficient tumor-cell elimination, alleviated mutual exhaustion phenotypes, and enhanced the expression of effector molecules/receptors. The recruitment and cooperative effects also result in effective tumor control in mouse models. In addition, 15×21 CAR-NK cells strongly enrich the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway - a key intracellular signaling cascade that is associated with enhanced downstream pro-survival signaling, anti-apoptotic ability, mitochondrial function, and cytotoxicity.Conclusions Our study highlights the intrinsic advantages and extrinsic T-cell cooperative benefits of 15×21 CAR-NK cells, providing a promising strategy for NK-cell-based immunotherapy.
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spelling doaj-art-c9cfc013e4544667828d87bbb03869d12025-08-20T02:10:21ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262025-06-0113610.1136/jitc-2024-010822Co-expression of IL-15 and CCL21 strengthens CAR-NK cells to eliminate tumors in concert with T cells and equips them with PI3K/AKT/mTOR signal signatureJie Zhou0Lu Tang1Yu Hu2Jun Deng3Yingying Li4Wei Xiong5Jianghua Wu6Jia Xu7Wenjing Luo8Heng Mei9Xindi Wang10Zhaozhao Chen11Chenggong Li12Zhongpei Huang13Zhuolin Wu14Yinqiang Zhang15Yun Kang16Qiaolin Liu171 Department of Gastroenterology, Renmin Hospital of Wuhan University, Wuhan, ChinaDepartment of Stomatology, PLA 960th Hospital, Jinan, Shandong, ChinaKey Laboratory of Biological Targeted Therapy (Huazhong University of Science and Technology), Ministry of Education, Wuhan, Hubei, ChinaHubei Clinical Medical Center of Cell Therapy for Neoplastic Disease, Wuhan, Hubei, ChinaInstitute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, ChinaHubei Clinical Medical Center of Cell Therapy for Neoplastic Disease, Wuhan, Hubei, ChinaDepartment of Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong, ChinaDepartment of Radiology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, People`s Republic of China29 Department of Neurology, Wuhan General Hospital of PLA, Wuhan, ChinaKey Laboratory of Biological Targeted Therapy (Huazhong University of Science and Technology), Ministry of Education, Wuhan, Hubei, ChinaInstitute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, ChinaInstitute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, ChinaInstitute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, ChinaHubei Clinical Medical Center of Cell Therapy for Neoplastic Disease, Wuhan, Hubei, ChinaInstitute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, ChinaInstitute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, ChinaInstitute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, ChinaInstitute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, ChinaBackground Chimeric antigen receptor (CAR)-natural killer (NK) cell therapy has demonstrated safety and feasibility in clinical settings; however, limited efficacy due to intrinsic dysfunction and extrinsic suppression remains an unresolved issue. T cells provide multifaceted support to NK cell-mediated responses. Here, we aimed to design a novel CD19-targeted CAR-NK, engineered with secreted interleukin-15 and C-C motif chemokine ligand 21 (ie, 15×21 CAR-NK), capable of recruiting and cooperating with T cells.Methods We characterized 15×21 CAR-NK cells by performing experiments in vitro and in mouse models, and conducting RNA sequencing.Results 15×21 CAR-NK cells exhibit strong capabilities in cytotoxicity, cytokine production, effector molecule expression, and T-cell recruitment in vitro. Cooperation with T cells promoted efficient tumor-cell elimination, alleviated mutual exhaustion phenotypes, and enhanced the expression of effector molecules/receptors. The recruitment and cooperative effects also result in effective tumor control in mouse models. In addition, 15×21 CAR-NK cells strongly enrich the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway - a key intracellular signaling cascade that is associated with enhanced downstream pro-survival signaling, anti-apoptotic ability, mitochondrial function, and cytotoxicity.Conclusions Our study highlights the intrinsic advantages and extrinsic T-cell cooperative benefits of 15×21 CAR-NK cells, providing a promising strategy for NK-cell-based immunotherapy.https://jitc.bmj.com/content/13/6/e010822.full
spellingShingle Jie Zhou
Lu Tang
Yu Hu
Jun Deng
Yingying Li
Wei Xiong
Jianghua Wu
Jia Xu
Wenjing Luo
Heng Mei
Xindi Wang
Zhaozhao Chen
Chenggong Li
Zhongpei Huang
Zhuolin Wu
Yinqiang Zhang
Yun Kang
Qiaolin Liu
Co-expression of IL-15 and CCL21 strengthens CAR-NK cells to eliminate tumors in concert with T cells and equips them with PI3K/AKT/mTOR signal signature
Journal for ImmunoTherapy of Cancer
title Co-expression of IL-15 and CCL21 strengthens CAR-NK cells to eliminate tumors in concert with T cells and equips them with PI3K/AKT/mTOR signal signature
title_full Co-expression of IL-15 and CCL21 strengthens CAR-NK cells to eliminate tumors in concert with T cells and equips them with PI3K/AKT/mTOR signal signature
title_fullStr Co-expression of IL-15 and CCL21 strengthens CAR-NK cells to eliminate tumors in concert with T cells and equips them with PI3K/AKT/mTOR signal signature
title_full_unstemmed Co-expression of IL-15 and CCL21 strengthens CAR-NK cells to eliminate tumors in concert with T cells and equips them with PI3K/AKT/mTOR signal signature
title_short Co-expression of IL-15 and CCL21 strengthens CAR-NK cells to eliminate tumors in concert with T cells and equips them with PI3K/AKT/mTOR signal signature
title_sort co expression of il 15 and ccl21 strengthens car nk cells to eliminate tumors in concert with t cells and equips them with pi3k akt mtor signal signature
url https://jitc.bmj.com/content/13/6/e010822.full
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