Modulation of intracellular kinase signaling to improve TIL stemness and function for adoptive cell therapy

Abstract Introduction Adoptive cellular therapy with tumor‐infiltrating lymphocytes (TIL) has demonstrated promising clinical benefits in several solid tumors, but the efficacy of this therapy might be compromised by the “prone‐to‐exhaustion” phenotype of TIL and poor persistence in vivo. This calls...

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Main Authors: Hao Feng, Ling Qiu, Zixiao Shi, Yao Sheng, Peipei Zhao, Di Zhou, Fei Li, Hailin Yu, Yanan You, Hui Wang, Ming Li, Shurong Zhu, Yan Du, Jun Cui, Jingwei Sun, Yarong Liu, Hua Jiang, Xin Wu
Format: Article
Language:English
Published: Wiley 2023-02-01
Series:Cancer Medicine
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Online Access:https://doi.org/10.1002/cam4.5095
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author Hao Feng
Ling Qiu
Zixiao Shi
Yao Sheng
Peipei Zhao
Di Zhou
Fei Li
Hailin Yu
Yanan You
Hui Wang
Ming Li
Shurong Zhu
Yan Du
Jun Cui
Jingwei Sun
Yarong Liu
Hua Jiang
Xin Wu
author_facet Hao Feng
Ling Qiu
Zixiao Shi
Yao Sheng
Peipei Zhao
Di Zhou
Fei Li
Hailin Yu
Yanan You
Hui Wang
Ming Li
Shurong Zhu
Yan Du
Jun Cui
Jingwei Sun
Yarong Liu
Hua Jiang
Xin Wu
author_sort Hao Feng
collection DOAJ
description Abstract Introduction Adoptive cellular therapy with tumor‐infiltrating lymphocytes (TIL) has demonstrated promising clinical benefits in several solid tumors, but the efficacy of this therapy might be compromised by the “prone‐to‐exhaustion” phenotype of TIL and poor persistence in vivo. This calls for a robust expansion process to produce a large number of cells for clinical usage while at the same time maintaining favorable anti‐tumor function and memory phenotype. Previous studies showed that the PI3K‐AKT signaling pathway plays a key role in the regulation of T cell activation, differentiation and memory formation. Method We modulated the PI3K‐AKT pathway in TIL isolated from cervical and ovarian cancer by application of AKT or PI3K inhibitors or CRISPR knockout of AKT1 and/or AKT2, and characterized their effects on TIL phenotype and effector function. Mechanistic study was further performed with RNA‐seq analysis of AKT1/2 KO TIL in comparison to control TIL. Result The inhibition of either PI3K or AKT led to an increase in the population of effector CD8+ T cells with upregulation of activation markers, elevated CD39−CD69− memory T cells, and significantly enhanced cytotoxicity when cocultured with tumor cell lines and patient‐derived tumor samples. Moreover, dual knockout of AKT1 and AKT2 largely phenocopies the functional impact of AKT or PI3K inhibition on TIL. This result was further validated by RNA‐seq analysis indicating that AKT1/2 ablation primarily regulates T cell differentiation and function‐related programs. Conclusion Modulation of PI3K‐AKT signaling represents a promising strategy to enhance TIL stemness and cytotoxicity and improve the clinical outcome of current TIL‐based therapy to treat solid tumors.
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spelling doaj-art-99fa77ab873e42639618fa69468134cb2025-08-20T01:53:22ZengWileyCancer Medicine2045-76342023-02-011233313332710.1002/cam4.5095Modulation of intracellular kinase signaling to improve TIL stemness and function for adoptive cell therapyHao Feng0Ling Qiu1Zixiao Shi2Yao Sheng3Peipei Zhao4Di Zhou5Fei Li6Hailin Yu7Yanan You8Hui Wang9Ming Li10Shurong Zhu11Yan Du12Jun Cui13Jingwei Sun14Yarong Liu15Hua Jiang16Xin Wu17Department of Gynecology Obstetrics and Gynecology Hospital of Fudan University Shanghai People's Republic of ChinaDepartment of Gynecology Obstetrics and Gynecology Hospital of Fudan University Shanghai People's Republic of ChinaGrit Biotechnology Co., Ltd. Shanghai Shanghai People's Republic of ChinaGrit Biotechnology Co., Ltd. Shanghai Shanghai People's Republic of ChinaGrit Biotechnology Co., Ltd. Shanghai Shanghai People's Republic of ChinaGrit Biotechnology Co., Ltd. Shanghai Shanghai People's Republic of ChinaGrit Biotechnology Co., Ltd. Shanghai Shanghai People's Republic of ChinaDepartment of Gynecology Obstetrics and Gynecology Hospital of Fudan University Shanghai People's Republic of ChinaDepartment of Gynecology Obstetrics and Gynecology Hospital of Fudan University Shanghai People's Republic of ChinaDepartment of Gynecology Obstetrics and Gynecology Hospital of Fudan University Shanghai People's Republic of ChinaDepartment of Gynecology Obstetrics and Gynecology Hospital of Fudan University Shanghai People's Republic of ChinaDepartment of Gynecology Obstetrics and Gynecology Hospital of Fudan University Shanghai People's Republic of ChinaDepartment of Gynecology Obstetrics and Gynecology Hospital of Fudan University Shanghai People's Republic of ChinaGrit Biotechnology Co., Ltd. Shanghai Shanghai People's Republic of ChinaGrit Biotechnology Co., Ltd. Shanghai Shanghai People's Republic of ChinaGrit Biotechnology Co., Ltd. Shanghai Shanghai People's Republic of ChinaDepartment of Gynecology Obstetrics and Gynecology Hospital of Fudan University Shanghai People's Republic of ChinaDepartment of Gynecology Obstetrics and Gynecology Hospital of Fudan University Shanghai People's Republic of ChinaAbstract Introduction Adoptive cellular therapy with tumor‐infiltrating lymphocytes (TIL) has demonstrated promising clinical benefits in several solid tumors, but the efficacy of this therapy might be compromised by the “prone‐to‐exhaustion” phenotype of TIL and poor persistence in vivo. This calls for a robust expansion process to produce a large number of cells for clinical usage while at the same time maintaining favorable anti‐tumor function and memory phenotype. Previous studies showed that the PI3K‐AKT signaling pathway plays a key role in the regulation of T cell activation, differentiation and memory formation. Method We modulated the PI3K‐AKT pathway in TIL isolated from cervical and ovarian cancer by application of AKT or PI3K inhibitors or CRISPR knockout of AKT1 and/or AKT2, and characterized their effects on TIL phenotype and effector function. Mechanistic study was further performed with RNA‐seq analysis of AKT1/2 KO TIL in comparison to control TIL. Result The inhibition of either PI3K or AKT led to an increase in the population of effector CD8+ T cells with upregulation of activation markers, elevated CD39−CD69− memory T cells, and significantly enhanced cytotoxicity when cocultured with tumor cell lines and patient‐derived tumor samples. Moreover, dual knockout of AKT1 and AKT2 largely phenocopies the functional impact of AKT or PI3K inhibition on TIL. This result was further validated by RNA‐seq analysis indicating that AKT1/2 ablation primarily regulates T cell differentiation and function‐related programs. Conclusion Modulation of PI3K‐AKT signaling represents a promising strategy to enhance TIL stemness and cytotoxicity and improve the clinical outcome of current TIL‐based therapy to treat solid tumors.https://doi.org/10.1002/cam4.5095AKT inhibitorAKT KOPI3K inhibitorTIL manufacturetumor‐infiltrating lymphocyte therapy
spellingShingle Hao Feng
Ling Qiu
Zixiao Shi
Yao Sheng
Peipei Zhao
Di Zhou
Fei Li
Hailin Yu
Yanan You
Hui Wang
Ming Li
Shurong Zhu
Yan Du
Jun Cui
Jingwei Sun
Yarong Liu
Hua Jiang
Xin Wu
Modulation of intracellular kinase signaling to improve TIL stemness and function for adoptive cell therapy
Cancer Medicine
AKT inhibitor
AKT KO
PI3K inhibitor
TIL manufacture
tumor‐infiltrating lymphocyte therapy
title Modulation of intracellular kinase signaling to improve TIL stemness and function for adoptive cell therapy
title_full Modulation of intracellular kinase signaling to improve TIL stemness and function for adoptive cell therapy
title_fullStr Modulation of intracellular kinase signaling to improve TIL stemness and function for adoptive cell therapy
title_full_unstemmed Modulation of intracellular kinase signaling to improve TIL stemness and function for adoptive cell therapy
title_short Modulation of intracellular kinase signaling to improve TIL stemness and function for adoptive cell therapy
title_sort modulation of intracellular kinase signaling to improve til stemness and function for adoptive cell therapy
topic AKT inhibitor
AKT KO
PI3K inhibitor
TIL manufacture
tumor‐infiltrating lymphocyte therapy
url https://doi.org/10.1002/cam4.5095
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