eEF2K promotes PD-L1 stabilization through inactivating GSK3β in melanoma

Background Immune checkpoint blockade (ICB) targeting programmed death ligand-1 (PD-L1)/programmed cell death protein-1 (PD-1) pathway has become an attractive strategy for cancer treatment; however, unsatisfactory efficacy has limited its clinical benefits. Therefore, a more comprehensive understan...

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Main Authors: Jianxun Song, Xiang Chen, Ying Wen, Mingzhu Yin, Yan Cheng, Wenjun Yi, Xingcong Ren, Xisha Chen, Kuansong Wang, Shilong Jiang, Hongyin Sun, Xuanling Che, Minghui Zhang, Jiaying He, Mengting Liao, Xiangling Li, Xiaoming Zhou, Jinming Yang
Format: Article
Language:English
Published: BMJ Publishing Group 2022-03-01
Series:Journal for ImmunoTherapy of Cancer
Online Access:https://jitc.bmj.com/content/10/3/e004026.full
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author Jianxun Song
Xiang Chen
Ying Wen
Mingzhu Yin
Yan Cheng
Wenjun Yi
Xingcong Ren
Xisha Chen
Kuansong Wang
Shilong Jiang
Hongyin Sun
Xuanling Che
Minghui Zhang
Jiaying He
Mengting Liao
Xiangling Li
Xiaoming Zhou
Jinming Yang
author_facet Jianxun Song
Xiang Chen
Ying Wen
Mingzhu Yin
Yan Cheng
Wenjun Yi
Xingcong Ren
Xisha Chen
Kuansong Wang
Shilong Jiang
Hongyin Sun
Xuanling Che
Minghui Zhang
Jiaying He
Mengting Liao
Xiangling Li
Xiaoming Zhou
Jinming Yang
author_sort Jianxun Song
collection DOAJ
description Background Immune checkpoint blockade (ICB) targeting programmed death ligand-1 (PD-L1)/programmed cell death protein-1 (PD-1) pathway has become an attractive strategy for cancer treatment; however, unsatisfactory efficacy has limited its clinical benefits. Therefore, a more comprehensive understanding of the regulation of PD-L1 expression is essential for developing more effective cancer immunotherapy. Recent studies have revealed the important roles of eukaryotic elongation factor 2 kinase (eEF2K) in promoting epithelial-mesenchymal transition (EMT), angiogenesis, tumor cell migration and invasion; nevertheless, the exact role of eEF2K in the regulation of tumor immune microenvironment (TIME) remains largely unknown.Methods In this study, we used a cohort of 38 patients with melanoma who received anti-PD-1 treatment to explore the association between eEF2K expression and immunotherapy efficacy against melanoma. Immunoprecipitation-mass spectrometry analysis and in vitro assays were used to examine the role and molecular mechanism of eEF2K in regulating PD-L1 expression. We also determined the effects of eEF2K on tumor growth and cytotoxicity of CD8+ T cells in TIME in a mouse melanoma model. We further investigated the efficacy of the eEF2K inhibition in combination with anti-PD-1 treatment in vivo.Results High eEF2K expression is correlated with better therapeutic response and longer survival in patients with melanoma treated with PD-1 monoclonal antibody (mAb). Moreover, eEF2K protein expression is positively correlated with PD-L1 protein expression. Mechanistically, eEF2K directly bound to and inactivated glycogen synthase kinase 3 beta (GSK3β) by phosphorylating it at serine 9 (S9), leading to PD-L1 protein stabilization and upregulation, and subsequently tumor immune evasion. Knockdown of eEF2K decreased PD-L1 expression and enhanced CD8+ T cell activity, thus dramatically attenuating murine B16F10 melanoma growth in vivo. Clinically, p-GSK3β/S9 expression is positively correlated with the expressions of eEF2K and PD-L1, and the response to anti-PD-1 immunotherapy. Furthermore, eEF2K inhibitor, NH125 treatment or eEF2K knockdown enhanced the efficacy of PD-1 mAb therapy in a melanoma mouse model.Conclusions Our results suggest that eEF2K may serve as a biomarker for predicting therapeutic response and prognosis in patients receiving anti-PD-1 therapy, reveal a vital role of eEF2K in regulating TIME by controlling PD-L1 expression and provide a potential combination therapeutic strategy of eEF2K inhibition with ICB therapy.
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series Journal for ImmunoTherapy of Cancer
spelling doaj-art-0bcedc5a9af045bf95383a50330c16732025-08-20T03:05:14ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262022-03-0110310.1136/jitc-2021-004026eEF2K promotes PD-L1 stabilization through inactivating GSK3β in melanomaJianxun Song0Xiang Chen1Ying Wen2Mingzhu Yin3Yan Cheng4Wenjun Yi5Xingcong Ren6Xisha Chen7Kuansong Wang8Shilong Jiang9Hongyin Sun10Xuanling Che11Minghui Zhang12Jiaying He13Mengting Liao14Xiangling Li15Xiaoming Zhou16Jinming Yang17Microbial Pathogenesis and Immunology, Texas A&M University Health Sciences Center, Bryan, Texas, USADepartment of Dermatology, Xiangya Hospital, Central South University, Changsha, Hunan, ChinaDepartment of Communicable Disease Control and Prevention, Shenzhen Center for Disease Control and Prevention, Shenzhen, Guangdong, ChinaDepartment of Dermatology, Hunan Engineering Research Center of Skin Health and Disease, Hunan Key Laboratory of Skin Cancer and Psoriasis, Xiangya Hospital, Central South University, Changsha, ChinaHunan Provincial Engineering Research Centre of Translational Medicine and Innovative Drug, Changsha, ChinaDepartment of General Surgery, The Second Xiangya Hospital, Central South University, Changsha, ChinaToxicology and Cancer Biology, University of Kentucky, Lexington, Kentucky, USAHunan Provincial Engineering Research Centre of Translational Medicine and Innovative Drug, Changsha, ChinaDepartment of Pathology, Xiangya hospital and Department of Pathology, School of Basic Medicine, Central South University, Changsha, ChinaDepartment of Pharmacy, The Second Xiangya Hospital, Central South University, Changsha, ChinaDepartment of Dermatology, Hunan Engineering Research Center of Skin Health and Disease, Hunan Key Laboratory of Skin Cancer and Psoriasis, Xiangya Hospital, Central South University, Changsha, ChinaDepartment of Dermatology, Hunan Engineering Research Center of Skin Health and Disease, Hunan Key Laboratory of Skin Cancer and Psoriasis, Xiangya Hospital, Central South University, Changsha, ChinaDepartment of Medical Oncology, Harbin Medical University Cancer Hospital, Harbin, ChinaDepartment of Pharmacy, The Second Xiangya Hospital, Central South University, Changsha, ChinaDepartment of Dermatology, Hunan Engineering Research Center of Skin Health and Disease, Hunan Key Laboratory of Skin Cancer and Psoriasis, Xiangya Hospital, Central South University, Changsha, ChinaDepartment of Pharmacy, The Second Xiangya Hospital, Central South University, Changsha, ChinaDepartment of Pharmacy, School of Medicine, Hunan Normal University, Changsha, ChinaDepartment of Cancer Biology and Toxicology, Department of Pharmacology, College of Medicine, Markey Cancer Center, University of Kentucky, Lexington, Kentucky, USABackground Immune checkpoint blockade (ICB) targeting programmed death ligand-1 (PD-L1)/programmed cell death protein-1 (PD-1) pathway has become an attractive strategy for cancer treatment; however, unsatisfactory efficacy has limited its clinical benefits. Therefore, a more comprehensive understanding of the regulation of PD-L1 expression is essential for developing more effective cancer immunotherapy. Recent studies have revealed the important roles of eukaryotic elongation factor 2 kinase (eEF2K) in promoting epithelial-mesenchymal transition (EMT), angiogenesis, tumor cell migration and invasion; nevertheless, the exact role of eEF2K in the regulation of tumor immune microenvironment (TIME) remains largely unknown.Methods In this study, we used a cohort of 38 patients with melanoma who received anti-PD-1 treatment to explore the association between eEF2K expression and immunotherapy efficacy against melanoma. Immunoprecipitation-mass spectrometry analysis and in vitro assays were used to examine the role and molecular mechanism of eEF2K in regulating PD-L1 expression. We also determined the effects of eEF2K on tumor growth and cytotoxicity of CD8+ T cells in TIME in a mouse melanoma model. We further investigated the efficacy of the eEF2K inhibition in combination with anti-PD-1 treatment in vivo.Results High eEF2K expression is correlated with better therapeutic response and longer survival in patients with melanoma treated with PD-1 monoclonal antibody (mAb). Moreover, eEF2K protein expression is positively correlated with PD-L1 protein expression. Mechanistically, eEF2K directly bound to and inactivated glycogen synthase kinase 3 beta (GSK3β) by phosphorylating it at serine 9 (S9), leading to PD-L1 protein stabilization and upregulation, and subsequently tumor immune evasion. Knockdown of eEF2K decreased PD-L1 expression and enhanced CD8+ T cell activity, thus dramatically attenuating murine B16F10 melanoma growth in vivo. Clinically, p-GSK3β/S9 expression is positively correlated with the expressions of eEF2K and PD-L1, and the response to anti-PD-1 immunotherapy. Furthermore, eEF2K inhibitor, NH125 treatment or eEF2K knockdown enhanced the efficacy of PD-1 mAb therapy in a melanoma mouse model.Conclusions Our results suggest that eEF2K may serve as a biomarker for predicting therapeutic response and prognosis in patients receiving anti-PD-1 therapy, reveal a vital role of eEF2K in regulating TIME by controlling PD-L1 expression and provide a potential combination therapeutic strategy of eEF2K inhibition with ICB therapy.https://jitc.bmj.com/content/10/3/e004026.full
spellingShingle Jianxun Song
Xiang Chen
Ying Wen
Mingzhu Yin
Yan Cheng
Wenjun Yi
Xingcong Ren
Xisha Chen
Kuansong Wang
Shilong Jiang
Hongyin Sun
Xuanling Che
Minghui Zhang
Jiaying He
Mengting Liao
Xiangling Li
Xiaoming Zhou
Jinming Yang
eEF2K promotes PD-L1 stabilization through inactivating GSK3β in melanoma
Journal for ImmunoTherapy of Cancer
title eEF2K promotes PD-L1 stabilization through inactivating GSK3β in melanoma
title_full eEF2K promotes PD-L1 stabilization through inactivating GSK3β in melanoma
title_fullStr eEF2K promotes PD-L1 stabilization through inactivating GSK3β in melanoma
title_full_unstemmed eEF2K promotes PD-L1 stabilization through inactivating GSK3β in melanoma
title_short eEF2K promotes PD-L1 stabilization through inactivating GSK3β in melanoma
title_sort eef2k promotes pd l1 stabilization through inactivating gsk3β in melanoma
url https://jitc.bmj.com/content/10/3/e004026.full
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