N-terminal acetylation of transcription factor LIP induces immune therapy resistance via suppression of PD-L1 expression in non-small cell lung cancer

Background Programmed death-1 (PD-1) checkpoint blockade has revolutionized cancer therapy, yet its clinical success is confined to a subset of patients, underscoring the urgent need to understand the molecular underpinnings of programmed cell death ligand 1 (PD-L1) expression to combat immunotherap...

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Main Authors: Xing Gao, Xiang He, Yongshuo Liu, Feiyu Tang, Yuxi Tian, Siyuan Gong, Jia Shen, Aimin Wang, Lunquan Sun, Wensheng Wei, Liang Weng
Format: Article
Language:English
Published: BMJ Publishing Group 2024-11-01
Series:Journal for ImmunoTherapy of Cancer
Online Access:https://jitc.bmj.com/content/12/11/e009905.full
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author Xing Gao
Xiang He
Yongshuo Liu
Feiyu Tang
Yuxi Tian
Siyuan Gong
Jia Shen
Aimin Wang
Lunquan Sun
Wensheng Wei
Liang Weng
author_facet Xing Gao
Xiang He
Yongshuo Liu
Feiyu Tang
Yuxi Tian
Siyuan Gong
Jia Shen
Aimin Wang
Lunquan Sun
Wensheng Wei
Liang Weng
author_sort Xing Gao
collection DOAJ
description Background Programmed death-1 (PD-1) checkpoint blockade has revolutionized cancer therapy, yet its clinical success is confined to a subset of patients, underscoring the urgent need to understand the molecular underpinnings of programmed cell death ligand 1 (PD-L1) expression to combat immunotherapy resistance.Methods Employing CRISPR/Cas9 screening, we identified key regulators of PD-L1 in non-small cell lung cancer (NSCLC) cells, focusing on the transcription factor CEBPB and its isoform liver-enriched inhibitory protein (LIP). Through chromatin immunoprecipitation (ChIP) and luciferase reporter assays, we explored the interaction between LIP and basic-helix-loop-helix E22 (BHLHE22) in controlling PD-L1 transcription. We also used immunofluorescence and NBD-CI assays to examine how N-terminal acetylation affects LIP’s subcellular localization. The impact of LIP on tumor growth was assessed via subcutaneous tumorigenicity assays, while immunohistochemistry and immunofluorescence were used to analyze LIP-induced alterations in the tumor immune microenvironment.Results Our research indicates that CEBPB, particularly its LIP isoform, significantly suppresses PD-L1 expression in NSCLC cells. This suppression is contingent on LIP’s N-terminal acetylation by the N-terminal acetyltransferase A complex, which facilitates LIP’s nuclear entry and interaction with BHLHE22. This interaction leads to the formation of a co-repressor complex at the PD-L1 promoter, effectively reducing PD-L1 expression and enhancing the tumor immune response.Conclusions Identifying CEBPB, especially the LIP isoform, as a pivotal regulator of PD-L1 expression sheds light on the mechanisms behind PD-1 blockade resistance in NSCLC. Our findings suggest that modulating LIP’s function or its molecular interactions might offer a novel approach to boosting the efficacy of immunotherapies.
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series Journal for ImmunoTherapy of Cancer
spelling doaj-art-6bea76d1a64e4fdd9e9b1d5e40cd00142024-12-15T23:30:11ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262024-11-01121110.1136/jitc-2024-009905N-terminal acetylation of transcription factor LIP induces immune therapy resistance via suppression of PD-L1 expression in non-small cell lung cancerXing Gao0Xiang He1Yongshuo Liu2Feiyu Tang3Yuxi Tian4Siyuan Gong5Jia Shen6Aimin Wang7Lunquan Sun8Wensheng Wei9Liang Weng10Center of Oral and Maxillofacial Cancer (COMAC), Xiangya Hospital Central South University, Changsha, Hunan, ChinaKey Laboratory of Molecular Radiation Oncology Hunan Province, Xiangya Cancer Center, Xiangya Hospital Central South University, Changsha, ChinaDepartment of Pathology and Lab Medicine, Shandong Academy of Medical Sciences, Jinan, Shandong, ChinaCenter for Biotherapy, Sun Yat-Sen University, Guangzhou, ChinaDepartment of Geriatric Respiratory and Critical Care Medicine, Xiangya Hospital Central South University, Changsha, Hunan, ChinaKey Laboratory of Molecular Radiation Oncology Hunan Province, Xiangya Cancer Center, Xiangya Hospital Central South University, Changsha, ChinaNational Clinical Research Center for Geriatric Disorders, Xiangya Hospital Central South University, Changsha, ChinaDepartment of Emergency, Xiangya Hospital Central South University, Changsha, ChinaKey Laboratory of Molecular Radiation Oncology Hunan Province, Xiangya Cancer Center, Xiangya Hospital Central South University, Changsha, ChinaBiomedical Pioneering Innovation Center, Peking-Tsinghua Center for Life Sciences, Peking University Genome Editing Research Center, State Key Laboratory of Protein and Plant Gene Research, Peking University School of Life Sciences, Beijing, ChinaDepartment of Pathology, School of Basic Medical Sciences, Peking University Third Hospital, Peking University Health Science Center, Beijing, ChinaBackground Programmed death-1 (PD-1) checkpoint blockade has revolutionized cancer therapy, yet its clinical success is confined to a subset of patients, underscoring the urgent need to understand the molecular underpinnings of programmed cell death ligand 1 (PD-L1) expression to combat immunotherapy resistance.Methods Employing CRISPR/Cas9 screening, we identified key regulators of PD-L1 in non-small cell lung cancer (NSCLC) cells, focusing on the transcription factor CEBPB and its isoform liver-enriched inhibitory protein (LIP). Through chromatin immunoprecipitation (ChIP) and luciferase reporter assays, we explored the interaction between LIP and basic-helix-loop-helix E22 (BHLHE22) in controlling PD-L1 transcription. We also used immunofluorescence and NBD-CI assays to examine how N-terminal acetylation affects LIP’s subcellular localization. The impact of LIP on tumor growth was assessed via subcutaneous tumorigenicity assays, while immunohistochemistry and immunofluorescence were used to analyze LIP-induced alterations in the tumor immune microenvironment.Results Our research indicates that CEBPB, particularly its LIP isoform, significantly suppresses PD-L1 expression in NSCLC cells. This suppression is contingent on LIP’s N-terminal acetylation by the N-terminal acetyltransferase A complex, which facilitates LIP’s nuclear entry and interaction with BHLHE22. This interaction leads to the formation of a co-repressor complex at the PD-L1 promoter, effectively reducing PD-L1 expression and enhancing the tumor immune response.Conclusions Identifying CEBPB, especially the LIP isoform, as a pivotal regulator of PD-L1 expression sheds light on the mechanisms behind PD-1 blockade resistance in NSCLC. Our findings suggest that modulating LIP’s function or its molecular interactions might offer a novel approach to boosting the efficacy of immunotherapies.https://jitc.bmj.com/content/12/11/e009905.full
spellingShingle Xing Gao
Xiang He
Yongshuo Liu
Feiyu Tang
Yuxi Tian
Siyuan Gong
Jia Shen
Aimin Wang
Lunquan Sun
Wensheng Wei
Liang Weng
N-terminal acetylation of transcription factor LIP induces immune therapy resistance via suppression of PD-L1 expression in non-small cell lung cancer
Journal for ImmunoTherapy of Cancer
title N-terminal acetylation of transcription factor LIP induces immune therapy resistance via suppression of PD-L1 expression in non-small cell lung cancer
title_full N-terminal acetylation of transcription factor LIP induces immune therapy resistance via suppression of PD-L1 expression in non-small cell lung cancer
title_fullStr N-terminal acetylation of transcription factor LIP induces immune therapy resistance via suppression of PD-L1 expression in non-small cell lung cancer
title_full_unstemmed N-terminal acetylation of transcription factor LIP induces immune therapy resistance via suppression of PD-L1 expression in non-small cell lung cancer
title_short N-terminal acetylation of transcription factor LIP induces immune therapy resistance via suppression of PD-L1 expression in non-small cell lung cancer
title_sort n terminal acetylation of transcription factor lip induces immune therapy resistance via suppression of pd l1 expression in non small cell lung cancer
url https://jitc.bmj.com/content/12/11/e009905.full
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