ZEB1 transcription factor promotes immune escape in melanoma

Background The efficacy of immunotherapies in metastatic melanoma depends on a robust T cell infiltration. Oncogenic alterations of tumor cells have been associated to T cell exclusion. Identifying novel cancer cell-intrinsic non-genetic mechanisms of immune escape, the targeting of which would rein...

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Main Authors: Christophe Caux, Stephane Dalle, Arnaud de la Fouchardière, Laurie Tonon, Maud Plaschka, Valentin Benboubker, Maxime Grimont, Justine Berthet, Jonathan Lopez, Myrtille Le-Bouar, Brigitte Balme, Garance Tondeur, Lionel Larue, Alain Puisieux, Yenkel Grinberg-Bleyer, Nathalie Bendriss-Vermare, Bertrand Dubois, Julie Caramel
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/e003484.full
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author Christophe Caux
Stephane Dalle
Arnaud de la Fouchardière
Laurie Tonon
Maud Plaschka
Valentin Benboubker
Maxime Grimont
Justine Berthet
Jonathan Lopez
Myrtille Le-Bouar
Brigitte Balme
Garance Tondeur
Lionel Larue
Alain Puisieux
Yenkel Grinberg-Bleyer
Nathalie Bendriss-Vermare
Bertrand Dubois
Julie Caramel
author_facet Christophe Caux
Stephane Dalle
Arnaud de la Fouchardière
Laurie Tonon
Maud Plaschka
Valentin Benboubker
Maxime Grimont
Justine Berthet
Jonathan Lopez
Myrtille Le-Bouar
Brigitte Balme
Garance Tondeur
Lionel Larue
Alain Puisieux
Yenkel Grinberg-Bleyer
Nathalie Bendriss-Vermare
Bertrand Dubois
Julie Caramel
author_sort Christophe Caux
collection DOAJ
description Background The efficacy of immunotherapies in metastatic melanoma depends on a robust T cell infiltration. Oncogenic alterations of tumor cells have been associated to T cell exclusion. Identifying novel cancer cell-intrinsic non-genetic mechanisms of immune escape, the targeting of which would reinstate T cell recruitment, would allow to restore the response to anti-programmed cell death protein 1 (PD-1) antibody therapy. The epithelial-to-mesenchymal transition (EMT)-inducing transcription factor ZEB1 is a major regulator of melanoma cell plasticity, driving resistance to mitogen-activated protein kinase (MAPK) targeted therapies. We thus wondered whether ZEB1 signaling in melanoma cells may promote immune evasion and resistance to immunotherapy.Methods We evaluated the putative correlation between ZEB1 expression in melanoma cells and the composition of the immune infiltrate in a cohort of 60 human melanoma samples by combining transcriptomic (RNA-sequencing) and seven-color spatial multi-immunofluorescence analyses. Algorithm-based spatial reconstitution of tumors allowed the quantification of CD8+, CD4+ T cells number and their activation state (PD-1, Ki67). ZEB1 gain-of-function or loss-of-function approaches were then implemented in syngeneic melanoma mouse models, followed by monitoring of tumor growth, quantification of immune cell populations frequency and function by flow cytometry, cytokines secretion by multiplex analyses. Chromatin-immunoprecipitation was used to demonstrate the direct binding of this transcription factor on the promoters of cytokine-encoding genes. Finally, the sensitivity to anti-PD-1 antibody therapy upon ZEB1 gain-of-function or loss-of-function was evaluated.Results Combined spatial and transcriptomic analyses of the immune infiltrates in human melanoma samples demonstrated that ZEB1 expression in melanoma cells is associated with decreased CD8+ T cell infiltration, independently of β-catenin pathway activation. ZEB1 ectopic expression in melanoma cells impairs CD8+ T cell recruitment in syngeneic mouse models, resulting in tumor immune evasion and resistance to immune checkpoint blockade. Mechanistically, we demonstrate that ZEB1 directly represses the secretion of T cell-attracting chemokines, including CXCL10. Finally, Zeb1 knock-out, by promoting CD8+ T cell infiltration, synergizes with anti-PD-1 antibody therapy in promoting tumor regression.Conclusions We identify the ZEB1 transcription factor as a key determinant of melanoma immune escape, highlighting a previously unknown therapeutic target to increase efficacy of immunotherapy in melanoma.Trial registration number NCT02828202.
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spelling doaj-art-0e92f0aaa28a4889857d550e5e0db9eb2025-08-20T03:05:24ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262022-03-0110310.1136/jitc-2021-003484ZEB1 transcription factor promotes immune escape in melanomaChristophe Caux0Stephane Dalle1Arnaud de la Fouchardière2Laurie Tonon3Maud Plaschka4Valentin Benboubker5Maxime Grimont6Justine Berthet7Jonathan Lopez8Myrtille Le-Bouar9Brigitte Balme10Garance Tondeur11Lionel Larue12Alain Puisieux13Yenkel Grinberg-Bleyer14Nathalie Bendriss-Vermare15Bertrand Dubois16Julie Caramel17Laboratory of Immunotherapy of Cancer of Lyon (LICL), Lyon, France8 Hospices Civils de Lyon, Pierre Bénite, FranceCancer Cell Plasticity in Melanoma, Cancer Research Center of Lyon, Université de Lyon, Université Claude Bernard Lyon, INSERM 1052, CNRS 5286, Centre Léon Bérard, Lyon, FranceFondation Synergie Lyon Cancer, Plateforme de bio-informatique Gilles Thomas, Centre Léon Bérard, Lyon, FranceCancer Cell Plasticity in Melanoma, Cancer Research Center of Lyon, Université de Lyon, Université Claude Bernard Lyon, INSERM 1052, CNRS 5286, Centre Léon Bérard, Lyon, FranceCancer Cell Plasticity in Melanoma, Cancer Research Center of Lyon, Université de Lyon, Université Claude Bernard Lyon, INSERM 1052, CNRS 5286, Centre Léon Bérard, Lyon, FranceCancer Cell Plasticity in Melanoma, Cancer Research Center of Lyon, Université de Lyon, Université Claude Bernard Lyon, INSERM 1052, CNRS 5286, Centre Léon Bérard, Lyon, FranceCancer Immune Surveillance and Therapeutic Targeting, Cancer Research Center of Lyon, Université de Lyon, Université Claude Bernard Lyon 1, INSERM 1052, CNRS 5286, Centre Léon Bérard, Lyon, FranceCancer Cell Plasticity in Melanoma, Cancer Research Center of Lyon, Université de Lyon, Université Claude Bernard Lyon, INSERM 1052, CNRS 5286, Centre Léon Bérard, Lyon, FranceDermatology Unit, Hospices Civils de Lyon, CH Lyon Sud, Pierre Bénite, FranceDermatology Unit, Hospices Civils de Lyon, CH Lyon Sud, Pierre Bénite, FranceDermatology Unit, Hospices Civils de Lyon, CH Lyon Sud, Pierre Bénite, FranceInstitut Curie, PSL Research University, Paris, FranceInstitut Curie, PSL Research University, Paris, FranceUniversité Claude Bernard Lyon 1, Inserm U1052, CNRS 5286, Cancer Research Center of Lyon, Lyon, France4INSERM, Bordeaux, FranceCancer Immune Surveillance and Therapeutic Targeting, Cancer Research Center of Lyon, Université de Lyon, Université Claude Bernard Lyon 1, INSERM 1052, CNRS 5286, Centre Léon Bérard, Lyon, FranceCancer Cell Plasticity in Melanoma, Cancer Research Center of Lyon, Université de Lyon, Université Claude Bernard Lyon, INSERM 1052, CNRS 5286, Centre Léon Bérard, Lyon, FranceBackground The efficacy of immunotherapies in metastatic melanoma depends on a robust T cell infiltration. Oncogenic alterations of tumor cells have been associated to T cell exclusion. Identifying novel cancer cell-intrinsic non-genetic mechanisms of immune escape, the targeting of which would reinstate T cell recruitment, would allow to restore the response to anti-programmed cell death protein 1 (PD-1) antibody therapy. The epithelial-to-mesenchymal transition (EMT)-inducing transcription factor ZEB1 is a major regulator of melanoma cell plasticity, driving resistance to mitogen-activated protein kinase (MAPK) targeted therapies. We thus wondered whether ZEB1 signaling in melanoma cells may promote immune evasion and resistance to immunotherapy.Methods We evaluated the putative correlation between ZEB1 expression in melanoma cells and the composition of the immune infiltrate in a cohort of 60 human melanoma samples by combining transcriptomic (RNA-sequencing) and seven-color spatial multi-immunofluorescence analyses. Algorithm-based spatial reconstitution of tumors allowed the quantification of CD8+, CD4+ T cells number and their activation state (PD-1, Ki67). ZEB1 gain-of-function or loss-of-function approaches were then implemented in syngeneic melanoma mouse models, followed by monitoring of tumor growth, quantification of immune cell populations frequency and function by flow cytometry, cytokines secretion by multiplex analyses. Chromatin-immunoprecipitation was used to demonstrate the direct binding of this transcription factor on the promoters of cytokine-encoding genes. Finally, the sensitivity to anti-PD-1 antibody therapy upon ZEB1 gain-of-function or loss-of-function was evaluated.Results Combined spatial and transcriptomic analyses of the immune infiltrates in human melanoma samples demonstrated that ZEB1 expression in melanoma cells is associated with decreased CD8+ T cell infiltration, independently of β-catenin pathway activation. ZEB1 ectopic expression in melanoma cells impairs CD8+ T cell recruitment in syngeneic mouse models, resulting in tumor immune evasion and resistance to immune checkpoint blockade. Mechanistically, we demonstrate that ZEB1 directly represses the secretion of T cell-attracting chemokines, including CXCL10. Finally, Zeb1 knock-out, by promoting CD8+ T cell infiltration, synergizes with anti-PD-1 antibody therapy in promoting tumor regression.Conclusions We identify the ZEB1 transcription factor as a key determinant of melanoma immune escape, highlighting a previously unknown therapeutic target to increase efficacy of immunotherapy in melanoma.Trial registration number NCT02828202.https://jitc.bmj.com/content/10/3/e003484.full
spellingShingle Christophe Caux
Stephane Dalle
Arnaud de la Fouchardière
Laurie Tonon
Maud Plaschka
Valentin Benboubker
Maxime Grimont
Justine Berthet
Jonathan Lopez
Myrtille Le-Bouar
Brigitte Balme
Garance Tondeur
Lionel Larue
Alain Puisieux
Yenkel Grinberg-Bleyer
Nathalie Bendriss-Vermare
Bertrand Dubois
Julie Caramel
ZEB1 transcription factor promotes immune escape in melanoma
Journal for ImmunoTherapy of Cancer
title ZEB1 transcription factor promotes immune escape in melanoma
title_full ZEB1 transcription factor promotes immune escape in melanoma
title_fullStr ZEB1 transcription factor promotes immune escape in melanoma
title_full_unstemmed ZEB1 transcription factor promotes immune escape in melanoma
title_short ZEB1 transcription factor promotes immune escape in melanoma
title_sort zeb1 transcription factor promotes immune escape in melanoma
url https://jitc.bmj.com/content/10/3/e003484.full
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