BRAF inhibitor resistance of melanoma cells triggers increased susceptibility to natural killer cell-mediated lysis

Background Targeted therapies and immunotherapies are first-line treatments for patients with advanced melanoma. Serine–threonine protein kinase B-RAF (BRAF) and mitogen-activated protein kinase (MEK) inhibition leads to a 70% response rate in patients with advanced melanoma with a BRAFV600E/K mutat...

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Main Authors: Antoine Toubert, Eric Pasmant, Alexandra Frazao, Louise Rethacker, Géraldine Jeudy, Marina Colombo, Marie-Françoise Avril, Helene Moins-Teisserenc, Marie Roelens, Sophie Dalac, Eve Maubec, Anne Caignard
Format: Article
Language:English
Published: BMJ Publishing Group 2020-10-01
Series:Journal for ImmunoTherapy of Cancer
Online Access:https://jitc.bmj.com/content/8/2/e000275.full
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author Antoine Toubert
Eric Pasmant
Alexandra Frazao
Louise Rethacker
Géraldine Jeudy
Marina Colombo
Marie-Françoise Avril
Helene Moins-Teisserenc
Marie Roelens
Sophie Dalac
Eve Maubec
Anne Caignard
author_facet Antoine Toubert
Eric Pasmant
Alexandra Frazao
Louise Rethacker
Géraldine Jeudy
Marina Colombo
Marie-Françoise Avril
Helene Moins-Teisserenc
Marie Roelens
Sophie Dalac
Eve Maubec
Anne Caignard
author_sort Antoine Toubert
collection DOAJ
description Background Targeted therapies and immunotherapies are first-line treatments for patients with advanced melanoma. Serine–threonine protein kinase B-RAF (BRAF) and mitogen-activated protein kinase (MEK) inhibition leads to a 70% response rate in patients with advanced melanoma with a BRAFV600E/K mutation. However, acquired resistance occurs in the majority of patients, leading to relapse. Immunotherapies that activate immune cytotoxic effectors induce long-lasting responses in 30% of patients. In that context, combination of targeted therapies with immunotherapy (IT) is a promising approach. We considered boosting natural killer (NK) cell tumor immunosurveillance, as melanoma cells express stress-induced molecules and activate NK cell lysis.Methods Here we have generated vemurafenib (a BRAF inihibitor)-resistant (R) cells from BRAFV600E SK28 and M14-sensitive (S) melanoma cell lines and investigated how resistance interferes with immunogenicity to NK cells. We determined the levels of several soluble molecules including NK ligands in 61 melanoma patients at baseline and 6 months M post-treatment with targeted therapies or immunotherapies.Results Vemurafenib resistance involved activation of p-AKT in SK28R and of p-MEK/p-ERK in M14R cells and was accompanied by modulation of NK ligands. Compared with S cells, SK28R displayed an increased expression of natural killer group 2 D (NKG2D) receptor ligands (major histocompatibility complex class (MHC) I chain-related protein A (MICA) and UL16-binding protein 2 (ULBP2)) whereas M14R exhibited decreased ULBP2 . SK28R and M14R cells induced higher NK degranulation and interferon gamma secretion and were more efficiently lysed by donor and patient NK cells. SK28R showed increased tumor necrosis factor-related apoptosis-inducing ligand receptor II (TRAIL-RII) expression and TRAIL-induced apoptosis, and TRAIL-induced apoptosis of M14R was decreased. Combined BRAF/MEK inhibitors abrogated the growth of SK28S, M14S, and M14R cells, while growth of SK28R was maintained. BRAF/MEK inhibition attenuated NK activity but R cell lines activated polyfunctional NK cells and were lysed with high efficiency. We investigated the relationship of soluble NK ligands and response to treatment in a series of melanoma patients. Soluble NKG2D ligands known to regulate the receptor function have been associated to cancer progression. Serum analysis of patients treated with target therapies or IT indicates that soluble forms of NK ligands (MICA, B7H6, programmed cell death ligand 1, and carcinoembryonic antigen cell adhesion molecule 1) may correlate with clinical response.Conclusion These results support strategies combining targeted therapies and NK-based immunotherapies.
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spelling doaj-art-6a710dde9f9d4917acf1c956f7e1ec9d2024-11-10T14:40:08ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262020-10-018210.1136/jitc-2019-000275BRAF inhibitor resistance of melanoma cells triggers increased susceptibility to natural killer cell-mediated lysisAntoine Toubert0Eric Pasmant1Alexandra Frazao2Louise Rethacker3Géraldine Jeudy4Marina Colombo5Marie-Françoise Avril6Helene Moins-Teisserenc7Marie Roelens8Sophie Dalac9Eve Maubec10Anne Caignard11Institut de Recherche Saint Louis, INSERM U1160, Université de Paris, Paris, France3 Department of Molecular Genetics, Hôpital Cochin, DMU BioPhyGen, AP-HP Centre-Université Paris Cité, Paris, FranceUniversité de Paris, INSERM UMRS-1160, Institut de Recherche Saint-Louis, 75010, Paris, FranceUniversité de Paris, INSERM UMRS-1160, Institut de Recherche Saint-Louis, 75010, Paris, FranceUniversity Hospital Centre Dijon Bocage Complex, Dermatology Department, Dijon, FranceUniversité de Paris, INSERM UMRS-1160, Institut de Recherche Saint-Louis, 75010, Paris, FranceUniversité de Paris, AP-HP Hôpital Cochin, Dermatology Department, Institute Cochin, Paris, FranceUniversité de Paris, INSERM UMRS-1160, AP-HP hopital Saint-Louis, Institut de Recherche Saint-Louis, 75010, Paris, FranceUniversité de Paris, INSERM UMRS-1160, AP-HP hopital Saint-Louis, Institut de Recherche Saint-Louis, 75010, Paris, FranceHospital Center University Dijon Bourgogne, Dijon, France13 Dermatology, Hopital Avicenne, Bobigny, FranceUniversité de Paris, INSERM UMRS-1160, Institut de Recherche Saint-Louis, 75010, Paris, FranceBackground Targeted therapies and immunotherapies are first-line treatments for patients with advanced melanoma. Serine–threonine protein kinase B-RAF (BRAF) and mitogen-activated protein kinase (MEK) inhibition leads to a 70% response rate in patients with advanced melanoma with a BRAFV600E/K mutation. However, acquired resistance occurs in the majority of patients, leading to relapse. Immunotherapies that activate immune cytotoxic effectors induce long-lasting responses in 30% of patients. In that context, combination of targeted therapies with immunotherapy (IT) is a promising approach. We considered boosting natural killer (NK) cell tumor immunosurveillance, as melanoma cells express stress-induced molecules and activate NK cell lysis.Methods Here we have generated vemurafenib (a BRAF inihibitor)-resistant (R) cells from BRAFV600E SK28 and M14-sensitive (S) melanoma cell lines and investigated how resistance interferes with immunogenicity to NK cells. We determined the levels of several soluble molecules including NK ligands in 61 melanoma patients at baseline and 6 months M post-treatment with targeted therapies or immunotherapies.Results Vemurafenib resistance involved activation of p-AKT in SK28R and of p-MEK/p-ERK in M14R cells and was accompanied by modulation of NK ligands. Compared with S cells, SK28R displayed an increased expression of natural killer group 2 D (NKG2D) receptor ligands (major histocompatibility complex class (MHC) I chain-related protein A (MICA) and UL16-binding protein 2 (ULBP2)) whereas M14R exhibited decreased ULBP2 . SK28R and M14R cells induced higher NK degranulation and interferon gamma secretion and were more efficiently lysed by donor and patient NK cells. SK28R showed increased tumor necrosis factor-related apoptosis-inducing ligand receptor II (TRAIL-RII) expression and TRAIL-induced apoptosis, and TRAIL-induced apoptosis of M14R was decreased. Combined BRAF/MEK inhibitors abrogated the growth of SK28S, M14S, and M14R cells, while growth of SK28R was maintained. BRAF/MEK inhibition attenuated NK activity but R cell lines activated polyfunctional NK cells and were lysed with high efficiency. We investigated the relationship of soluble NK ligands and response to treatment in a series of melanoma patients. Soluble NKG2D ligands known to regulate the receptor function have been associated to cancer progression. Serum analysis of patients treated with target therapies or IT indicates that soluble forms of NK ligands (MICA, B7H6, programmed cell death ligand 1, and carcinoembryonic antigen cell adhesion molecule 1) may correlate with clinical response.Conclusion These results support strategies combining targeted therapies and NK-based immunotherapies.https://jitc.bmj.com/content/8/2/e000275.full
spellingShingle Antoine Toubert
Eric Pasmant
Alexandra Frazao
Louise Rethacker
Géraldine Jeudy
Marina Colombo
Marie-Françoise Avril
Helene Moins-Teisserenc
Marie Roelens
Sophie Dalac
Eve Maubec
Anne Caignard
BRAF inhibitor resistance of melanoma cells triggers increased susceptibility to natural killer cell-mediated lysis
Journal for ImmunoTherapy of Cancer
title BRAF inhibitor resistance of melanoma cells triggers increased susceptibility to natural killer cell-mediated lysis
title_full BRAF inhibitor resistance of melanoma cells triggers increased susceptibility to natural killer cell-mediated lysis
title_fullStr BRAF inhibitor resistance of melanoma cells triggers increased susceptibility to natural killer cell-mediated lysis
title_full_unstemmed BRAF inhibitor resistance of melanoma cells triggers increased susceptibility to natural killer cell-mediated lysis
title_short BRAF inhibitor resistance of melanoma cells triggers increased susceptibility to natural killer cell-mediated lysis
title_sort braf inhibitor resistance of melanoma cells triggers increased susceptibility to natural killer cell mediated lysis
url https://jitc.bmj.com/content/8/2/e000275.full
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