Cargando…

Cancer cell-intrinsic expression of MHC II in lung cancer cell lines is actively restricted by MEK/ERK signaling and epigenetic mechanisms

BACKGROUND: Programmed death 1/programmed death ligand 1 (PD-1/PD-L1) targeted immunotherapy affords clinical benefit in ~20% of unselected patients with lung cancer. The factor(s) that determine whether a tumor responds or fails to respond to immunotherapy remains an active area of investigation. W...

Descripción completa

Detalles Bibliográficos
Autores principales: Neuwelt, Alexander J, Kimball, Abigail K, Johnson, Amber M, Arnold, Benjamin W, Bullock, Bonnie L, Kaspar, Rachael E, Kleczko, Emily K, Kwak, Jeff W, Wu, Meng-Han, Heasley, Lynn E, Doebele, Robert C, Li, Howard Y, Nemenoff, Raphael A, Clambey, Eric T
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BMJ Publishing Group 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7204826/
https://www.ncbi.nlm.nih.gov/pubmed/32312906
http://dx.doi.org/10.1136/jitc-2019-000441
_version_ 1783530128208297984
author Neuwelt, Alexander J
Kimball, Abigail K
Johnson, Amber M
Arnold, Benjamin W
Bullock, Bonnie L
Kaspar, Rachael E
Kleczko, Emily K
Kwak, Jeff W
Wu, Meng-Han
Heasley, Lynn E
Doebele, Robert C
Li, Howard Y
Nemenoff, Raphael A
Clambey, Eric T
author_facet Neuwelt, Alexander J
Kimball, Abigail K
Johnson, Amber M
Arnold, Benjamin W
Bullock, Bonnie L
Kaspar, Rachael E
Kleczko, Emily K
Kwak, Jeff W
Wu, Meng-Han
Heasley, Lynn E
Doebele, Robert C
Li, Howard Y
Nemenoff, Raphael A
Clambey, Eric T
author_sort Neuwelt, Alexander J
collection PubMed
description BACKGROUND: Programmed death 1/programmed death ligand 1 (PD-1/PD-L1) targeted immunotherapy affords clinical benefit in ~20% of unselected patients with lung cancer. The factor(s) that determine whether a tumor responds or fails to respond to immunotherapy remains an active area of investigation. We have previously defined divergent responsiveness of two KRAS-mutant cell lines to PD-1/PD-L1 blockade using an orthotopic, immunocompetent mouse model. Responsiveness to PD-1/PD-L1 checkpoint blockade correlates with an interferon gamma (IFNγ)-inducible gene signature and major histocompatibility complex class II (MHC II) expression by cancer cells. In the current study, we aim to identify therapeutic targets that can be manipulated in order to enhance cancer-cell-specific MHC II expression. METHODS: Responsiveness to IFNγ and induction of MHC II expression was assessed after various treatment conditions in mouse and human non-small cell lung cancer (NSCLC) cell lines using mass cytometric and flow cytometric analysis. RESULTS: Single-cell analysis using mass and flow cytometry demonstrated that IFNγ consistently induced PD-L1 and MHC class I (MHC I) across multiple murine and human NSCLC cell lines. In contrast, MHC II showed highly variable induction following IFNγ treatment both between lines and within lines. In mouse models of NSCLC, MHC II induction was inversely correlated with basal levels of phosphorylated extracellular signal-regulated kinase (ERK) 1/2, suggesting potential mitogen-activated protein (MAP) kinase-dependent antagonism of MHC II expression. To test this, cell lines were subjected to varying levels of stimulation with IFNγ, and assessed for MHC II expression in the presence or absence of mitogen-activated protein kinase kinase (MEK) inhibitors. IFNγ treatment in the presence of MEK inhibitors significantly enhanced MHC II induction across multiple lung cancer lines, with minimal impact on expression of either PD-L1 or MHC I. Inhibition of histone deacetylases (HDACs) also enhanced MHC II expression to a more modest extent. Combined MEK and HDAC inhibition led to greater MHC II expression than either treatment alone. CONCLUSIONS: These studies emphasize the active inhibitory role that epigenetic and ERK signaling cascades have in restricting cancer cell-intrinsic MHC II expression in NSCLC, and suggest that combinatorial blockade of these pathways may engender new responsiveness to checkpoint therapies.
format Online
Article
Text
id pubmed-7204826
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher BMJ Publishing Group
record_format MEDLINE/PubMed
spelling pubmed-72048262020-05-12 Cancer cell-intrinsic expression of MHC II in lung cancer cell lines is actively restricted by MEK/ERK signaling and epigenetic mechanisms Neuwelt, Alexander J Kimball, Abigail K Johnson, Amber M Arnold, Benjamin W Bullock, Bonnie L Kaspar, Rachael E Kleczko, Emily K Kwak, Jeff W Wu, Meng-Han Heasley, Lynn E Doebele, Robert C Li, Howard Y Nemenoff, Raphael A Clambey, Eric T J Immunother Cancer Basic Tumor Immunology BACKGROUND: Programmed death 1/programmed death ligand 1 (PD-1/PD-L1) targeted immunotherapy affords clinical benefit in ~20% of unselected patients with lung cancer. The factor(s) that determine whether a tumor responds or fails to respond to immunotherapy remains an active area of investigation. We have previously defined divergent responsiveness of two KRAS-mutant cell lines to PD-1/PD-L1 blockade using an orthotopic, immunocompetent mouse model. Responsiveness to PD-1/PD-L1 checkpoint blockade correlates with an interferon gamma (IFNγ)-inducible gene signature and major histocompatibility complex class II (MHC II) expression by cancer cells. In the current study, we aim to identify therapeutic targets that can be manipulated in order to enhance cancer-cell-specific MHC II expression. METHODS: Responsiveness to IFNγ and induction of MHC II expression was assessed after various treatment conditions in mouse and human non-small cell lung cancer (NSCLC) cell lines using mass cytometric and flow cytometric analysis. RESULTS: Single-cell analysis using mass and flow cytometry demonstrated that IFNγ consistently induced PD-L1 and MHC class I (MHC I) across multiple murine and human NSCLC cell lines. In contrast, MHC II showed highly variable induction following IFNγ treatment both between lines and within lines. In mouse models of NSCLC, MHC II induction was inversely correlated with basal levels of phosphorylated extracellular signal-regulated kinase (ERK) 1/2, suggesting potential mitogen-activated protein (MAP) kinase-dependent antagonism of MHC II expression. To test this, cell lines were subjected to varying levels of stimulation with IFNγ, and assessed for MHC II expression in the presence or absence of mitogen-activated protein kinase kinase (MEK) inhibitors. IFNγ treatment in the presence of MEK inhibitors significantly enhanced MHC II induction across multiple lung cancer lines, with minimal impact on expression of either PD-L1 or MHC I. Inhibition of histone deacetylases (HDACs) also enhanced MHC II expression to a more modest extent. Combined MEK and HDAC inhibition led to greater MHC II expression than either treatment alone. CONCLUSIONS: These studies emphasize the active inhibitory role that epigenetic and ERK signaling cascades have in restricting cancer cell-intrinsic MHC II expression in NSCLC, and suggest that combinatorial blockade of these pathways may engender new responsiveness to checkpoint therapies. BMJ Publishing Group 2020-04-19 /pmc/articles/PMC7204826/ /pubmed/32312906 http://dx.doi.org/10.1136/jitc-2019-000441 Text en © Author(s) (or their employer(s)) 2020. Re-use permitted under CC BY. Published by BMJ. https://creativecommons.org/licenses/by/4.0/This is an open access article distributed in accordance with the Creative Commons Attribution 4.0 Unported (CC BY 4.0) license, which permits others to copy, redistribute, remix, transform and build upon this work for any purpose, provided the original work is properly cited, a link to the licence is given, and indication of whether changes were made. See https://creativecommons.org/licenses/by/4.0/.
spellingShingle Basic Tumor Immunology
Neuwelt, Alexander J
Kimball, Abigail K
Johnson, Amber M
Arnold, Benjamin W
Bullock, Bonnie L
Kaspar, Rachael E
Kleczko, Emily K
Kwak, Jeff W
Wu, Meng-Han
Heasley, Lynn E
Doebele, Robert C
Li, Howard Y
Nemenoff, Raphael A
Clambey, Eric T
Cancer cell-intrinsic expression of MHC II in lung cancer cell lines is actively restricted by MEK/ERK signaling and epigenetic mechanisms
title Cancer cell-intrinsic expression of MHC II in lung cancer cell lines is actively restricted by MEK/ERK signaling and epigenetic mechanisms
title_full Cancer cell-intrinsic expression of MHC II in lung cancer cell lines is actively restricted by MEK/ERK signaling and epigenetic mechanisms
title_fullStr Cancer cell-intrinsic expression of MHC II in lung cancer cell lines is actively restricted by MEK/ERK signaling and epigenetic mechanisms
title_full_unstemmed Cancer cell-intrinsic expression of MHC II in lung cancer cell lines is actively restricted by MEK/ERK signaling and epigenetic mechanisms
title_short Cancer cell-intrinsic expression of MHC II in lung cancer cell lines is actively restricted by MEK/ERK signaling and epigenetic mechanisms
title_sort cancer cell-intrinsic expression of mhc ii in lung cancer cell lines is actively restricted by mek/erk signaling and epigenetic mechanisms
topic Basic Tumor Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7204826/
https://www.ncbi.nlm.nih.gov/pubmed/32312906
http://dx.doi.org/10.1136/jitc-2019-000441
work_keys_str_mv AT neuweltalexanderj cancercellintrinsicexpressionofmhciiinlungcancercelllinesisactivelyrestrictedbymekerksignalingandepigeneticmechanisms
AT kimballabigailk cancercellintrinsicexpressionofmhciiinlungcancercelllinesisactivelyrestrictedbymekerksignalingandepigeneticmechanisms
AT johnsonamberm cancercellintrinsicexpressionofmhciiinlungcancercelllinesisactivelyrestrictedbymekerksignalingandepigeneticmechanisms
AT arnoldbenjaminw cancercellintrinsicexpressionofmhciiinlungcancercelllinesisactivelyrestrictedbymekerksignalingandepigeneticmechanisms
AT bullockbonniel cancercellintrinsicexpressionofmhciiinlungcancercelllinesisactivelyrestrictedbymekerksignalingandepigeneticmechanisms
AT kasparrachaele cancercellintrinsicexpressionofmhciiinlungcancercelllinesisactivelyrestrictedbymekerksignalingandepigeneticmechanisms
AT kleczkoemilyk cancercellintrinsicexpressionofmhciiinlungcancercelllinesisactivelyrestrictedbymekerksignalingandepigeneticmechanisms
AT kwakjeffw cancercellintrinsicexpressionofmhciiinlungcancercelllinesisactivelyrestrictedbymekerksignalingandepigeneticmechanisms
AT wumenghan cancercellintrinsicexpressionofmhciiinlungcancercelllinesisactivelyrestrictedbymekerksignalingandepigeneticmechanisms
AT heasleylynne cancercellintrinsicexpressionofmhciiinlungcancercelllinesisactivelyrestrictedbymekerksignalingandepigeneticmechanisms
AT doebelerobertc cancercellintrinsicexpressionofmhciiinlungcancercelllinesisactivelyrestrictedbymekerksignalingandepigeneticmechanisms
AT lihowardy cancercellintrinsicexpressionofmhciiinlungcancercelllinesisactivelyrestrictedbymekerksignalingandepigeneticmechanisms
AT nemenoffraphaela cancercellintrinsicexpressionofmhciiinlungcancercelllinesisactivelyrestrictedbymekerksignalingandepigeneticmechanisms
AT clambeyerict cancercellintrinsicexpressionofmhciiinlungcancercelllinesisactivelyrestrictedbymekerksignalingandepigeneticmechanisms