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Nintedanib and Dasatinib Treatments Induce Protective Autophagy as a Potential Resistance Mechanism in MPM Cells

Malignant pleural mesothelioma (MPM) is a rare type of cancer with a grim prognosis. So far, no targetable oncogenic mutation was identified in MPM and biomarkers with predictive value toward drug sensitivity or resistance are also lacking. Nintedanib (BIBF1120) is a small-molecule tyrosine kinase i...

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Autores principales: Hegedüs, Luca, Szücs, Kata D., Kudla, Matthias, Heidenreich, Julian, Jendrossek, Verena, Peña-Llopis, Samuel, Garay, Tamas, Czirok, Andras, Aigner, Clemens, Plönes, Till, Vega-Rubin-de-Celis, Silvia, Hegedüs, Balazs
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8982261/
https://www.ncbi.nlm.nih.gov/pubmed/35392170
http://dx.doi.org/10.3389/fcell.2022.852812
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author Hegedüs, Luca
Szücs, Kata D.
Kudla, Matthias
Heidenreich, Julian
Jendrossek, Verena
Peña-Llopis, Samuel
Garay, Tamas
Czirok, Andras
Aigner, Clemens
Plönes, Till
Vega-Rubin-de-Celis, Silvia
Hegedüs, Balazs
author_facet Hegedüs, Luca
Szücs, Kata D.
Kudla, Matthias
Heidenreich, Julian
Jendrossek, Verena
Peña-Llopis, Samuel
Garay, Tamas
Czirok, Andras
Aigner, Clemens
Plönes, Till
Vega-Rubin-de-Celis, Silvia
Hegedüs, Balazs
author_sort Hegedüs, Luca
collection PubMed
description Malignant pleural mesothelioma (MPM) is a rare type of cancer with a grim prognosis. So far, no targetable oncogenic mutation was identified in MPM and biomarkers with predictive value toward drug sensitivity or resistance are also lacking. Nintedanib (BIBF1120) is a small-molecule tyrosine kinase inhibitor that showed promising efficacy preclinically and in phase II trial in MPM as an angiogenesis inhibitor combined with chemotherapy. However, the extended phase III trial failed. In this study, we investigated the effect of nintedanib on one of its targets, the SRC kinase, in two commercial and six novel MPM cell lines. Surprisingly, nintedanib treatment did not inhibit SRC activation in MPM cells and even increased phosphorylation of SRC in several cell lines. Combination treatment with the SRC inhibitor dasatinib could reverse this effect in all cell lines, however, the cellular response was dependent on the drug sensitivity of the cells. In 2 cell lines, with high sensitivity to both nintedanib and dasatinib, the drug combination had no synergistic effect but cell death was initiated. In 2 cell lines insensitive to nintedanib combination treatment reduced cell viability synergisticaly without cell death. In contrast, in these cells both treatments increased the autophagic flux assessed by degradation of the autophagy substrate p62 and increased presence of LC3B-II, increased number of GFP-LC3 puncta and decreased readings of the HiBiT-LC3 reporter. Additionaly, autophagy was synergistically promoted by the combined treatment. At the transcriptional level, analysis of lysosomal biogenesis regulator Transcription Factor EB (TFEB) showed that in all cell lines treated with nintedanib and to a lesser extent, with dasatinib, it became dephosphorylated and accumulated in the nucleus. Interestingly, the expression of certain known TFEB target genes implicated in autophagy or lysosomal biogenesis were significantly modified only in 1 cell line. Finally, we showed that autophagy induction in our MPM cell lines panel by nintedanib and dasatinib is independent of the AKT/mTOR and the ERK pathways. Our study reveals that autophagy can serve as a cytoprotective mechanism following nintedanib or dasatinib treatments in MPM cells.
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spelling pubmed-89822612022-04-06 Nintedanib and Dasatinib Treatments Induce Protective Autophagy as a Potential Resistance Mechanism in MPM Cells Hegedüs, Luca Szücs, Kata D. Kudla, Matthias Heidenreich, Julian Jendrossek, Verena Peña-Llopis, Samuel Garay, Tamas Czirok, Andras Aigner, Clemens Plönes, Till Vega-Rubin-de-Celis, Silvia Hegedüs, Balazs Front Cell Dev Biol Cell and Developmental Biology Malignant pleural mesothelioma (MPM) is a rare type of cancer with a grim prognosis. So far, no targetable oncogenic mutation was identified in MPM and biomarkers with predictive value toward drug sensitivity or resistance are also lacking. Nintedanib (BIBF1120) is a small-molecule tyrosine kinase inhibitor that showed promising efficacy preclinically and in phase II trial in MPM as an angiogenesis inhibitor combined with chemotherapy. However, the extended phase III trial failed. In this study, we investigated the effect of nintedanib on one of its targets, the SRC kinase, in two commercial and six novel MPM cell lines. Surprisingly, nintedanib treatment did not inhibit SRC activation in MPM cells and even increased phosphorylation of SRC in several cell lines. Combination treatment with the SRC inhibitor dasatinib could reverse this effect in all cell lines, however, the cellular response was dependent on the drug sensitivity of the cells. In 2 cell lines, with high sensitivity to both nintedanib and dasatinib, the drug combination had no synergistic effect but cell death was initiated. In 2 cell lines insensitive to nintedanib combination treatment reduced cell viability synergisticaly without cell death. In contrast, in these cells both treatments increased the autophagic flux assessed by degradation of the autophagy substrate p62 and increased presence of LC3B-II, increased number of GFP-LC3 puncta and decreased readings of the HiBiT-LC3 reporter. Additionaly, autophagy was synergistically promoted by the combined treatment. At the transcriptional level, analysis of lysosomal biogenesis regulator Transcription Factor EB (TFEB) showed that in all cell lines treated with nintedanib and to a lesser extent, with dasatinib, it became dephosphorylated and accumulated in the nucleus. Interestingly, the expression of certain known TFEB target genes implicated in autophagy or lysosomal biogenesis were significantly modified only in 1 cell line. Finally, we showed that autophagy induction in our MPM cell lines panel by nintedanib and dasatinib is independent of the AKT/mTOR and the ERK pathways. Our study reveals that autophagy can serve as a cytoprotective mechanism following nintedanib or dasatinib treatments in MPM cells. Frontiers Media S.A. 2022-03-22 /pmc/articles/PMC8982261/ /pubmed/35392170 http://dx.doi.org/10.3389/fcell.2022.852812 Text en Copyright © 2022 Hegedüs, Szücs, Kudla, Heidenreich, Jendrossek, Peña-Llopis, Garay, Czirok, Aigner, Plönes, Vega-Rubin-de-Celis and Hegedüs. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cell and Developmental Biology
Hegedüs, Luca
Szücs, Kata D.
Kudla, Matthias
Heidenreich, Julian
Jendrossek, Verena
Peña-Llopis, Samuel
Garay, Tamas
Czirok, Andras
Aigner, Clemens
Plönes, Till
Vega-Rubin-de-Celis, Silvia
Hegedüs, Balazs
Nintedanib and Dasatinib Treatments Induce Protective Autophagy as a Potential Resistance Mechanism in MPM Cells
title Nintedanib and Dasatinib Treatments Induce Protective Autophagy as a Potential Resistance Mechanism in MPM Cells
title_full Nintedanib and Dasatinib Treatments Induce Protective Autophagy as a Potential Resistance Mechanism in MPM Cells
title_fullStr Nintedanib and Dasatinib Treatments Induce Protective Autophagy as a Potential Resistance Mechanism in MPM Cells
title_full_unstemmed Nintedanib and Dasatinib Treatments Induce Protective Autophagy as a Potential Resistance Mechanism in MPM Cells
title_short Nintedanib and Dasatinib Treatments Induce Protective Autophagy as a Potential Resistance Mechanism in MPM Cells
title_sort nintedanib and dasatinib treatments induce protective autophagy as a potential resistance mechanism in mpm cells
topic Cell and Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8982261/
https://www.ncbi.nlm.nih.gov/pubmed/35392170
http://dx.doi.org/10.3389/fcell.2022.852812
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