{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T07:07:44Z","timestamp":1782284864735,"version":"3.54.5"},"reference-count":49,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,7,1]],"date-time":"2021-07-01T00:00:00Z","timestamp":1625097600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Oncol."],"abstract":"<jats:p>Malignant pleural mesothelioma is an aggressive cancer, heterogeneous in its presentation and behaviour. Despite an increasing knowledge about molecular markers and their diagnostic and prognostic value, they are not used as much as they might be for treatment allocation. It has been recently reported that mesothelioma cells that lack BAP1 (BRCA1 Associated Protein) are sensitive to inhibition of the EZH2 (Enhancer of Zeste Homolog 2) histone methyltransferase. Since we observed strong H3K27me3 (histone H3 lysine 27 trimetylation) immunoreactivity in BAP1 wild-type mesothelioma biopsies, we decided to characterize <jats:italic>in vitro<\/jats:italic> the response\/resistance of BAP1 wild-type mesothelioma cells to the EZH2 selective inhibitor, EPZ-6438. Here we demonstrate that BAP1 wild-type mesothelioma cells were rendered sensitive to EPZ-6438 upon SIRT1 (Sirtuin 1) silencing\/inhibition or when cultured as multicellular spheroids, in which SIRT1 expression was lower compared to cells grown in monolayers. Notably, treatment of spheroids with EPZ-6438 abolished H3K27me3 and induced the expression of <jats:italic>CDKN2A<\/jats:italic> (Cyclin-Dependent Kinase Inhibitor 2A), causing cell growth arrest. EPZ-6438 treatment also resulted in a rapid and sustained induction of the genes encoding HIF2\u03b1 (Hypoxia Inducible Factor 2\u03b1), TG2 (Transglutaminase 2) and IL-6 (Interleukin 6). Loss of <jats:italic>CDKN2<\/jats:italic> is a common event in mesothelioma. <jats:italic>CDKN2A<\/jats:italic> silencing in combination with EPZ-6438 treatment induced apoptotic death in mesothelioma spheroids. In a <jats:italic>CDKN2A<\/jats:italic> wild-type setting apoptosis was induced by combining EPZ-6438 with 1-155, a TG2 selective and irreversible inhibitor. In conclusion, our data suggests that the expression of <jats:italic>CDKN2A<\/jats:italic> predicts cell fate in response to EZH2 inhibition and could potentially stratify tumors likely to undergo apoptosis.<\/jats:p>","DOI":"10.3389\/fonc.2021.678447","type":"journal-article","created":{"date-parts":[[2021,7,1]],"date-time":"2021-07-01T17:24:39Z","timestamp":1625160279000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":16,"title":["CDKN2A Determines Mesothelioma Cell Fate to EZH2 Inhibition"],"prefix":"10.3389","volume":"11","author":[{"given":"Giulia","family":"Pinton","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhuo","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cecilia","family":"Balzano","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sara","family":"Missaglia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniela","family":"Tavian","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Renzo","family":"Boldorini","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dean A.","family":"Fennell","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martin","family":"Griffin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Laura","family":"Moro","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2021,7,1]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"2","DOI":"10.1111\/j.1440-1843.2005.00694.x","article-title":"Pathogenesis of Malignant Pleural Mesothelioma","volume":"10","author":"Jaurand","year":"2005","journal-title":"Respirology"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1038\/nrc.2017.42","article-title":"Novel Insights Into Mesothelioma Biology and Implications for Therapy","volume":"17","author":"Yap","year":"2017","journal-title":"Nat Rev Cancer"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1016\/j.aogh.2014.09.016","article-title":"The Global Spread of Asbestos","volume":"80","author":"Frank","year":"2014","journal-title":"Ann Glob Health"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-publhealth-031811-124704","article-title":"The Worldwide Pandemic of Asbestos-Related Diseases","volume":"34","author":"Stayner","year":"2013","journal-title":"Annu Rev Public Health"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1200\/JCO.2003.11.136","article-title":"Phase III Study of Pemetrexed in Combination With Cisplatin Versus Cisplatin Alone in Patients With Malignant Pleural Mesothelioma","volume":"21","author":"Vogelzang","year":"2003","journal-title":"J Clin Oncol"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1016\/j.athoracsur.2009.09.028","article-title":"Second Surgery for Recurrence of Malignant Pleural Mesothelioma After Extrapleural Pneumonectomy","volume":"89","author":"Politi","year":"2010","journal-title":"Ann Thorac Surg"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1016\/j.jtcvs.2014.10.128","article-title":"Updated Patterns of Failure After Multimodality Therapy for Malignant Pleural Mesothelioma","volume":"149","author":"Baldini","year":"2015","journal-title":"J Thorac Cardiovasc Surg"},{"key":"B8","doi-asserted-by":"publisher","first-page":"19","DOI":"10.3390\/ijms19061603","article-title":"Heterogeneity in Malignant Pleural Mesothelioma","volume":"30","author":"Oehl","year":"2018","journal-title":"Int J Mol Sci"},{"key":"B9","volume-title":"World Health Organization Classification of Tumours of the Lung, Pleura, Thymus and Heart","author":"Travis","year":"2015"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-14-1008","article-title":"Whole-Exome Sequencing Reveals Frequent Genetic Alterations in bap1, nf2, cdkn2a, and Cul1 in Malignant Pleural Mesothelioma","volume":"75","author":"Guo","year":"2015","journal-title":"Cancer Res"},{"key":"B11","doi-asserted-by":"publisher","first-page":"1333","DOI":"10.1038\/s41467-019-09307-6","article-title":"Dissecting Heterogeneity in Malignant Pleural Mesothelioma Through Histo-Molecular Gradients for Clinical Applications","volume":"10","author":"Blum","year":"2019","journal-title":"Nat Commun"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1158\/1535-7163.MCT-16-0229","article-title":"Genomic Landscape of Malignant Mesotheliomas","volume":"15","author":"Kato","year":"2016","journal-title":"Mol Cancer Ther"},{"key":"B13","doi-asserted-by":"publisher","first-page":"20","DOI":"10.3390\/ht7030020","article-title":"Genomics and Epigenetics of Malignant Mesothelioma","volume":"7","author":"Sage","year":"2018","journal-title":"High Throughput"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1038\/nrc.2017.42","article-title":"Novel Insight Into Mesothelioma Biology and Implications for Therapy","volume":"17","author":"Yap","year":"2017","journal-title":"Nat Rev Cancer"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1097\/JTO.0000000000000436","article-title":"Targeted Next-Generation Sequencing of Cancer Genes in Advanced Stage Malignant Pleural Mesothelioma: A Retrospective Study","volume":"10","author":"Lo Iacono","year":"2015","journal-title":"J Thorac Oncol"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1038\/ng.3520","article-title":"Comprehensive Genomic Analysis of Malignant Pleural Mesothelioma Identifies Recurrent Mutations, Gene Fusions and Splicing Alterations","volume":"48","author":"Bueno","year":"2016","journal-title":"Nat Genet"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1038\/nm.3947","article-title":"Loss of BAP1 Function Leads to EZH2-Dependent Transformation","volume":"21","author":"LaFave","year":"2015","journal-title":"Nat Med"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1126\/science.1118947","article-title":"Akt-Mediated Phosphorylation of EZH2 Suppresses Methylation of Lysine 27 in Histone H3","volume":"310","author":"Cha","year":"2005","journal-title":"Science"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkv238","article-title":"PCAF-Primed EZH2 Acetylation Regulates Its Stability and Promotes Lung Adenocarcinoma Progression","volume":"43","author":"Wan","year":"2015","journal-title":"Nucleic Acids Res"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1002\/emmm.201201783","article-title":"Smurf2-mediated Degradation of EZH2 Enhances Neuron Differentiation and Improves Functional Recovery After Ischaemic Stroke","volume":"5","author":"Yu","year":"2013","journal-title":"EMBO Mol Med"},{"key":"B21","doi-asserted-by":"publisher","first-page":"e2704","DOI":"10.1371\/journal.pone.0002704","article-title":"The Polycomb Repressive Complex 2 Is a Potential Target of SUMO Modifications","volume":"3","author":"Riising","year":"2008","journal-title":"PLoS One"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1323226111","article-title":"O-GlcNAcylation Regulates EZH2 Protein Stability and Function","volume":"111","author":"Chu","year":"2014","journal-title":"Proc Natl Acad Sci U S A"},{"key":"B23","article-title":"p16 Alterations and Deletion Mapping of 9p21-p22 in Malignant Mesothelioma","volume":"54","author":"Cheng","year":"1994","journal-title":"Cancer Res"},{"key":"B24","doi-asserted-by":"publisher","first-page":"507","DOI":"10.1186\/s12885-019-5652-y","article-title":"CDKN2A Copy Number and p16 Expression in Malignant Pleural\u00a0Mesothelioma\u00a0in Relation to Asbestos Exposure","volume":"19","author":"Kettunen","year":"2019","journal-title":"BMC Cancer"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-05-3907","article-title":"Global Gene Expression Profiling of Pleural Mesotheliomas: Overexpression of Aurora Kinases and P16\/CDKN2A Deletion as Prognostic Factors and Critical Evaluation of Microarray-Based Prognostic Prediction","volume":"66","author":"L\u00f3pez-R\u00edos","year":"2006","journal-title":"Cancer Res"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1007\/s00428-008-0689-3","article-title":"Prognostic Significance of p16\/cdkn2a Loss in Pleural Malignant Mesotheliomas","volume":"453","author":"Dacic","year":"2008","journal-title":"Virchows Arch"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1002\/ijc.23949","article-title":"Genomic Events Associated With Progression of Pleural Malignant Mesothelioma","volume":"124","author":"Ivanov","year":"2009","journal-title":"Int J Cancer"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1016\/S0169-5002(02)00178-2","article-title":"Inactivation of p16INK4a Expression in Malignant Mesothelioma by Methylation","volume":"38","author":"Wong","year":"2002","journal-title":"Lung Cancer"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1016\/j.biocel.2015.07.006","article-title":"JMJD3 as an Epigenetic Regulator in Development and Disease","volume":"67","author":"Burchfield","year":"2015","journal-title":"Int J Biochem Cell Biol"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1038\/nrd4154","article-title":"Histone Lysine Demethylases as Targets for Anticancer Therapy","volume":"12","author":"Hojfeldt","year":"2013","journal-title":"Nat Rev Drug Discov"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1016\/j.mrfmmm.2008.01.001","article-title":"Hypoxia Induces a Novel Signature of Chromatin Modifications and Global Repression of Transcription","volume":"640","author":"Johnson","year":"2008","journal-title":"Mutat Res"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.2217\/epi-2016-0025","article-title":"KDM6B Histone Demethylase Is an Epigenetic Regulator of Estrogen Receptor \u03b2 Expression in Human Pleural Mesothelioma","volume":"8","author":"Manente","year":"2016","journal-title":"Epigenomics"},{"key":"B33","doi-asserted-by":"publisher","first-page":"e2592","DOI":"10.1038\/cddis.2017.30","article-title":"Tissue Transglutaminase (TG2) Enables Survival of Human Malignant Pleural Mesothelioma Cells in Hypoxia","volume":"8","author":"Zonca","year":"2017","journal-title":"Cell Death Dis"},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.1038\/nrm1014","article-title":"Transglutaminases: Crosslinking Enzymes With Pleiotropic Functions","volume":"4","author":"Lorand","year":"2003","journal-title":"Nat Rev Mol Cell Biol"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.5625\/lar.2015.31.4.188","article-title":"Tissue Transglutaminase-Interleukin-6 Axis Facilitates Peritoneal Tumor Spreading and Metastasis of Human Ovarian Cancer Cells","volume":"31","author":"Oh","year":"2015","journal-title":"Lab Anim Res"},{"key":"B36","doi-asserted-by":"publisher","first-page":"724","DOI":"10.1186\/s12885-016-2746-7","article-title":"Il-1\u03b2 Induces IL-6 Production and Increases Invasiveness and Estrogen-Independent Growth in a TG2-dependent Manner in Human Breast Cancer Cells","volume":"16","author":"Oh","year":"2016","journal-title":"BMC Cancer"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2015.08.013","article-title":"Development of Potent and Selective Tissue Tranglutaminase Inhibitors: Their Effect on TG2 Function and Application in Pathological Conditions","volume":"22","author":"Badarau","year":"2015","journal-title":"Chem Biol"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1016\/j.bmcl.2008.09.006","article-title":"Synthesis of Potent Water-Soluble Tissue Transglutaminase Inhibitors","volume":"18","author":"Griffin","year":"2008","journal-title":"Bioorg Med Chem Lett"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.18632\/oncotarget.4486","article-title":"Intracellular Lactate-Mediated Induction of Estrogen Receptor Beta (Er\u03b2) in Biphasic Malignant Pleural Mesothelioma Cells","volume":"6","author":"Manente","year":"2015","journal-title":"Oncotarget"},{"key":"B40","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.pharmthera.2015.01.002","article-title":"Targeting Histone Lysine Methylation in Cancer","volume":"150","author":"McGrath","year":"2015","journal-title":"Pharmacol Ther"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1038\/nrc.2016.83","article-title":"Maintaining Cell Identity: PRC2-mediated Regulation of Transcription and Cancer","volume":"16","author":"Comet","year":"2016","journal-title":"Nat Rev Cancer"},{"key":"B42","doi-asserted-by":"publisher","first-page":"77","DOI":"10.1158\/1078-0432.CCR-11-0962","article-title":"Polycomb Repressor Complex-2 Is a Novel Target for Mesothelioma Therapy","volume":"18","author":"Kemp","year":"2012","journal-title":"Clin Cancer Res"},{"key":"B43","doi-asserted-by":"publisher","first-page":"8515","DOI":"10.1200\/JCO.2018.36.15_suppl.8515","article-title":"Phase 2, Multicenter Study of the EZH2 Inhibitor Tazemetostat as Monotherapy in Adults With Relapsed or Refractory (R\/R) Malignant Mesothelioma (MM) With BAP1 Inactivation","volume":"36","author":"Zauderer","year":"2018","journal-title":"J Clinic Oncol"},{"key":"B44","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1016\/j.ddtec.2017.03.002","article-title":"The Production of 3D Tumor Spheroids for Cancer Drug Discovery","volume":"23","author":"Sant","year":"2017","journal-title":"Drug Discov Today Technol"},{"key":"B45","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2010.05.023","article-title":"Sirtuin 1 Modulates Cellular Responses to Hypoxia by Deacetylating Hypoxia-Inducible Factor1alpha","volume":"38","author":"Lim","year":"2010","journal-title":"Mol Cell"},{"key":"B46","doi-asserted-by":"publisher","DOI":"10.1101\/gad.415507","article-title":"The Polycomb Group Proteins Bind Throughout the INK4A-ARF Locus and Are Disassociated in Senescent Cells","volume":"21","author":"Bracken","year":"2007","journal-title":"Genes Dev"},{"key":"B47","doi-asserted-by":"publisher","first-page":"94","DOI":"10.1038\/s41556-018-0249-2","article-title":"The Dynamic Nature of Senescence in Cancer","volume":"21","author":"Soyoung","year":"2019","journal-title":"Nat Cell Biol"},{"key":"B48","doi-asserted-by":"publisher","first-page":"55","DOI":"10.3978\/j.issn.2218-6751.2014.07.01","article-title":"The Role of Interleukin-6 in Malignant\u00a0Mesothelioma","volume":"4","author":"Abdul Rahim","year":"2015","journal-title":"Transl Lung Cancer Res"},{"key":"B49","doi-asserted-by":"publisher","DOI":"10.1038\/cdd.2009.162","article-title":"Polycomb Protein EZH2 Regulates E2F1-Dependent Apoptosis Through Epigenetically Modulating Bim Expression","volume":"17","author":"Wu","year":"2010","journal-title":"Cell Death Differ"}],"updated-by":[{"DOI":"10.3389\/fonc.2022.1081632","type":"corrigendum","label":"Corrigendum","source":"publisher","updated":{"date-parts":[[2022,11,23]],"date-time":"2022-11-23T00:00:00Z","timestamp":1669161600000}}],"container-title":["Frontiers in Oncology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fonc.2021.678447\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,11,22]],"date-time":"2022-11-22T14:57:20Z","timestamp":1669129040000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fonc.2021.678447\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,7,1]]},"references-count":49,"alternative-id":["10.3389\/fonc.2021.678447"],"URL":"https:\/\/doi.org\/10.3389\/fonc.2021.678447","relation":{"corrigendum":[{"id-type":"doi","id":"10.3389\/fonc.2022.1081632","asserted-by":"object"}]},"ISSN":["2234-943X"],"issn-type":[{"value":"2234-943X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,7,1]]},"article-number":"678447"}}