{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,8]],"date-time":"2026-01-08T06:43:51Z","timestamp":1767854631133,"version":"3.49.0"},"reference-count":23,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2021,5,18]],"date-time":"2021-05-18T00:00:00Z","timestamp":1621296000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100007368","name":"Fondazione Pisa","doi-asserted-by":"publisher","award":["153\/16"],"award-info":[{"award-number":["153\/16"]}],"id":[{"id":"10.13039\/100007368","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancers"],"abstract":"<jats:p>Background: Malignant pleural mesothelioma (MPM) is a fatal tumor with a poor prognosis. The recent developments of liquid biopsies could provide novel diagnostic and prognostic tools in oncology. However, there is limited information about the feasibility of this technique for MPMs. Here, we investigate whether cancer-specific DNA sequences can be detected in pleural fluids and plasma of MPM patients as free circulating tumor DNA (ctDNA). Methods: We performed whole-exome sequencing on 14 tumor biopsies from 14 patients, and we analyzed 20 patient-specific somatic mutations with digital droplet PCR (ddPCR) in pleural fluids and plasma, using them as cancer-specific tumor biomarkers. Results: Most of the selected mutations could be detected in pleural fluids (94%) and, noteworthy, in plasma (83%) with the use of ddPCR. Pleural fluids showed similar levels of somatically mutated ctDNA (median = 12.75%, average = 16.3%, standard deviation = 12.3) as those detected in solid biopsies (median = 21.95%; average = 22.21%; standard deviation = 9.57), and their paired difference was weakly statistically significant (p = 0.048). On the other hand, the paired difference between solid biopsies and ctDNA from plasma (median = 0.29%, average = 0.89%, standard deviation = 1.40) was highly statistically significant (p = 2.5 \u00d7 10\u22127), corresponding to the important drop of circulating somatically mutated DNA in the bloodstream. However, despite the tiny amount of ctDNA in plasma, varying from 5.57% down to 0.14%, the mutations were detectable at rates similar to those possible for other tumors. Conclusions: We found robust evidence that mutated DNA is spilled from MPMs, mostly into pleural fluids, proving the concept that liquid biopsies are feasible for MPM patients.<\/jats:p>","DOI":"10.3390\/cancers13102445","type":"journal-article","created":{"date-parts":[[2021,5,18]],"date-time":"2021-05-18T12:17:16Z","timestamp":1621340236000},"page":"2445","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Liquid Biopsies from Pleural Effusions and Plasma from Patients with Malignant Pleural Mesothelioma: A Feasibility Study"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8605-578X","authenticated-orcid":false,"given":"Gabriele","family":"Moretti","sequence":"first","affiliation":[{"name":"Department of Biology, Genetic Unit, University of Pisa, via Derna 1, 56126 Pisa, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1075-4319","authenticated-orcid":false,"given":"Paolo","family":"Aretini","sequence":"additional","affiliation":[{"name":"Fondazione Pisana per la Scienza, Via Ferruccio Giovannini 13, 56017 San Giuliano Terme, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Francesca","family":"Lessi","sequence":"additional","affiliation":[{"name":"Fondazione Pisana per la Scienza, Via Ferruccio Giovannini 13, 56017 San Giuliano Terme, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chiara Maria","family":"Mazzanti","sequence":"additional","affiliation":[{"name":"Fondazione Pisana per la Scienza, Via Ferruccio Giovannini 13, 56017 San Giuliano Terme, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8849-193X","authenticated-orcid":false,"given":"Guntulu","family":"Ak","sequence":"additional","affiliation":[{"name":"Eskisehir Osmangazi University Lung and Pleural Cancers Research and Clinical Center, Eskisehir 26000, Turkey"},{"name":"Department of Chest Diseases, Medical Faculty, Eskisehir Osmangazi University, Eskisehir 26000, Turkey"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4812-0170","authenticated-orcid":false,"given":"Muzaffer","family":"Metinta\u015f","sequence":"additional","affiliation":[{"name":"Eskisehir Osmangazi University Lung and Pleural Cancers Research and Clinical Center, Eskisehir 26000, Turkey"},{"name":"Department of Chest Diseases, Medical Faculty, Eskisehir Osmangazi University, Eskisehir 26000, Turkey"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cecilia","family":"Lando","sequence":"additional","affiliation":[{"name":"IRCCS Ospedale Policlinico San Martino, Clinical Epidemiology, 16132 Genova, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4682-393X","authenticated-orcid":false,"given":"Rosa Angela","family":"Filiberti","sequence":"additional","affiliation":[{"name":"IRCCS Ospedale Policlinico San Martino, Clinical Epidemiology, 16132 Genova, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marco","family":"Lucchi","sequence":"additional","affiliation":[{"name":"Division of Thoracic Surgery, Cardiac-Thoracic and Vascular Department, University Hospital of Pisa, 56124 Pisa, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alessandra","family":"Bonotti","sequence":"additional","affiliation":[{"name":"Preventive and Occupational Medicine, University Hospital of Pisa, 56126 Pisa, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rudy","family":"Foddis","sequence":"additional","affiliation":[{"name":"Department of Translational Research and of New Technologies in Medicine and Surgery, University of Pisa, 56126 Pisa, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6575-5701","authenticated-orcid":false,"given":"Alfonso","family":"Cristaudo","sequence":"additional","affiliation":[{"name":"Department of Translational Research and of New Technologies in Medicine and Surgery, University of Pisa, 56126 Pisa, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5343-7271","authenticated-orcid":false,"given":"Andrea","family":"Bottari","sequence":"additional","affiliation":[{"name":"Department of Biology, Genetic Unit, University of Pisa, via Derna 1, 56126 Pisa, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alessandro","family":"Apollo","sequence":"additional","affiliation":[{"name":"Department of Biology, Genetic Unit, University of Pisa, via Derna 1, 56126 Pisa, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7343-6161","authenticated-orcid":false,"given":"Marzia","family":"Del Re","sequence":"additional","affiliation":[{"name":"Division of Pharmacology, Department of Internal Medicine, University of Pisa, 55, Via Roma, 56126 Pisa, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4414-8934","authenticated-orcid":false,"given":"Romano","family":"Danesi","sequence":"additional","affiliation":[{"name":"Division of Pharmacology, Department of Internal Medicine, University of Pisa, 55, Via Roma, 56126 Pisa, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1578-2637","authenticated-orcid":false,"given":"Luciano","family":"Mutti","sequence":"additional","affiliation":[{"name":"Sbarro Institute for Cancer Research and Molecular Medicine, Center for Biotechnology, College of Science and Technology, Temple University, Philadelphia, PA 19104, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Federica","family":"Gemignani","sequence":"additional","affiliation":[{"name":"Department of Biology, Genetic Unit, University of Pisa, via Derna 1, 56126 Pisa, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8364-6357","authenticated-orcid":false,"given":"Stefano","family":"Landi","sequence":"additional","affiliation":[{"name":"Department of Biology, Genetic Unit, University of Pisa, via Derna 1, 56126 Pisa, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,5,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"587","DOI":"10.1097\/JTO.0b013e31823f45c1","article-title":"Inflammation-based prognostic indices in malignant pleural mesothelioma","volume":"7","author":"Pinato","year":"2012","journal-title":"J. Thorac. Oncol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"25543","DOI":"10.1073\/pnas.2007622117","article-title":"Asbestos induces mesothelial cell transformation via HMGB1-driven autophagy","volume":"117","author":"Xue","year":"2020","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"598","DOI":"10.1158\/1078-0432.CCR-11-2259","article-title":"Molecular pathways: Targeting mechanisms of asbestos and erionite carcinogenesis in mesothelioma","volume":"18","author":"Carbone","year":"2012","journal-title":"Clin. Cancer Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2645","DOI":"10.1038\/s41388-018-0153-z","article-title":"How asbestos drives the tissue towards tumors: YAP activation, macrophage and mesothelial precursor recruitment, RNA editing, and somatic mutations","volume":"37","author":"Rehrauer","year":"2018","journal-title":"Oncogene"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2722","DOI":"10.1016\/j.ejca.2007.09.018","article-title":"Italian Mesothelioma Register (ReNaM) Working Group Analysis of latency time and its determinants in asbestos related malignant mesothelioma cases of the Italian register","volume":"43","author":"Marinaccio","year":"2007","journal-title":"Eur. J. Cancer"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"402","DOI":"10.3322\/caac.21572","article-title":"Mesothelioma: Scientific clues for prevention, diagnosis, and therapy","volume":"69","author":"Carbone","year":"2019","journal-title":"CA Cancer J. Clin."},{"key":"ref_7","first-page":"134","article-title":"Diagnostic Imaging and workup of Malignant Pleural Mesothelioma","volume":"88","author":"Cardinale","year":"2017","journal-title":"Acta Bio-Medica Atenei Parm."},{"key":"ref_8","first-page":"182","article-title":"Advances in the diagnosis, treatment and prognosis of malignant pleural mesothelioma","volume":"3","author":"Zhang","year":"2015","journal-title":"Ann. Transl. Med."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"426","DOI":"10.1038\/nrc3066","article-title":"Cell-free nucleic acids as biomarkers in cancer patients","volume":"11","author":"Schwarzenbach","year":"2011","journal-title":"Nat. Rev. Cancer"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1007\/978-3-319-22909-6_10","article-title":"Circulating Tumor Cells","volume":"Volume 882","author":"Stearns","year":"2016","journal-title":"Novel Biomarkers in the Continuum of Breast Cancer"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"869","DOI":"10.1080\/14737140.2019.1670063","article-title":"Liquid biopsies in pancreatic cancer","volume":"19","author":"Kamyabi","year":"2019","journal-title":"Expert Rev. Anticancer Ther."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1248","DOI":"10.1016\/j.jtho.2018.05.030","article-title":"Liquid Biopsy for Advanced Non-Small Cell Lung Cancer (NSCLC): A Statement Paper from the IASLC","volume":"13","author":"Rolfo","year":"2018","journal-title":"J. Thorac. Oncol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"114","DOI":"10.1186\/s12943-019-1043-x","article-title":"Liquid biopsy in hepatocellular carcinoma: Circulating tumor cells and circulating tumor DNA","volume":"18","author":"Ye","year":"2019","journal-title":"Mol. Cancer"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1007\/s10549-019-05365-y","article-title":"Overexpression of TK1 and CDK9 in plasma-derived exosomes is associated with clinical resistance to CDK4\/6 inhibitors in metastatic breast cancer patients","volume":"178","author":"Bertolini","year":"2019","journal-title":"Breast. Cancer Res. Treat."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1500","DOI":"10.1158\/1078-0432.CCR-17-1181","article-title":"ESR1 Methylation: A Liquid Biopsy-Based Epigenetic Assay for the Follow-up of Patients with Metastatic Breast Cancer Receiving Endocrine Treatment","volume":"24","author":"Mastoraki","year":"2018","journal-title":"Clin. Cancer Res."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Del Re, M., Crucitta, S., Gianfilippo, G., Passaro, A., Petrini, I., Restante, G., Michelucci, A., Fogli, S., de Marinis, F., and Porta, C. (2019). Understanding the Mechanisms of Resistance in EGFR-Positive NSCLC: From Tissue to Liquid Biopsy to Guide Treatment Strategy. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20163951"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Saenz-Anto\u00f1anzas, A., Auzmendi-Iriarte, J., Carrasco-Garcia, E., Moreno-Cugnon, L., Ruiz, I., Villanua, J., Ega\u00f1a, L., Otaegui, D., Sampr\u00f3n, N., and Matheu, A. (2019). Liquid Biopsy in Glioblastoma: Opportunities, Applications and Challenges. Cancers, 11.","DOI":"10.3390\/cancers11070950"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Sriram, K.B., Relan, V., Clarke, B.E., Duhig, E.E., Windsor, M.N., Matar, K.S., Naidoo, R., Passmore, L., McCaul, E., and Courtney, D. (2012). Pleural fluid cell-free DNA integrity index to identify cytologically negative malignant pleural effusions including mesotheliomas. BMC Cancer, 12.","DOI":"10.1186\/1471-2407-12-428"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/j.lungcan.2018.07.028","article-title":"Tumor-specific genetic variants can be detected in circulating cell-free DNA of malignant pleural mesothelioma patients","volume":"124","author":"Hylebos","year":"2018","journal-title":"Lung Cancer"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"589","DOI":"10.1093\/bioinformatics\/btp698","article-title":"Fast and accurate long-read alignment with Burrows-Wheeler transform","volume":"26","author":"Li","year":"2010","journal-title":"Bioinforma Oxf. Engl."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"568","DOI":"10.1101\/gr.129684.111","article-title":"VarScan 2: Somatic mutation and copy number alteration discovery in cancer by exome sequencing","volume":"22","author":"Koboldt","year":"2012","journal-title":"Genome Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"491","DOI":"10.1038\/ng.806","article-title":"A framework for variation discovery and genotyping using next-generation DNA sequencing data","volume":"43","author":"DePristo","year":"2011","journal-title":"Nat Genet."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1742","DOI":"10.3389\/fonc.2020.01742","article-title":"The Use of Immunohistochemistry, Fluorescence in situ Hybridization, and Emerging Epigenetic Markers in the Diagnosis of Malignant Pleural Mesothelioma (MPM): A Review","volume":"10","author":"Rozitis","year":"2020","journal-title":"Front. Oncol."}],"container-title":["Cancers"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6694\/13\/10\/2445\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:03:22Z","timestamp":1760162602000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6694\/13\/10\/2445"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,5,18]]},"references-count":23,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2021,5]]}},"alternative-id":["cancers13102445"],"URL":"https:\/\/doi.org\/10.3390\/cancers13102445","relation":{},"ISSN":["2072-6694"],"issn-type":[{"value":"2072-6694","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,5,18]]}}}