{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T22:04:22Z","timestamp":1789596262513,"version":"build-2803163510"},"reference-count":49,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2025,10,28]],"date-time":"2025-10-28T00:00:00Z","timestamp":1761609600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["2022.09566.BD"],"award-info":[{"award-number":["2022.09566.BD"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["pathsocjournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The Journal of Pathology"],"published-print":{"date-parts":[[2026,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Approximately 10% of testicular Leydig cell tumors (LCTs) are clinically malignant and unresponsive to systemic treatment. Predicting their clinical behavior can be problematic because there are no biomarkers that can consistently discriminate between benign and malignant LCTs. We assessed microRNA\u00a0expression profiles of LCTs to identify differentially expressed microRNAs that could potentially distinguish benign from malignant neoplasms. The study consisted of two phases. In the first (discovery) phase, we interrogated 768 microRNAs in a series of 11 LCTs (six malignant and five benign) using Taqman Low\u2010Density Array (TLDA) microRNA profiling. In the second phase, we validated the top differentially expressed microRNA targets with real\u2010time quantitative PCR on a series of 35 LCTs (17 malignant and 18 benign), assessing their clinical performance for distinguishing malignant from benign LCTs. Target biologic pathways were\u00a0analyzed using the miRTargetLink 2.0 tool. A total of 50 microRNAs were differentially regulated in malignant LCTs (27 upregulated, 23 downregulated). The top six microRNA candidates (top three upregulated and top three downregulated) were validated, showing good performance for discriminating between malignant and benign LCTs, with an area under the curve (AUC) ranging between 0.69 and 0.87. MiR\u2010196b\u20105p showed the best performance, with sensitivity, specificity, negative predictive value, positive predictive value, and accuracy of 82%, 83%, 83%, 82%, and 83%, respectively. A panel (i.e. combined) analysis reached 100% sensitivity and 83% specificity. Pathway analysis revealed significant overlap in the biological process targeted by the upregulated microRNAs in malignant LCTs, including proliferation, development, metabolism, hormone synthesis, and cell death. Our results support the idea that malignant LCTs are associated with a distinct microRNA signature. MiR\u2010196b\u20105p was identified as a potentially useful biomarker to distinguish benign from malignant tumors. The shared downstream targets of the top upregulated microRNAs suggest that dysregulation of cell proliferation and apoptosis underlie aggressive biologic behavior in LCTs and may offer opportunities for targeted therapies. \u00a9 2025 The Author(s).\n                    <jats:italic>The Journal of Pathology<\/jats:italic>\n                    published by John Wiley &amp; Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.\n                  <\/jats:p>","DOI":"10.1002\/path.6487","type":"journal-article","created":{"date-parts":[[2025,10,29]],"date-time":"2025-10-29T04:10:18Z","timestamp":1761711018000},"page":"54-64","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["<scp>MicroRNA<\/scp>\n                    profiling of testicular Leydig cell tumors identifies a\n                    <scp>microRNA<\/scp>\n                    signature associated with malignancy and\n                    <scp>miR<\/scp>\n                    \u2010196b\u20105p as a potentially useful biomarker"],"prefix":"10.1002","volume":"268","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6829-1391","authenticated-orcid":false,"given":"Jo\u00e3o","family":"Lobo","sequence":"first","affiliation":[{"name":"Department of Pathology Portuguese Oncology Institute of Porto (IPO Porto)\/Porto Comprehensive Cancer Center Raquel Seruca (P.CCC), R. Dr. Ant\u00f3nio Bernardino de Almeida  Porto Portugal"},{"name":"Cancer Biology and Epigenetics Group, IPO Porto Research Center (GEBC CI\u2010IPOP), Portuguese Oncology Institute of Porto (IPO Porto)\/Porto Comprehensive Cancer Center Raquel Seruca (P.CCC) &amp; CI\u2010IPOP@RISE (Health Research Network), R. Dr. Ant\u00f3nio Bernardino de Almeida  Porto Portugal"},{"name":"Department of Pathology and Molecular Immunology ICBAS \u2010 School of Medicine and Biomedical Sciences University of Porto  Porto Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nuno Tiago","family":"Tavares","sequence":"additional","affiliation":[{"name":"Cancer Biology and Epigenetics Group, IPO Porto Research Center (GEBC CI\u2010IPOP), Portuguese Oncology Institute of Porto (IPO Porto)\/Porto Comprehensive Cancer Center Raquel Seruca (P.CCC) &amp; CI\u2010IPOP@RISE (Health Research Network), R. Dr. Ant\u00f3nio Bernardino de Almeida  Porto Portugal"},{"name":"Doctoral Programme in Biomedical Sciences ICBAS \u2010 School of Medicine and Biomedical Sciences University of Porto  Porto Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-3931-0443","authenticated-orcid":false,"given":"Fernanda","family":"Fernandes\u2010Pontes","sequence":"additional","affiliation":[{"name":"Cancer Biology and Epigenetics Group, IPO Porto Research Center (GEBC CI\u2010IPOP), Portuguese Oncology Institute of Porto (IPO Porto)\/Porto Comprehensive Cancer Center Raquel Seruca (P.CCC) &amp; CI\u2010IPOP@RISE (Health Research Network), R. Dr. Ant\u00f3nio Bernardino de Almeida  Porto Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vera","family":"Const\u00e2ncio","sequence":"additional","affiliation":[{"name":"Cancer Biology and Epigenetics Group, IPO Porto Research Center (GEBC CI\u2010IPOP), Portuguese Oncology Institute of Porto (IPO Porto)\/Porto Comprehensive Cancer Center Raquel Seruca (P.CCC) &amp; CI\u2010IPOP@RISE (Health Research Network), R. Dr. Ant\u00f3nio Bernardino de Almeida  Porto Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana Teixeira","family":"Marques","sequence":"additional","affiliation":[{"name":"Cancer Biology and Epigenetics Group, IPO Porto Research Center (GEBC CI\u2010IPOP), Portuguese Oncology Institute of Porto (IPO Porto)\/Porto Comprehensive Cancer Center Raquel Seruca (P.CCC) &amp; CI\u2010IPOP@RISE (Health Research Network), R. Dr. Ant\u00f3nio Bernardino de Almeida  Porto Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bruno","family":"Oliveira\u2010Lopes","sequence":"additional","affiliation":[{"name":"Cancer Biology and Epigenetics Group, IPO Porto Research Center (GEBC CI\u2010IPOP), Portuguese Oncology Institute of Porto (IPO Porto)\/Porto Comprehensive Cancer Center Raquel Seruca (P.CCC) &amp; CI\u2010IPOP@RISE (Health Research Network), R. Dr. Ant\u00f3nio Bernardino de Almeida  Porto Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Diana","family":"Fonseca","sequence":"additional","affiliation":[{"name":"Cancer Biology and Epigenetics Group, IPO Porto Research Center (GEBC CI\u2010IPOP), Portuguese Oncology Institute of Porto (IPO Porto)\/Porto Comprehensive Cancer Center Raquel Seruca (P.CCC) &amp; CI\u2010IPOP@RISE (Health Research Network), R. Dr. Ant\u00f3nio Bernardino de Almeida  Porto Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carmen","family":"Jer\u00f3nimo","sequence":"additional","affiliation":[{"name":"Cancer Biology and Epigenetics Group, IPO Porto Research Center (GEBC CI\u2010IPOP), Portuguese Oncology Institute of Porto (IPO Porto)\/Porto Comprehensive Cancer Center Raquel Seruca (P.CCC) &amp; CI\u2010IPOP@RISE (Health Research Network), R. Dr. Ant\u00f3nio Bernardino de Almeida  Porto Portugal"},{"name":"Department of Pathology and Molecular Immunology ICBAS \u2010 School of Medicine and Biomedical Sciences University of Porto  Porto Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rui","family":"Henrique","sequence":"additional","affiliation":[{"name":"Department of Pathology Portuguese Oncology Institute of Porto (IPO Porto)\/Porto Comprehensive Cancer Center Raquel Seruca (P.CCC), R. Dr. Ant\u00f3nio Bernardino de Almeida  Porto Portugal"},{"name":"Cancer Biology and Epigenetics Group, IPO Porto Research Center (GEBC CI\u2010IPOP), Portuguese Oncology Institute of Porto (IPO Porto)\/Porto Comprehensive Cancer Center Raquel Seruca (P.CCC) &amp; CI\u2010IPOP@RISE (Health Research Network), R. Dr. Ant\u00f3nio Bernardino de Almeida  Porto Portugal"},{"name":"Department of Pathology and Molecular Immunology ICBAS \u2010 School of Medicine and Biomedical Sciences University of Porto  Porto Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3231-6870","authenticated-orcid":false,"given":"Kvetoslava","family":"Michalova","sequence":"additional","affiliation":[{"name":"Department of Pathology Charles University, Faculty of Medicine in Plzen, Bioptical Laboratory, Ltd  Pilsen Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kristine M","family":"Cornejo","sequence":"additional","affiliation":[{"name":"Department of Pathology Massachusetts General Hospital, Harvard Medical School  Boston MA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maurizio","family":"Colecchia","sequence":"additional","affiliation":[{"name":"Department of Pathology Vita\u2010Salute San Raffaele University, IRCCS San Raffaele Scientific Institute  Milan Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7254-4195","authenticated-orcid":false,"given":"Costantino","family":"Ricci","sequence":"additional","affiliation":[{"name":"Pathology Unit, DIAP\u2010Dipartimento Interaziendale di anatomia patologica di Bologna Maggiore Hospital\u2010AUSL Bologna  Bologna Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Muhammad T","family":"Idrees","sequence":"additional","affiliation":[{"name":"Department of Pathology Indiana University  Indianapolis IN USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Felix","family":"Contreras","sequence":"additional","affiliation":[{"name":"Laboratorio de Patolog\u00eda Cl\u00ednica Universitaria Uni\u00f3n M\u00e9dica, PUCMM  Santiago Dominican Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isabel M Fernandez","family":"Gonzalez","sequence":"additional","affiliation":[{"name":"Laboratorio de Patolog\u00eda Cl\u00ednica Universitaria Uni\u00f3n M\u00e9dica, PUCMM  Santiago Dominican Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William J","family":"Anderson","sequence":"additional","affiliation":[{"name":"Department of Pathology Brigham and Women's Hospital and Harvard Medical School  Boston MA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fiona","family":"MacLean","sequence":"additional","affiliation":[{"name":"Douglass Hanly Moir Pathology  Macquarie Park NSW Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adeboye O","family":"Osunkoya","sequence":"additional","affiliation":[{"name":"Department of Pathology and Urology Emory University School of Medicine  Atlanta GA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chia\u2010Sui","family":"Kao","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine Cleveland Clinic Foundation  Cleveland OH USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ankur R","family":"Sangoi","sequence":"additional","affiliation":[{"name":"Department of Pathology Stanford University  Stanford CA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas M","family":"Ulbright","sequence":"additional","affiliation":[{"name":"Department of Pathology Indiana University  Indianapolis IN USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8817-6331","authenticated-orcid":false,"given":"Andres M","family":"Acosta","sequence":"additional","affiliation":[{"name":"Department of Pathology Indiana University  Indianapolis IN USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,10,28]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00345-020-03079-1"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.urology.2014.10.021"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.urology.2019.09.016"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.urology.2019.02.014"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.juro.2014.01.104"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1002\/cncr.11573"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.eururo.2008.08.022"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1111\/his.14225"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1097\/JU.0000000000000705"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1097\/00000478-198503000-00002"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1442-2042.2010.02616.x"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1097\/00000478-199811000-00006"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1111\/his.14859"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41379-021-00845-3"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.euf.2019.05.012"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.clgc.2021.02.002"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.modpat.2023.100307"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1470-2045(12)70073-6"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2006.02.037"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41585-020-0296-x"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.path.2024.08.003"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1111\/his.14929"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.3390\/cells8121637"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1006\/meth.2001.1262"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0294236"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.humpath.2024.05.005"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1002\/path.6412"},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.1098\/rstb.2017.0338"},{"key":"e_1_2_8_30_1","doi-asserted-by":"publisher","DOI":"10.1002\/bimj.200410133"},{"key":"e_1_2_8_31_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkab297"},{"key":"e_1_2_8_32_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12964-025-02250-8"},{"key":"e_1_2_8_33_1","doi-asserted-by":"publisher","DOI":"10.5858\/2007-131-311-AILALC"},{"key":"e_1_2_8_34_1","doi-asserted-by":"publisher","DOI":"10.1097\/00000478-200211000-00004"},{"key":"e_1_2_8_35_1","doi-asserted-by":"publisher","DOI":"10.1097\/PAP.0000000000000423"},{"key":"e_1_2_8_36_1","first-page":"585","article-title":"Leydig cell tumors of the testis: a molecular\u2010cytogenetic study based on a large series of patients","volume":"17","author":"Verdorfer I","year":"2007","journal-title":"Oncol Rep"},{"key":"e_1_2_8_37_1","doi-asserted-by":"crossref","DOI":"10.1111\/his.15482","article-title":"Assessment and classification of sex cord\u2010stromal tumours of the testis: recommendations from the testicular sex cord\u2010stromal tumour (TESST) group, an expert panel of the genitourinary pathology society (GUPS) and International Society of Urological Pathology (ISUP)","author":"Acosta AM","year":"2025","journal-title":"Histopathology"},{"key":"e_1_2_8_38_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41576-023-00611-y"},{"key":"e_1_2_8_39_1","doi-asserted-by":"publisher","DOI":"10.1038\/d41586-024-03212-9"},{"key":"e_1_2_8_40_1","doi-asserted-by":"publisher","DOI":"10.1111\/andr.13258"},{"key":"e_1_2_8_41_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00432-025-06101-4"},{"key":"e_1_2_8_42_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00432-020-03429-x"},{"key":"e_1_2_8_43_1","doi-asserted-by":"publisher","DOI":"10.3390\/cancers12061601"},{"key":"e_1_2_8_44_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41416-023-02412-w"},{"key":"e_1_2_8_45_1","doi-asserted-by":"publisher","DOI":"10.1159\/000200077"},{"key":"e_1_2_8_46_1","doi-asserted-by":"publisher","DOI":"10.1093\/biolre\/ioy059"},{"key":"e_1_2_8_47_1","doi-asserted-by":"publisher","DOI":"10.1136\/gutjnl-2015-311256"},{"key":"e_1_2_8_48_1","doi-asserted-by":"publisher","DOI":"10.18632\/oncotarget.13580"},{"key":"e_1_2_8_49_1","doi-asserted-by":"publisher","DOI":"10.1002\/hep.22989"},{"key":"e_1_2_8_50_1","doi-asserted-by":"publisher","DOI":"10.1111\/his.15175"}],"container-title":["The Journal of Pathology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pathsocjournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/path.6487","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,12]],"date-time":"2025-12-12T07:41:42Z","timestamp":1765525302000},"score":1,"resource":{"primary":{"URL":"https:\/\/pathsocjournals.onlinelibrary.wiley.com\/doi\/10.1002\/path.6487"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,28]]},"references-count":49,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2026,1]]}},"alternative-id":["10.1002\/path.6487"],"URL":"https:\/\/doi.org\/10.1002\/path.6487","archive":["Portico"],"relation":{},"ISSN":["0022-3417","1096-9896"],"issn-type":[{"value":"0022-3417","type":"print"},{"value":"1096-9896","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,10,28]]},"assertion":[{"value":"2025-05-03","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-09-18","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-10-28","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}