{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,24]],"date-time":"2026-01-24T09:04:34Z","timestamp":1769245474824,"version":"3.49.0"},"reference-count":75,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2019,12,14]],"date-time":"2019-12-14T00:00:00Z","timestamp":1576281600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/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":["POCI-01-0145-FEDER-29043"],"award-info":[{"award-number":["POCI-01-0145-FEDER-29043"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["SFRH\/BD\/132751\/2017"],"award-info":[{"award-number":["SFRH\/BD\/132751\/2017"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004622","name":"KWF Kankerbestrijding","doi-asserted-by":"publisher","award":["Grant 13-6001"],"award-info":[{"award-number":["Grant 13-6001"]}],"id":[{"id":"10.13039\/501100004622","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Liquid biopsy-based biomarkers, such as microRNAs, represent valuable tools for patient management, but often do not make it to integration in the clinic. We aim to explore issues impeding this transition, in the setting of germ cell tumors, for which novel biomarkers are needed. We describe a model for identifying and validating clinically relevant microRNAs for germ cell tumor patients, using both in vitro, in vivo (mouse model) and patient-derived data. Initial wide screening of candidate microRNAs is performed, followed by targeted profiling of potentially relevant biomarkers. We demonstrate the relevance of appropriate (negative) controls, experimental conditions (proliferation), and issues related to sample origin (serum, plasma, cerebral spinal fluid) and pre-analytical variables (hemolysis, contaminants, temperature), all of which could interfere with liquid biopsy-based studies and their conclusions. Finally, we show the value of our identification model in a specific scenario, contradicting the presumed role of miR-375 as marker of teratoma histology in liquid biopsy setting. Our findings indicate other putative microRNAs (miR-885-5p, miR-448 and miR-197-3p) fulfilling this clinical need. The identification model is informative to identify the best candidate microRNAs to pursue in a clinical setting.<\/jats:p>","DOI":"10.3390\/cells8121637","type":"journal-article","created":{"date-parts":[[2019,12,16]],"date-time":"2019-12-16T05:19:38Z","timestamp":1576473578000},"page":"1637","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":90,"title":["Identification and Validation Model for Informative Liquid Biopsy-Based microRNA Biomarkers: Insights from Germ Cell Tumor In Vitro, In Vivo and Patient-Derived Data"],"prefix":"10.3390","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6829-1391","authenticated-orcid":false,"given":"Jo\u00e3o","family":"Lobo","sequence":"first","affiliation":[{"name":"Princess M\u00e1xima Center for Pediatric Oncology, Heidelberglaan 25, 3584 CS Utrecht, The Netherlands"},{"name":"Department of Pathology, Portuguese Oncology Institute of Porto (IPOP), R. Dr. Ant\u00f3nio Bernardino de Almeida, 4200-072 Porto, Portugal"},{"name":"Cancer Biology and Epigenetics Group, Research Center of Portuguese Oncology Institute of Porto (GEBC CI-IPOP) and Porto Comprehensive Cancer Center (P.CCC), R. Dr. Ant\u00f3nio Bernardino de Almeida, 4200-072 Porto, Portugal"},{"name":"Department of Pathology and Molecular Immunology, Institute of Biomedical Sciences Abel Salazar, University of Porto (ICBAS-UP), Rua Jorge Viterbo Ferreira 228, 4050-513 Porto, Portugal"}]},{"given":"Ad J.M.","family":"Gillis","sequence":"additional","affiliation":[{"name":"Princess M\u00e1xima Center for Pediatric Oncology, Heidelberglaan 25, 3584 CS Utrecht, The Netherlands"}]},{"given":"Annette","family":"van den Berg","sequence":"additional","affiliation":[{"name":"Princess M\u00e1xima Center for Pediatric Oncology, Heidelberglaan 25, 3584 CS Utrecht, The Netherlands"}]},{"given":"Lambert C.J.","family":"Dorssers","sequence":"additional","affiliation":[{"name":"Department of Pathology, Lab. for Exp. Patho-Oncology (LEPO), Erasmus MC-University Medical Center Rotterdam, Cancer Institute, Be-432A, PO Box 2040, 3000 CA Rotterdam, The Netherlands"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8613-3450","authenticated-orcid":false,"given":"Gafanzer","family":"Belge","sequence":"additional","affiliation":[{"name":"University Bremen, Faculty of Biology &amp; Chemistry, Bibliothekstra\u00dfe 1, 28359 Bremen, Germany"}]},{"given":"Klaus-Peter","family":"Dieckmann","sequence":"additional","affiliation":[{"name":"Department of Urology, Hodentumorzentrum Hamburg, Asklepios Klinik Altona, Hamburg, Germany &amp; Department of Urology, Albertinen Krankenhaus, Paul Ehrlich Strasse 1, 22763 Hamburg, Germany"}]},{"given":"Henk P.","family":"Roest","sequence":"additional","affiliation":[{"name":"Department of Surgery, Erasmus MC-University Medical Center Rotterdam, Cancer Institute, Be-432A, PO Box 2040, 3000 CA Rotterdam, The Netherlands"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0651-5334","authenticated-orcid":false,"given":"Luc J.W.","family":"van der Laan","sequence":"additional","affiliation":[{"name":"Department of Surgery, Erasmus MC-University Medical Center Rotterdam, Cancer Institute, Be-432A, PO Box 2040, 3000 CA Rotterdam, The Netherlands"}]},{"given":"Jourik","family":"Gietema","sequence":"additional","affiliation":[{"name":"University of Groningen, University Medical Center Groningen, Department of Medical Oncology, Hanzeplein 1, 9713 GZ Groningen, The Netherlands"}]},{"given":"Robert J.","family":"Hamilton","sequence":"additional","affiliation":[{"name":"Division of Urology, Department of Surgery, Princess Margaret Cancer Centre, University Health Network, 610 University Ave., 3-130, Toronto, ON M5G 2M9, Canada"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4186-5345","authenticated-orcid":false,"given":"Carmen","family":"Jer\u00f3nimo","sequence":"additional","affiliation":[{"name":"Cancer Biology and Epigenetics Group, Research Center of Portuguese Oncology Institute of Porto (GEBC CI-IPOP) and Porto Comprehensive Cancer Center (P.CCC), R. Dr. Ant\u00f3nio Bernardino de Almeida, 4200-072 Porto, Portugal"},{"name":"Department of Pathology and Molecular Immunology, Institute of Biomedical Sciences Abel Salazar, University of Porto (ICBAS-UP), Rua Jorge Viterbo Ferreira 228, 4050-513 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3171-4666","authenticated-orcid":false,"given":"Rui","family":"Henrique","sequence":"additional","affiliation":[{"name":"Department of Pathology, Portuguese Oncology Institute of Porto (IPOP), R. Dr. Ant\u00f3nio Bernardino de Almeida, 4200-072 Porto, Portugal"},{"name":"Cancer Biology and Epigenetics Group, Research Center of Portuguese Oncology Institute of Porto (GEBC CI-IPOP) and Porto Comprehensive Cancer Center (P.CCC), R. Dr. Ant\u00f3nio Bernardino de Almeida, 4200-072 Porto, Portugal"},{"name":"Department of Pathology and Molecular Immunology, Institute of Biomedical Sciences Abel Salazar, University of Porto (ICBAS-UP), Rua Jorge Viterbo Ferreira 228, 4050-513 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9956-7116","authenticated-orcid":false,"given":"Daniela","family":"Salvatori","sequence":"additional","affiliation":[{"name":"Central Laboratory Animal Facility, Leiden University Medical Center, Einthovenweg 20, Leiden 2333 ZC, The Netherlands"},{"name":"Department of Pathobiology, Anatomy and Physiology Division, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3584 CL Utrecht, The Netherlands"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8146-1911","authenticated-orcid":false,"given":"Leendert H.J.","family":"Looijenga","sequence":"additional","affiliation":[{"name":"Princess M\u00e1xima Center for Pediatric Oncology, Heidelberglaan 25, 3584 CS Utrecht, The Netherlands"},{"name":"Department of Pathology, Lab. for Exp. Patho-Oncology (LEPO), Erasmus MC-University Medical Center Rotterdam, Cancer Institute, Be-432A, PO Box 2040, 3000 CA Rotterdam, The Netherlands"}]}],"member":"1968","published-online":{"date-parts":[[2019,12,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2912","DOI":"10.18632\/oncotarget.23131","article-title":"The dawn of the liquid biopsy in the fight against cancer","volume":"9","author":"Wang","year":"2018","journal-title":"Oncotarget"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1387\/ijdb.130020re","article-title":"Role of stem cell proteins and microRNAs in embryogenesis and germ cell cancer","volume":"57","author":"Eini","year":"2013","journal-title":"Int. J. Dev. Biol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"541","DOI":"10.1038\/s41571-018-0035-x","article-title":"Clinical utility of circulating non-coding RNAs\u2014An update","volume":"15","author":"Anfossi","year":"2018","journal-title":"Nat. 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