{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:37:08Z","timestamp":1787027828965,"version":"build-2736575974"},"reference-count":46,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2026,4,16]],"date-time":"2026-04-16T00:00:00Z","timestamp":1776297600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000090","name":"Congressionally Directed Medical Research Programs","doi-asserted-by":"publisher","award":["W81XWH2210656"],"award-info":[{"award-number":["W81XWH2210656"]}],"id":[{"id":"10.13039\/100000090","id-type":"DOI","asserted-by":"publisher"}]},{"award":["W81XWH2210656"],"award-info":[{"award-number":["W81XWH2210656"]}],"id":[{"id":"https:\/\/ror.org\/03g2zjp07","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Background: High-grade serous ovarian carcinoma (HGSOC) is the most common and lethal subtype of epithelial ovarian cancer and is frequently diagnosed at advanced stages. Because currently available blood-based biomarkers have limited performance in early-stage disease, there is a need to identify circulating biomarker candidates associated with early-stage HGSOC. In this retrospective multi-institutional case\u2013control study, we evaluated whether serum extracellular vesicle (EV)-associated protein signatures distinguish early-stage HGSOC from healthy controls. Methods: Serum samples (n = 252) were obtained retrospectively from multiple institutions and included healthy controls and patients with early- and advanced-stage HGSOC. EV-associated proteins were profiled using liquid chromatography\u2013tandem mass spectrometry (LC\u2013MS\/MS) and proximity extension assay (PEA) to identify candidate proteins enriched in early-stage HGSOC. Selected candidates were evaluated by enzyme-linked immunosorbent assay (ELISA), and tissue-level expression was examined in early-stage HGSOC specimens. A multimarker combination model was generated using a smoothed empirical estimate of hyper-volume under the manifold (SHUM) approach and internally assessed by leave-one-out cross-validation. Results: Ten EV-associated serum proteins were prioritized on the basis of differential expression and fold change and were confirmed to be expressed in early-stage HGSOC tissues. In ELISA-based analyses, the combined 10-protein EV panel distinguished early-stage HGSOC from healthy controls with an area under the curve (AUC) of 0.99 in the study dataset, whereas MUC16 (CA-125) showed substantially lower performance in this comparison. The SHUM-based model yielded a true-positive rate of 0.971, a false-positive rate of 0.057, and a Matthews correlation coefficient of 0.915 in the analyzed cohort. Several candidate proteins were differentially enriched in EV fractions but not in matched whole serum. Conclusions: Serum EV-associated proteins are altered in early-stage HGSOC and define a multi-protein signature associated with this disease state in a retrospective case\u2013control setting. These findings support further evaluation of EV-based biomarker candidates in clinically representative and prospectively collected cohorts that include benign gynecologic conditions, symptomatic patients, and pre-diagnostic samples.<\/jats:p>","DOI":"10.3390\/cells15080706","type":"journal-article","created":{"date-parts":[[2026,4,16]],"date-time":"2026-04-16T11:35:42Z","timestamp":1776339342000},"page":"706","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Serum Extracellular Vesicle Protein Signatures Associated with Early-Stage High-Grade Serous Ovarian Carcinoma"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3535-2167","authenticated-orcid":false,"given":"Michelle","family":"Lightfoot","sequence":"first","affiliation":[{"name":"Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, NYU Langone Health\/Perlmutter Cancer Center, New York, NY 10016, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kalpana Deepa Priya","family":"Dorayappan","sequence":"additional","affiliation":[{"name":"Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, The Ohio State University Wexner Medical Center, Columbus, OH 43210, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vignesh","family":"Vudatha","sequence":"additional","affiliation":[{"name":"Department of Surgery, Virginia Commonwealth University School of Medicine, 1200 E Broad St., Richmond, VA 23298, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lakshmi Narasimhan.","family":"Chakrapani","sequence":"additional","affiliation":[{"name":"Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, The Ohio State University Wexner Medical Center, Columbus, OH 43210, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Priyam","family":"Das","sequence":"additional","affiliation":[{"name":"Department of Biostatistics, Virginia Commonwealth University, Richmond, VA 23298, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lianbo","family":"Yu","sequence":"additional","affiliation":[{"name":"Department of Biomedical Informatics, The Ohio State University Wexner Medical Center, Columbus, OH 43210, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Colin","family":"Hisey","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, The Ohio State University, Columbus, OH 43210, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takahiko","family":"Sakaue","sequence":"additional","affiliation":[{"name":"Division of Gastroenterology, Department of Medicine, Kurume University School of Medicine, 67 Asahi-machi, Kurume 830-0011, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thangavel","family":"Muthusamy","sequence":"additional","affiliation":[{"name":"Cellular and Molecular Biochemistry, Sree Balaji Medical College and Hospital, Bharath Institute of Higher Education and Research, Chennai 600073, Tamil Nadu, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Parthiban","family":"Panneerselvam","sequence":"additional","affiliation":[{"name":"University of The Cumberlands, Williamsburg, KY 40769, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Floor","family":"Backes","sequence":"additional","affiliation":[{"name":"Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, The Ohio State University Wexner Medical Center, Columbus, OH 43210, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Casey","family":"Cosgrove","sequence":"additional","affiliation":[{"name":"Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, The Ohio State University Wexner Medical Center, Columbus, OH 43210, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Derek","family":"Hansford","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, The Ohio State University, Columbus, OH 43210, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David E.","family":"Cohn","sequence":"additional","affiliation":[{"name":"Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, The Ohio State University Wexner Medical Center, Columbus, OH 43210, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David M.","family":"O\u2019Malley","sequence":"additional","affiliation":[{"name":"Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, The Ohio State University Wexner Medical Center, Columbus, OH 43210, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rajan","family":"Gogna","sequence":"additional","affiliation":[{"name":"Department of Surgery, Virginia Commonwealth University School of Medicine, 1200 E Broad St., Richmond, VA 23298, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8016-9134","authenticated-orcid":false,"given":"Karuppaiyah","family":"Selvendiran","sequence":"additional","affiliation":[{"name":"Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, The Ohio State University Wexner Medical Center, Columbus, OH 43210, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1038\/nrclinonc.2013.5","article-title":"Latest research and treatment of advanced-stage epithelial ovarian cancer","volume":"10","author":"Coleman","year":"2013","journal-title":"Nat. Rev. Clin. Oncol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"16061","DOI":"10.1038\/nrdp.2016.61","article-title":"Ovarian cancer","volume":"2","author":"Matulonis","year":"2016","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_3","first-page":"10","article-title":"Cancer statistics, 2025","volume":"75","author":"Siegel","year":"2025","journal-title":"CA Cancer J Clin"},{"key":"ref_4","first-page":"7","article-title":"Cancer statistics, 2019","volume":"69","author":"Siegel","year":"2019","journal-title":"CA Cancer J. Clin."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"719","DOI":"10.1038\/nrc3144","article-title":"Rethinking ovarian cancer: Recommendations for improving outcomes","volume":"11","author":"Vaughan","year":"2011","journal-title":"Nat. Rev. Cancer"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"260","DOI":"10.1016\/j.ygyno.2016.03.010","article-title":"Characteristics of 10-year survivors of high-grade serous ovarian carcinoma","volume":"141","author":"Dao","year":"2016","journal-title":"Gynecol. Oncol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2182","DOI":"10.1016\/S0140-6736(21)00731-5","article-title":"Ovarian cancer population screening and mortality after long-term follow-up in the UK Collaborative Trial of Ovarian Cancer Screening (UKCTOCS): A randomised controlled trial","volume":"397","author":"Menon","year":"2021","journal-title":"Lancet"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/j.ygyno.2018.08.016","article-title":"Histotype classification of ovarian carcinoma: A comparison of approaches","volume":"151","author":"Peres","year":"2018","journal-title":"Gynecol. Oncol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"4309","DOI":"10.1158\/1078-0432.CCR-18-3818","article-title":"Proteomics derived biomarker panel improves diagnostic precision to classify endometrioid and high-grade serous ovarian carcinoma","volume":"25","author":"Postovit","year":"2019","journal-title":"Clin. Cancer Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1093","DOI":"10.1038\/s41467-017-00962-1","article-title":"High grade serous ovarian carcinomas originate in the fallopian tube","volume":"8","author":"Papp","year":"2017","journal-title":"Nat. Commun."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"709049","DOI":"10.1155\/2012\/709049","article-title":"Early detection biomarkers for ovarian cancer","volume":"2012","author":"Sarojini","year":"2012","journal-title":"J. Oncol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1097\/IGC.0000000000000586","article-title":"Clinical Use of Cancer Biomarkers in Epithelial Ovarian Cancer: Updated Guidelines From the European Group on Tumor Markers","volume":"26","author":"Soletormos","year":"2016","journal-title":"Int. J. Gynecol. Cancer"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1065","DOI":"10.1158\/1078-0432.CCR-07-1569","article-title":"Diagnostic markers for early detection of ovarian cancer","volume":"14","author":"Visintin","year":"2008","journal-title":"Clin. Cancer Res."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"513","DOI":"10.2217\/17520363.1.4.513","article-title":"CA125 in ovarian cancer","volume":"1","author":"Scholler","year":"2007","journal-title":"Biomark. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"3628","DOI":"10.1158\/1078-0432.CCR-15-2750","article-title":"Early Detection of Ovarian Cancer using the Risk of Ovarian Cancer Algorithm with Frequent CA125 Testing in Women at Increased Familial Risk\u2014Combined Results from Two Screening Trials","volume":"23","author":"Skates","year":"2017","journal-title":"Clin. Cancer Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"430","DOI":"10.1159\/000442288","article-title":"CA125 and HE4: Measurement Tools for Ovarian Cancer","volume":"81","author":"Zhao","year":"2016","journal-title":"Gynecol. Obstet. Investig."},{"key":"ref_17","first-page":"239204","article-title":"Proteomics: Challenges, techniques and possibilities to overcome biological sample complexity","volume":"1","author":"Chandramouli","year":"2009","journal-title":"Hum. Genom. Proteom."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"M110-006353","DOI":"10.1074\/mcp.M110.006353","article-title":"A high-confidence human plasma proteome reference set with estimated concentrations in PeptideAtlas","volume":"10","author":"Farrah","year":"2011","journal-title":"Mol. Cell. Proteom."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1016\/j.ygyno.2016.03.036","article-title":"The biological significance and clinical applications of exosomes in ovarian cancer","volume":"142","author":"Dorayappan","year":"2016","journal-title":"Gynecol. Oncol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1038\/nature14581","article-title":"Glypican-1 identifies cancer exosomes and detects early pancreatic cancer","volume":"523","author":"Melo","year":"2015","journal-title":"Nature"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"eaau6977","DOI":"10.1126\/science.aau6977","article-title":"The biology, function, and biomedical applications of exosomes","volume":"367","author":"Kalluri","year":"2020","journal-title":"Science"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.ygyno.2008.05.020","article-title":"Exosomal microRNAs step into the biomarker arena","volume":"110","author":"Shih","year":"2008","journal-title":"Gynecol. Oncol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"631","DOI":"10.1038\/s41556-021-00693-y","article-title":"Quantitative proteomics identifies the core proteome of exosomes with syntenin-1 as the highest abundant protein and a putative universal biomarker","volume":"23","author":"Kugeratski","year":"2021","journal-title":"Nat. Cell Biol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1208","DOI":"10.1172\/JCI81135","article-title":"The biology and function of exosomes in cancer","volume":"126","author":"Kalluri","year":"2016","journal-title":"J. Clin. Investig."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"382","DOI":"10.1038\/s41586-018-0392-8","article-title":"Exosomal PD-L1 contributes to immunosuppression and is associated with anti-PD-1 response","volume":"560","author":"Chen","year":"2018","journal-title":"Nature"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"498","DOI":"10.1038\/nature22341","article-title":"Exosomes facilitate therapeutic targeting of oncogenic KRAS in pancreatic cancer","volume":"546","author":"Kamerkar","year":"2017","journal-title":"Nature"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"3503","DOI":"10.1158\/0008-5472.CAN-18-3538","article-title":"A Microfluidic Chip Enables Isolation of Exosomes and Establishment of Their Protein Profiles and Associated Signaling Pathways in Ovarian Cancer","volume":"79","author":"Dorayappan","year":"2019","journal-title":"Cancer Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"3144","DOI":"10.1039\/C8LC00834E","article-title":"Microfluidic affinity separation chip for selective capture and release of label-free ovarian cancer exosomes","volume":"18","author":"Hisey","year":"2018","journal-title":"Lab. A Chip"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2609","DOI":"10.1038\/s41592-025-02914-w","article-title":"Light-induced extracellular vesicle and particle adsorption","volume":"22","author":"Hisey","year":"2025","journal-title":"Nat. Methods"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"3806","DOI":"10.1038\/s41388-018-0189-0","article-title":"Hypoxia-induced exosomes contribute to a more aggressive and chemoresistant ovarian cancer phenotype: A novel mechanism linking STAT3\/Rab proteins","volume":"37","author":"Dorayappan","year":"2018","journal-title":"Oncogene"},{"key":"ref_31","first-page":"4399","article-title":"Nanouric acid or nanocalcium phosphate as central nidus to induce calcium oxalate stone formation: A high-resolution transmission electron microscopy study on urinary nanocrystallites","volume":"9","author":"Gao","year":"2014","journal-title":"Int. J. Nanomed."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1093\/biostatistics\/1.2.123","article-title":"Combining diagnostic test results to increase accuracy","volume":"1","author":"Pepe","year":"2000","journal-title":"Biostatistics"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"242","DOI":"10.1177\/09622802221137742","article-title":"A distribution-free smoothed combination method to improve discrimination accuracy in multi-category classification","volume":"32","author":"Maiti","year":"2023","journal-title":"Stat. Methods Med. Res."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Das, P., De, D., Maiti, R., Kamal, M., Hutcheson, K.A., Fuller, C.D., Chakraborty, B., and Peterson, C.B. (2022). Estimating the optimal linear combination of predictors using spherically constrained optimization. BMC Bioinform., 23.","DOI":"10.1186\/s12859-022-04953-y"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1002\/1097-0142(1950)3:1<32::AID-CNCR2820030106>3.0.CO;2-3","article-title":"Index for rating diagnostic tests","volume":"3","author":"Youden","year":"1950","journal-title":"Cancer"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"112280","DOI":"10.1016\/j.isci.2025.112280","article-title":"Catalyzing early ovarian cancer detection: Platelet RNA-based precision screening","volume":"28","author":"Ahn","year":"2025","journal-title":"iScience"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1016\/j.ygyno.2025.08.012","article-title":"Prevention, early detection & interception: 15th biennial ovarian cancer research symposium","volume":"201","author":"Sasamoto","year":"2025","journal-title":"Gynecol. Oncol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1516","DOI":"10.1158\/2767-9764.CRC-25-0140","article-title":"Utilizing Serum-Derived Lipidomics with Protein Biomarkers and Machine Learning for Early Detection of Ovarian Cancer in the Symptomatic Population","volume":"5","author":"Giles","year":"2025","journal-title":"Cancer Res. Commun."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2062","DOI":"10.1200\/JCO.2014.59.4945","article-title":"Risk Algorithm Using Serial Biomarker Measurements Doubles the Number of Screen-Detected Cancers Compared With a Single-Threshold Rule in the United Kingdom Collaborative Trial of Ovarian Cancer Screening","volume":"33","author":"Menon","year":"2015","journal-title":"J. Clin. Oncol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1180","DOI":"10.1158\/2159-8290.CD-24-1366","article-title":"Multimodal Spatial Profiling Reveals Immune Suppression and Microenvironment Remodeling in Fallopian Tube Precursors to High-Grade Serous Ovarian Carcinoma","volume":"15","author":"Kader","year":"2025","journal-title":"Cancer Discov."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1158\/2159-8290.CD-24-0393","article-title":"Early Detection of Ovarian Cancer Using Cell-Free DNA Fragmentomes and Protein Biomarkers","volume":"15","author":"Medina","year":"2025","journal-title":"Cancer Discov."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1739","DOI":"10.1158\/0008-5472.CAN-17-1671","article-title":"STAT3\/PIAS3 Levels Serve as \u201cEarly Signature\u201d Genes in the Development of High-Grade Serous Carcinoma from the Fallopian Tube","volume":"78","author":"Saini","year":"2018","journal-title":"Cancer Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1141","DOI":"10.1158\/0008-5472.CAN-13-2247","article-title":"Cyclin E1 deregulation occurs early in secretory cell transformation to promote formation of fallopian tube-derived high-grade serous ovarian cancers","volume":"74","author":"Karst","year":"2014","journal-title":"Cancer Res."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"609","DOI":"10.1038\/nature10166","article-title":"Integrated genomic analyses of ovarian carcinoma","volume":"474","year":"2011","journal-title":"Nature"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"9883","DOI":"10.2147\/CMAR.S269215","article-title":"Prognostic Significance of Pregnancy Zone Protein and Its Correlation with Immune Infiltrates in Hepatocellular Carcinoma","volume":"12","author":"Su","year":"2020","journal-title":"Cancer Manag. Res."},{"key":"ref_46","first-page":"161","article-title":"Human complement factor H is a novel diagnostic marker for lung adenocarcinoma","volume":"39","author":"Cui","year":"2011","journal-title":"Int. J. Oncol."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/15\/8\/706\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T10:53:01Z","timestamp":1777459981000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/15\/8\/706"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,16]]},"references-count":46,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2026,4]]}},"alternative-id":["cells15080706"],"URL":"https:\/\/doi.org\/10.3390\/cells15080706","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,16]]}}}