{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,1]],"date-time":"2026-05-01T00:53:45Z","timestamp":1777596825409,"version":"3.51.4"},"reference-count":23,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2021,1,14]],"date-time":"2021-01-14T00:00:00Z","timestamp":1610582400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100015739","name":"Bayer Vital GmbH","doi-asserted-by":"publisher","award":["121213"],"award-info":[{"award-number":["121213"]}],"id":[{"id":"10.13039\/100015739","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Annelise-Asmussen Foundation","award":["20139592"],"award-info":[{"award-number":["20139592"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The development of malignant effusions such as ascites reflects a massive progression of a malignant disease. In patients with ovarian carcinoma, a high amount of ascites (&gt;500 mL) is an independent negative prognostic marker. The composition and constituents of ascites reflect the inflammatory environment of the underlying tumor. Increased cellular resistance of ascites-derived tumor cells and the development of venous thromboembolic events (VTE) are major risks for these patients, especially in patients with advanced ovarian carcinoma. In this study, we discuss the release of tissue factor-bearing extracellular vesicles (TF+ EVs) from tumor cells into the environment (ascites fluid) and their systemic spreading as a possible causal explanation of the pathologic coagulation status in these patients. We obtained ascites from patients with advanced ovarian carcinoma, collected during surgery or therapeutic paracentesis (n = 20). Larger ectosome-like EVs were isolated using sequential centrifugation, quantified by high-resolution flow cytometry and analyzed using nanoparticle tracking analysis. Furthermore, the pro-coagulant properties (TF activity) of EVs were determined. Compared to published TF activities of EVs from healthy persons, TF activities of EVs derived from ascites of patients with ovarian cancer were very high, with a median of 80 pg\/mL. The rate of VTE, as reported in the patient files, was high as well (35%, 7 out of 20). Furthermore, all but one patient with VTE had EV concentrations above the median within their ascetic fluid (p &lt; 0.02). Since VTE continues to be a frequent cause of death in cancer patients, prophylactic antithrombotic treatment might be worth considering in these patients. However, given the risk of bleeding, more clinical data are warranted. Although the study is too small to enable reaching a conclusion on direct clinical implementation, it can well serve as a proof of principle and a rationale to initiate a prospective clinical study with different patient subgroups. We also show ex vivo that these larger ectosome-like EVs induce intracellular ERK phosphorylation and tumor cell migration, which is not directly related to their pro-coagulative potency, but might help to understand why cancer patients with thromboembolic events have a poorer prognosis.<\/jats:p>","DOI":"10.3390\/ijms22020790","type":"journal-article","created":{"date-parts":[[2021,1,14]],"date-time":"2021-01-14T11:39:14Z","timestamp":1610624354000},"page":"790","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Biologically Active Tissue Factor-Bearing Larger Ectosome-Like Extracellular Vesicles in Malignant Effusions from Ovarian Cancer Patients: Correlation with Incidence of Thrombosis"],"prefix":"10.3390","volume":"22","author":[{"given":"Corinna","family":"Steidel","sequence":"first","affiliation":[{"name":"Clinic for Hematology and Oncology, University Hospital and Medical School (UKSH), University of Luebeck, Ratzeburger Allee 160, 23538 Luebeck, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fanny","family":"Ender","sequence":"additional","affiliation":[{"name":"Clinic for Hematology and Oncology, University Hospital and Medical School (UKSH), University of Luebeck, Ratzeburger Allee 160, 23538 Luebeck, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Achim","family":"Rody","sequence":"additional","affiliation":[{"name":"Department of Gynecology and Obstetrics, University Hospital and Medical School (UKSH), Ratzeburger Allee 160, 23538 Luebeck, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nikolas","family":"von Bubnoff","sequence":"additional","affiliation":[{"name":"Clinic for Hematology and Oncology, University Hospital and Medical School (UKSH), University of Luebeck, Ratzeburger Allee 160, 23538 Luebeck, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6409-3822","authenticated-orcid":false,"given":"Frank","family":"Gieseler","sequence":"additional","affiliation":[{"name":"Clinic for Hematology and Oncology, University Hospital and Medical School (UKSH), University of Luebeck, Ratzeburger Allee 160, 23538 Luebeck, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,1,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2528","DOI":"10.1182\/blood-2018-99-110271","article-title":"Case Fatality Rate of Recurrent Venous Thromboembolism and Major Bleeding Events Among Patients Treated for Cancer-Associated Venous Thromboembolism: A Systematic Review","volume":"132","author":"Abdulla","year":"2018","journal-title":"Blood"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Weeks, K.S., Herbach, E., McDonald, M., Charlton, M., and Schweizer, M.L. (2020). Meta-Analysis of VTE Risk: Ovarian Cancer Patients by Stage, Histology, Cytoreduction, and Ascites at Diagnosis. Obstet. Gynecol. Int.","DOI":"10.1155\/2020\/2374716"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1050","DOI":"10.1002\/cbin.10645","article-title":"Extracellular vesicles from malignant effusions induce tumor cell migration: Inhibitory effect of LMWH tinzaparin","volume":"40","author":"Gamperl","year":"2016","journal-title":"Cell Biol. Int."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"167","DOI":"10.5306\/wjco.v9.i8.167","article-title":"Malignant peritoneal effusion acting as a tumor environment in ovarian cancer progression: Impact and significance","volume":"9","year":"2018","journal-title":"World J. Clin. Oncol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1015","DOI":"10.1161\/01.ATV.0000130465.23430.74","article-title":"Role of tissue factor in hemostasis, thrombosis, and vascular development","volume":"24","author":"Mackman","year":"2004","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"546","DOI":"10.1097\/IGC.0b013e318241d9b9","article-title":"Microparticles from ovarian carcinomas are shed into ascites and promote cell migration","volume":"22","author":"Press","year":"2012","journal-title":"Int. J. Gynecol. Cancer"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1160\/th15-03-0213","article-title":"High-level secretion of tissue factor-rich extracellular vesicles from ovarian cancer cells mediated by filamin-A and protease-activated receptors","volume":"115","author":"Koizume","year":"2016","journal-title":"Thromb. Haemost."},{"key":"ref_8","first-page":"3439","article-title":"Membrane vesicles in ovarian cancer fluids: A new potential marker","volume":"19","author":"Ginestra","year":"1999","journal-title":"Anticancer Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1535750","DOI":"10.1080\/20013078.2018.1535750","article-title":"Minimal information for studies of extracellular vesicles 2018 (MISEV2018): A position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines","volume":"7","author":"Witwer","year":"2018","journal-title":"J. Extracell. Vesicles"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1038\/nrc3432","article-title":"Meeting the challenge of ascites in ovarian cancer: New avenues for therapy and research","volume":"13","author":"Kipps","year":"2013","journal-title":"Nat. Rev. Cancer"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"563","DOI":"10.1016\/j.ygyno.2007.08.064","article-title":"Malignant ascites-derived exosomes of ovarian carcinoma patients contain CD24 and EpCAM","volume":"107","author":"Runz","year":"2007","journal-title":"Gynecol. Oncol."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Ender, F., Zamzow, P., von Bubnoff, N., and Gieseler, F. (2020). Detection and Quantification of Extracellular Vesicles via FACS: Membrane Labeling Matters!. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21010291"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/1478-811X-11-86","article-title":"Proteinase-activated receptors (PARs)\u2013focus on receptor-receptor-interactions and their physiological and pathophysiological impact","volume":"11","author":"Gieseler","year":"2013","journal-title":"Cell Commun. Signal."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1002\/rth2.12165","article-title":"Measurement of tissue factor activity in extracellular vesicles from human plasma samples","volume":"3","author":"Hisada","year":"2019","journal-title":"Res. Pract. Thromb. Haemost."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1006\/gyno.1996.0106","article-title":"The prognostic implication of ascites in advanced-stage ovarian cancer","volume":"61","author":"Puls","year":"1996","journal-title":"Gynecol. Oncol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"S143","DOI":"10.1016\/j.thromres.2018.01.023","article-title":"Tissue factor as a mediator of coagulation and signaling in cancer and chronic inflammation","volume":"164","author":"Graf","year":"2018","journal-title":"Thromb. Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1506","DOI":"10.1093\/annonc\/mdm190","article-title":"Role of protease activated receptor-2 in tumor advancement of ovarian cancers","volume":"18","author":"Jahan","year":"2007","journal-title":"Ann. Oncol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1024","DOI":"10.1007\/s00018-005-5570-9","article-title":"Cancer and blood coagulation","volume":"63","author":"Boccaccio","year":"2006","journal-title":"Cell. Mol. Life Sci. CMLS"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Lacroix, R., Judicone, C., Mooberry, M., Boucekine, M., Key, N.S., and Dignat-George, F. (2013). Standardization of pre-analytical variables in plasma microparticle determination: Results of the International Society on Thrombosis and Haemostasis SSC Collaborative workshop. J. Thromb. Haemost. JTH.","DOI":"10.1111\/jth.12207"},{"key":"ref_20","first-page":"1875","article-title":"ARF6-regulated shedding of tumor cell-derived plasma membrane microvesicles","volume":"10","author":"Clancy","year":"2009","journal-title":"J. Cell Sci."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1111\/jth.12767","article-title":"A simple flow cytometry method improves the detection of phosphatidylserine-exposing extracellular vesicles","volume":"13","author":"Arraud","year":"2015","journal-title":"J. Thromb. Haemost."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"591","DOI":"10.1002\/ijc.2910250507","article-title":"Human cell line (COLO 357) of metastatic pancreatic adenocarcinoma","volume":"25","author":"Morgan","year":"1980","journal-title":"Int. J. Cancer"},{"key":"ref_23","first-page":"5978","article-title":"\u201cCOLO 357,\u201d a human pancreatic adenosquamous carcinoma: Growth in artificial capillary culture and in nude mice","volume":"43","author":"Meitner","year":"1983","journal-title":"Cancer Res."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/2\/790\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:11:10Z","timestamp":1760159470000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/2\/790"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,1,14]]},"references-count":23,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2021,1]]}},"alternative-id":["ijms22020790"],"URL":"https:\/\/doi.org\/10.3390\/ijms22020790","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,1,14]]}}}