{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,5,1]],"date-time":"2025-05-01T23:10:10Z","timestamp":1746141010534,"version":"3.40.4"},"reference-count":37,"publisher":"Georg Thieme Verlag KG","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Thromb Haemost"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Protein C (PC) activation on endothelial cells is a critical antithrombotic mechanism. Hereditary PC deficiency (PCD), which is caused by mutations in the PROC gene, can predispose affected individuals to thrombophilia. Previous studies investigated activated protein C (APC) generation in PCD patients without including endothelial cells, which are essential for physiological PC activation. This study aimed to assess APC generation in PCD patients using a novel endothelial cell-based assay.<\/jats:p><jats:p>Plasma samples from 21 patients with 19 heterozygous PROC mutations (median PC level 58%) and 24 healthy controls were analyzed. Endothelium-dependent APC generation was initiated by overlaying plasma on human umbilical vein endothelial cells (HUVECs) and adding tissue factor (1 pmol\/L). APC levels were quantified using an oligonucleotide-based enzyme capture assay. The area under the curve (AUC) was calculated to monitor cumulative APC formation over time. A calibration curve generated from wild-type PC in PC-deficient plasma established reference ranges.<\/jats:p><jats:p>Mean peak levels of APC were significantly lower in PCD patients than in healthy controls (0.75 vs. 1.83\u2009nmol\/L, p\u2009=\u20092\u2009\u00d7\u200910\u221210). The AUC APC was below the reference range in 8 of 21 (38%) patient samples, indicating disproportionately severe impairment in APC generation. The observed variability in APC generation suggests that endothelial contributions may identify functional differences undetected by standard PC activity or antigen assays.<\/jats:p><jats:p>This study introduces a novel endothelial cell-based APC generation assay, demonstrating the functional consequences of PROC mutations and providing insights into the regulation of APC generation, with potential applications in thrombosis risk assessment and personalized therapy.<\/jats:p>","DOI":"10.1055\/a-2579-6390","type":"journal-article","created":{"date-parts":[[2025,4,10]],"date-time":"2025-04-10T23:10:15Z","timestamp":1744326615000},"source":"Crossref","is-referenced-by-count":0,"title":["Endothelium-Dependent Protein C Activation in Hereditary Protein C Deficiency"],"prefix":"10.1055","author":[{"given":"Nadine","family":"Schwarz","sequence":"additional","affiliation":[{"name":"Institute of Experimental Hematology and Transfusion Medicine, University Hospital Bonn, Bonn, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hannah L.","family":"McRae","sequence":"additional","affiliation":[{"name":"Institute of Experimental Hematology and Transfusion Medicine, University Hospital Bonn, Bonn, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sara","family":"Reda","sequence":"additional","affiliation":[{"name":"Institute of Experimental Hematology and Transfusion Medicine, University Hospital Bonn, Bonn, Germany"},{"name":"Department of Transfusion Medicine, University Hospital Cologne, Cologne, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4796-3424","authenticated-orcid":false,"given":"Behnaz","family":"Pezeshkpoor","sequence":"additional","affiliation":[{"name":"Institute of Experimental Hematology and Transfusion Medicine, University Hospital Bonn, Bonn, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Johannes","family":"Oldenburg","sequence":"additional","affiliation":[{"name":"Institute of Experimental Hematology and Transfusion Medicine, University Hospital Bonn, Bonn, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jens","family":"M\u00fcller","sequence":"additional","affiliation":[{"name":"Institute of Experimental Hematology and Transfusion Medicine, University Hospital Bonn, Bonn, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bernd","family":"P\u00f6tzsch","sequence":"additional","affiliation":[{"name":"Institute of Experimental Hematology and Transfusion Medicine, University Hospital Bonn, Bonn, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Heiko","family":"R\u00fchl","sequence":"additional","affiliation":[{"name":"Institute of Experimental Hematology and Transfusion Medicine, University Hospital Bonn, Bonn, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"194","published-online":{"date-parts":[[2025,5,1]]},"reference":[{"issue":"15","key":"ref1","doi-asserted-by":"crossref","first-page":"3310","DOI":"10.1016\/j.febslet.2005.03.001","article-title":"The anticoagulant protein C pathway","volume":"579","author":"B Dahlb\u00e4ck","year":"2005","journal-title":"FEBS Lett"},{"issue":"S 01","key":"ref2","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1111\/j.1538-7836.2007.02491.x","article-title":"Activated protein C","volume":"5","author":"J H Griffin","year":"2007","journal-title":"J Thromb Haemost"},{"issue":"07","key":"ref3","doi-asserted-by":"crossref","first-page":"1311","DOI":"10.1161\/01.ATV.0000168421.13467.82","article-title":"Regulation of blood coagulation by the protein C anticoagulant pathway: novel insights into structure-function relationships and molecular recognition","volume":"25","author":"B Dahlb\u00e4ck","year":"2005","journal-title":"Arterioscler Thromb Vasc Biol"},{"issue":"19","key":"ref4","doi-asserted-by":"crossref","first-page":"2059","DOI":"10.2174\/092986710791233706","article-title":"Regulation of the protein C anticoagulant and antiinflammatory pathways","volume":"17","author":"A R Rezaie","year":"2010","journal-title":"Curr Med Chem"},{"issue":"03","key":"ref5","first-page":"217","article-title":"Resistance to activated protein C as risk factor for thrombosis: molecular mechanisms, laboratory investigation, and clinical management","volume":"34","author":"B Dahlb\u00e4ck","year":"1997","journal-title":"Semin Hematol"},{"issue":"04","key":"ref6","doi-asserted-by":"crossref","first-page":"579","DOI":"10.1046\/j.1365-2141.1996.d01-1954.x","article-title":"The population genetics of factor V Leiden (Arg506Gln)","volume":"95","author":"D C Rees","year":"1996","journal-title":"Br J Haematol"},{"issue":"06","key":"ref7","doi-asserted-by":"crossref","first-page":"1214","DOI":"10.1111\/j.1365-2516.2008.01838.x","article-title":"Protein C deficiency","volume":"14","author":"N A Goldenberg","year":"2008","journal-title":"Haemophilia"},{"issue":"01","key":"ref8","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/0049-3848(94)00138-4","article-title":"The protein C anticoagulant system: inherited defects as basis for venous thrombosis","volume":"77","author":"B Dahlb\u00e4ck","year":"1995","journal-title":"Thromb Res"},{"issue":"10","key":"ref9","doi-asserted-by":"crossref","first-page":"1281","DOI":"10.5858\/arpa.2017-0403-RS","article-title":"Deficiency","volume":"143","author":"P Dinarvand","year":"2019","journal-title":"Arch Pathol Lab Med"},{"issue":"03","key":"ref10","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1111\/j.1365-2141.2008.07339.x","article-title":"Does thrombophilia testing help in the clinical management of patients?","volume":"143","author":"S Middeldorp","year":"2008","journal-title":"Br J Haematol"},{"issue":"02","key":"ref11","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1111\/jth.14667","article-title":"Recommendations for clinical laboratory testing for protein C deficiency, for the subcommittee on plasma coagulation inhibitors of the ISTH","volume":"18","author":"P C Cooper","year":"2020","journal-title":"J Thromb Haemost"},{"issue":"01","key":"ref12","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1111\/ijlh.12004","article-title":"Guidelines on the laboratory aspects of assays used in haemostasis and thrombosis","volume":"35","author":"I Mackie","year":"2013","journal-title":"Int J Lab Hematol"},{"issue":"06","key":"ref13","doi-asserted-by":"crossref","first-page":"440","DOI":"10.1002\/ajh.21679","article-title":"Laboratory tests for protein C deficiency","volume":"85","author":"B Khor","year":"2010","journal-title":"Am J Hematol"},{"issue":"04","key":"ref14","doi-asserted-by":"crossref","first-page":"748","DOI":"10.1046\/j.1538-7836.2003.00141.x","article-title":"The long-term within- and between-laboratory variability for assay of antithrombin, and proteins C and S: results derived from the external quality assessment program for thrombophilia screening of the ECAT Foundation","volume":"1","author":"P Meijer","year":"2003","journal-title":"J Thromb Haemost"},{"issue":"06","key":"ref15","doi-asserted-by":"crossref","first-page":"909","DOI":"10.1309\/AJCP8MWU4QSTCLPU","article-title":"Protein C assay performance: an analysis of North American specialized coagulation laboratory association proficiency testing results","volume":"137","author":"J M Baron","year":"2012","journal-title":"Am J Clin Pathol"},{"issue":"06","key":"ref16","doi-asserted-by":"crossref","first-page":"2033","DOI":"10.1172\/JCI111626","article-title":"Detection of protein C activation in humans","volume":"74","author":"K A Bauer","year":"1984","journal-title":"J Clin Invest"},{"issue":"05","key":"ref17","doi-asserted-by":"crossref","first-page":"1418","DOI":"10.1182\/blood.V71.5.1418.1418","article-title":"Hemostatic enzyme generation in the blood of patients with hereditary protein C deficiency","volume":"71","author":"K A Bauer","year":"1988","journal-title":"Blood"},{"issue":"09","key":"ref18","doi-asserted-by":"crossref","first-page":"2340","DOI":"10.1182\/blood.V79.9.2340.2340","article-title":"Direct detection of activated protein C in blood from human subjects","volume":"79","author":"A Gruber","year":"1992","journal-title":"Blood"},{"issue":"04","key":"ref19","doi-asserted-by":"crossref","first-page":"1159","DOI":"10.1182\/blood.V82.4.1159.1159","article-title":"Normalization of markers of coagulation activation with a purified protein C concentrate in adults with homozygous protein C deficiency","volume":"82","author":"J Conard","year":"1993","journal-title":"Blood"},{"issue":"01","key":"ref20","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1161\/ATVBAHA.122.318433","article-title":"Ex vivo modeling of the PC (protein C) pathway using endothelial cells and plasma: a personalized approach","volume":"43","author":"N Schwarz","year":"2023","journal-title":"Arterioscler Thromb Vasc Biol"},{"issue":"27","key":"ref21","doi-asserted-by":"crossref","first-page":"6075","DOI":"10.1002\/anie.201007032","article-title":"Profiling of active thrombin in human blood by supramolecular complexes","volume":"50","author":"J M\u00fcller","year":"2011","journal-title":"Angew Chem Int Ed Engl"},{"issue":"03","key":"ref22","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1111\/j.1538-7836.2012.04623.x","article-title":"Monitoring of plasma levels of activated protein C using a clinically applicable oligonucleotide-based enzyme capture assay","volume":"10","author":"J M\u00fcller","year":"2012","journal-title":"J Thromb Haemost"},{"issue":"19","key":"ref23","doi-asserted-by":"crossref","first-page":"10212","DOI":"10.1073\/pnas.93.19.10212","article-title":"The endothelial cell protein C receptor augments protein C activation by the thrombin-thrombomodulin complex","volume":"93","author":"D J Stearns-Kurosawa","year":"1996","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"07","key":"ref24","doi-asserted-by":"crossref","first-page":"1029","DOI":"10.1084\/jem.187.7.1029","article-title":"Activation mechanism of anticoagulant protein C in large blood vessels involving the endothelial cell protein C receptor","volume":"187","author":"K Fukudome","year":"1998","journal-title":"J Exp Med"},{"issue":"04","key":"ref25","doi-asserted-by":"crossref","first-page":"614","DOI":"10.1160\/TH12-10-0760","article-title":"Expression and functional characterisation of natural R147W and K150del variants of protein C in the Chinese population","volume":"109","author":"Q Ding","year":"2013","journal-title":"Thromb Haemost"},{"issue":"15","key":"ref26","doi-asserted-by":"crossref","first-page":"2428","DOI":"10.1182\/blood-2014-12-617274","article-title":"Protein C Thr315Ala variant results in gain of function but manifests as type II deficiency in diagnostic assays","volume":"125","author":"Q Ding","year":"2015","journal-title":"Blood"},{"issue":"07","key":"ref27","doi-asserted-by":"crossref","first-page":"1358","DOI":"10.1160\/TH17-01-0043","article-title":"Gly74Ser mutation in protein C causes thrombosis due to a defect in protein S-dependent anticoagulant function","volume":"117","author":"C Chen","year":"2017","journal-title":"Thromb Haemost"},{"issue":"05","key":"ref28","doi-asserted-by":"crossref","first-page":"1141","DOI":"10.1111\/jth.14777","article-title":"Gly197Arg mutation in protein C causes recurrent thrombosis in a heterozygous carrier","volume":"18","author":"Y Lu","year":"2020","journal-title":"J Thromb Haemost"},{"issue":"06","key":"ref29","doi-asserted-by":"crossref","first-page":"129892","DOI":"10.1016\/j.bbagen.2021.129892","article-title":"Role of Gly197 in the structure and function of protein C","volume":"1865","author":"Y Lu","year":"2021","journal-title":"Biochim Biophys Acta, Gen Subj"},{"issue":"03","key":"ref30","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1182\/blood.V63.3.671.671","article-title":"A functional assay of protein C in human plasma","volume":"63","author":"N Sala","year":"1984","journal-title":"Blood"},{"issue":"02","key":"ref31","doi-asserted-by":"crossref","first-page":"416","DOI":"10.1172\/JCI112319","article-title":"Relationship between protein C antigen and anticoagulant activity during oral anticoagulation and in selected disease states","volume":"77","author":"S Vigano D'Angelo","year":"1986","journal-title":"J Clin Invest"},{"key":"ref32","doi-asserted-by":"crossref","first-page":"755281","DOI":"10.3389\/fcvm.2021.755281","article-title":"PC deficiency testing: thrombin-thrombomodulin as PC activator and aptamer-based enzyme capturing increase diagnostic accuracy","volume":"8","author":"S Reda","year":"2021","journal-title":"Front Cardiovasc Med"},{"key":"ref33","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/j.febslet.2004.06.060","article-title":"Functional characterization of recombinant batroxobin, a snake venom thrombin-like enzyme, expressed from Pichia pastoris","volume":"571","author":"W K You","year":"2004","journal-title":"FEBS Lett"},{"key":"ref34","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1111\/j.1538-7836.2007.02516.x","article-title":"Serpins in thrombosis, hemostasis and fibrinolysis","volume":"5","author":"J C Rau","year":"2007","journal-title":"J Thromb 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