{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,3]],"date-time":"2026-05-03T11:01:07Z","timestamp":1777806067852,"version":"3.51.4"},"reference-count":28,"publisher":"MDPI AG","issue":"22","license":[{"start":{"date-parts":[[2022,11,8]],"date-time":"2022-11-08T00:00:00Z","timestamp":1667865600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005416","name":"Research Council of Norway","doi-asserted-by":"publisher","award":["251239"],"award-info":[{"award-number":["251239"]}],"id":[{"id":"10.13039\/501100005416","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Factor-VII-activating protease (FSAP) is involved in the regulation of hemostasis and inflammation. Extracellular histones play a role in inflammation and the conversion of latent pro-FSAP into active FSAP. FSAP has been shown to regulate endothelial permeability, but the mechanisms are not clear. Here, we have investigated the effects of FSAP on endothelial permeability in vitro. A mixture of histones from calf thymus stimulated permeability, and the wild-type (WT) serine protease domain (SPD) of FSAP blocked this effect. WT\u2013SPD\u2013FSAP did not influence permeability on its own, nor that stimulated by thrombin or vascular endothelial growth factor (VEGF)-A165. Histones induced a large-scale rearrangement of the junction proteins VE-cadherin and zona occludens-1 from a clear junctional distribution to a diffuse pattern. The presence of WT\u2013SPD\u2013FSAP inhibited these changes. Permeability changes by histones were blocked by both TLR-2 and TLR4 blocking antibodies. Histones upregulated the expression of TLR-2, but not TLR-4, in HUVEC cells, and WT\u2013SPD\u2013FSAP abolished the upregulation of TLR-2 expression. An inactive variant, Marburg I (MI)\u2013SPD\u2013FSAP, did not have any of these effects. The inhibition of histone-mediated permeability may be an important function of FSAP with relevance to sepsis, trauma, and stroke and the need to be investigated further in in vivo experiments.<\/jats:p>","DOI":"10.3390\/ijms232213706","type":"journal-article","created":{"date-parts":[[2022,11,8]],"date-time":"2022-11-08T08:52:56Z","timestamp":1667897576000},"page":"13706","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["FSAP Protects against Histone-Mediated Increase in Endothelial Permeability In Vitro"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7335-5799","authenticated-orcid":false,"given":"Xue Yan","family":"Cui","sequence":"first","affiliation":[{"name":"Department of Haematology, Oslo University Hospital, 0424 Oslo, Norway"},{"name":"Research Institute of Internal Medicine, Oslo University Hospital, 0424 Oslo, Norway"},{"name":"Institute of Clinical Medicine, University of Oslo, 0315 Oslo, Norway"},{"name":"Department of Haematology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3341-9847","authenticated-orcid":false,"given":"Benedicte","family":"Stavik","sequence":"additional","affiliation":[{"name":"Department of Haematology, Oslo University Hospital, 0424 Oslo, Norway"},{"name":"Research Institute of Internal Medicine, Oslo University Hospital, 0424 Oslo, Norway"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2804-9522","authenticated-orcid":false,"given":"Bernd","family":"Thiede","sequence":"additional","affiliation":[{"name":"Department of Biosciences, University of Oslo, 0315 Oslo, Norway"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5556-8099","authenticated-orcid":false,"given":"Per Morten","family":"Sandset","sequence":"additional","affiliation":[{"name":"Department of Haematology, Oslo University Hospital, 0424 Oslo, Norway"},{"name":"Research Institute of Internal Medicine, Oslo University Hospital, 0424 Oslo, Norway"},{"name":"Institute of Clinical Medicine, University of Oslo, 0315 Oslo, Norway"},{"name":"Department of Haematology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0782-9957","authenticated-orcid":false,"given":"Sandip M.","family":"Kanse","sequence":"additional","affiliation":[{"name":"Institute of Basic Medical Sciences, University of Oslo, 0315 Oslo, Norway"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,11,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"314","DOI":"10.1016\/j.molmed.2020.11.006","article-title":"Permeability of the Endothelial Barrier: Identifying and Reconciling Controversies","volume":"27","author":"Dejana","year":"2021","journal-title":"Trends. Mol. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"562","DOI":"10.1002\/(SICI)1097-4652(199606)167:3<562::AID-JCP20>3.0.CO;2-4","article-title":"Mechanisms of alpha-thrombin, histamine, and bradykinin induced endothelial permeability","volume":"167","author":"Ehringer","year":"1996","journal-title":"J. Cell Physiol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"563","DOI":"10.1210\/me.2004-0496","article-title":"Epigenetic regulation by histone methylation and histone variants","volume":"19","author":"Cheung","year":"2005","journal-title":"Mol. Endocrinol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"507","DOI":"10.1007\/s11010-021-04300-4","article-title":"Extra-nuclear histones: Origin, significance and perspectives","volume":"477","author":"Singh","year":"2022","journal-title":"Mol. Cell Biochem."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1164\/rccm.201206-1037OC","article-title":"Circulating histones are mediators of trauma-associated lung injury","volume":"187","author":"Abrams","year":"2013","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e2518","DOI":"10.1038\/cddis.2016.410","article-title":"Extracellular histones, cell-free DNA, or nucleosomes: Differences in immunostimulation","volume":"7","author":"Marsman","year":"2016","journal-title":"Cell Death Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1375","DOI":"10.1681\/ASN.2011111077","article-title":"Histones from dying renal cells aggravate kidney injury via TLR2 and TLR4","volume":"23","author":"Allam","year":"2012","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Milone, G., Bellofiore, C., Leotta, S., Milone, G.A., Cupri, A., Duminuco, A., Garibaldi, B., and Palumbo, G. (2022). Endothelial Dysfunction after Hematopoietic Stem Cell Transplantation: A Review Based on Physiopathology. J. Clin. Med., 11.","DOI":"10.3390\/jcm11030623"},{"key":"ref_9","first-page":"424759","article-title":"Factor VII Activating Protease Polymorphism (G534E) Is Associated with Increased Risk for Stroke and Mortality","volume":"2011","author":"Trompet","year":"2011","journal-title":"Stroke Res. Treat."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"667","DOI":"10.1161\/01.CIR.0000055189.18831.B1","article-title":"Marburg I polymorphism of factor VII--activating protease: A prominent risk predictor of carotid stenosis","volume":"107","author":"Willeit","year":"2003","journal-title":"Circulation"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1119","DOI":"10.1515\/BC.2002.121","article-title":"Factor-VII-activating protease (FSAP): A novel protease in hemostasis","volume":"383","author":"Romisch","year":"2002","journal-title":"Biol. Chem."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"483","DOI":"10.1016\/j.bbrc.2011.05.030","article-title":"Extracellular histone induces plasma hyaluronan-binding protein (factor-VII-activating protease) activation in vivo","volume":"409","author":"Yamamichi","year":"2011","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2491","DOI":"10.1182\/bloodadvances.2017010959","article-title":"DNA and factor VII-activating protease protect against the cytotoxicity of histones","volume":"1","author":"Marsman","year":"2017","journal-title":"Blood Adv."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1750","DOI":"10.1160\/TH17-02-0081","article-title":"Analysis of the substrate specificity of Factor-VII-activating protease (FSAP) and design of specific and sensitive peptide substrates","volume":"117","author":"Kara","year":"2017","journal-title":"Thromb. Haemost."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1007\/s40291-020-00460-z","article-title":"Use of DAMPs and SAMPs as Therapeutic Targets or Therapeutics: A Note of Caution","volume":"24","author":"Land","year":"2020","journal-title":"Mol. Diagn. Ther."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"L223","DOI":"10.1152\/ajplung.00072.2022","article-title":"Circulating extracellular histones exacerbate acute lung injury by augmenting pulmonary endothelial dysfunction via TLR4-dependent mechanism","volume":"323","author":"Kim","year":"2022","journal-title":"Am. J. Physiol. Lung Cell Mol. Physiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1016\/j.thromres.2017.11.012","article-title":"Interaction of factor-VII-activating protease (FSAP) with neutrophil extracellular traps (NETs)","volume":"161","author":"Grasso","year":"2018","journal-title":"Thromb. Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1079","DOI":"10.1096\/fj.201801986RR","article-title":"Protease activated receptors (PAR)-1 and -2 mediate cellular effects of factor-VII-activating protease (FSAP)","volume":"34","author":"Byskov","year":"2020","journal-title":"FASEB J."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"217","DOI":"10.1007\/s00424-021-02639-7","article-title":"Proteolytic activation of the epithelial sodium channel (ENaC) by factor-VII-activating protease (FSAP) and its relevance for sodium retention in nephrotic mice","volume":"474","author":"Artunc","year":"2022","journal-title":"Pflugers. Arch."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Uslu, O., Herold, J., and Kanse, S.M. (2019). VEGF-A-Cleavage by FSAP and Inhibition of Neo-Vascularization. Cells, 8.","DOI":"10.3390\/cells8111396"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"483","DOI":"10.1161\/ATVBAHA.109.200451","article-title":"Hyaluronic Acid binding protein 2 is a novel regulator of vascular integrity","volume":"30","author":"Mambetsariev","year":"2010","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"18990","DOI":"10.1038\/s41598-019-55531-x","article-title":"Characterization of the enzymatic activity of the serine protease domain of Factor-VII-activating protease (FSAP)","volume":"9","author":"Nielsen","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"564","DOI":"10.1038\/nrm.2016.80","article-title":"Tight junctions: From simple barriers to multifunctional molecular gates","volume":"17","author":"Zihni","year":"2016","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1154","DOI":"10.1681\/ASN.2021070997","article-title":"Tumor Lysis Syndrome and AKI: Beyond Crystal Mechanisms","volume":"33","author":"Arnaud","year":"2022","journal-title":"J. Am. Soc. Nephrol"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"965","DOI":"10.1111\/ejn.12830","article-title":"Deficiency of Factor-VII-activating protease alters the outcome of ischemic stroke in mice","volume":"41","author":"Joshi","year":"2015","journal-title":"Eur. J. Neurosci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"e22564","DOI":"10.1096\/fj.202200828R","article-title":"Factor-VII-activating protease (FSAP) inhibits the outcome of ischemic stroke in mouse models","volume":"36","author":"Kim","year":"2022","journal-title":"FASEB J."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"e227310","DOI":"10.21769\/BioProtoc.2273","article-title":"In vitro Assays for Measuring Endothelial Permeability by Transwells and Electrical Impedance Systems","volume":"7","author":"Chen","year":"2017","journal-title":"Bio. Protocol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1111\/jth.13206","article-title":"Effect of hypoxia on tissue factor pathway inhibitor expression in breast cancer","volume":"14","author":"Cui","year":"2016","journal-title":"J. Thromb. Haemost."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/22\/13706\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:12:27Z","timestamp":1760145147000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/22\/13706"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,11,8]]},"references-count":28,"journal-issue":{"issue":"22","published-online":{"date-parts":[[2022,11]]}},"alternative-id":["ijms232213706"],"URL":"https:\/\/doi.org\/10.3390\/ijms232213706","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,11,8]]}}}