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Inhibition of protein disulfide isomerase with PACMA-31 regulates monocyte tissue factor through transcriptional and posttranscriptional mechanisms

  • Lennart Beckmann
    Affiliations
    II. Medizinische Klinik und Poliklinik, Hubertus Wald Tumorzentrum – Universitäres Cancer Center Hamburg (UCCH), Universitätsklinikum Eppendorf, Hamburg, Germany
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  • Jonathan Mäder
    Affiliations
    II. Medizinische Klinik und Poliklinik, Hubertus Wald Tumorzentrum – Universitäres Cancer Center Hamburg (UCCH), Universitätsklinikum Eppendorf, Hamburg, Germany
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  • Minna Voigtlaender
    Affiliations
    II. Medizinische Klinik und Poliklinik, Hubertus Wald Tumorzentrum – Universitäres Cancer Center Hamburg (UCCH), Universitätsklinikum Eppendorf, Hamburg, Germany
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  • Felix Klingler
    Affiliations
    II. Medizinische Klinik und Poliklinik, Hubertus Wald Tumorzentrum – Universitäres Cancer Center Hamburg (UCCH), Universitätsklinikum Eppendorf, Hamburg, Germany
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  • Anita Schulenkorf
    Affiliations
    II. Medizinische Klinik und Poliklinik, Hubertus Wald Tumorzentrum – Universitäres Cancer Center Hamburg (UCCH), Universitätsklinikum Eppendorf, Hamburg, Germany
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  • Carina Lehr
    Affiliations
    II. Medizinische Klinik und Poliklinik, Hubertus Wald Tumorzentrum – Universitäres Cancer Center Hamburg (UCCH), Universitätsklinikum Eppendorf, Hamburg, Germany
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  • Judith Regenhardt
    Affiliations
    II. Medizinische Klinik und Poliklinik, Hubertus Wald Tumorzentrum – Universitäres Cancer Center Hamburg (UCCH), Universitätsklinikum Eppendorf, Hamburg, Germany
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  • Carsten Bokemeyer
    Affiliations
    II. Medizinische Klinik und Poliklinik, Hubertus Wald Tumorzentrum – Universitäres Cancer Center Hamburg (UCCH), Universitätsklinikum Eppendorf, Hamburg, Germany
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  • Wolfram Ruf
    Affiliations
    Centrum für Thrombose und Hämostase, Universitätsmedizin der Johannes-Gutenberg-Universität Mainz, Mainz, Germany

    Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, CA, United States
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  • Author Footnotes
    1 These authors share senior authorship.
    Christina Rolling
    Footnotes
    1 These authors share senior authorship.
    Affiliations
    II. Medizinische Klinik und Poliklinik, Hubertus Wald Tumorzentrum – Universitäres Cancer Center Hamburg (UCCH), Universitätsklinikum Eppendorf, Hamburg, Germany

    Department of Cardiology, New York University School of Medicine, New York, NY, United States
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  • Author Footnotes
    1 These authors share senior authorship.
    Florian Langer
    Correspondence
    Corresponding author at: II. Medizinische Klinik und Poliklinik, Hubertus Wald Tumorzentrum - Universitäres Cancer Center Hamburg (UCCH), Universitätsklinikum Eppendorf, Martinistr. 52, D-20246 Hamburg, Germany.
    Footnotes
    1 These authors share senior authorship.
    Affiliations
    II. Medizinische Klinik und Poliklinik, Hubertus Wald Tumorzentrum – Universitäres Cancer Center Hamburg (UCCH), Universitätsklinikum Eppendorf, Hamburg, Germany
    Search for articles by this author
  • Author Footnotes
    1 These authors share senior authorship.
Published:September 30, 2022DOI:https://doi.org/10.1016/j.thromres.2022.09.024

      Highlights

      • Protein disulfide isomerase (PDI) contributes to TF regulation in monocytes.
      • Propynoic acid carbamoyl methyl-amide-31 (PACMA) is a potent PDI inhibitor.
      • PACMA regulates monocyte TF by transcriptional and posttranscriptional mechanisms.
      • PACMA can convert preformed procoagulant TF into its cryptic state.
      • PACMA amplifies LPS-induced monocyte TF production in a PAR2-dependent manner.

      Abstract

      Introduction

      Protein disulfide isomerase (PDI) contributes to tissue factor (TF) regulation in monocytes. While bacitracin and quercetin-3-rutinoside mitigate myeloid TF production, the effect of PACMA-31, a more specific PDI inhibitor with distinct pharmacologic properties, remains unclear.

      Materials and methods

      Lipopolysaccharide (LPS) stimulation of peripheral blood mononuclear cells (PBMCs) or citrate-anticoagulated whole blood was carried out in the presence of PACMA-31 or DMSO vehicle before monocytes were analyzed for TF expression, including antigen, procoagulant activity (PCA) and mRNA, release of IL-6 and TNFα, and LPS-induced signaling pathways.

      Results

      While PACMA-31 alone had no effect, coincubation with LPS and PACMA-31 (25 μM) enhanced LPS-induced monocyte TF production in whole blood. The effect was at least partially regulated on the transcriptional level and could not be explained by increased phosphatidylserine membrane exposure. In contrast, the same PACMA-31 concentrations were cytotoxic in isolated PBMCs. A lower dose of PACMA-31, however, restored the stimulating effect by enhancing IκB-NFκB signaling that also increased the release of IL-6 and TNFα. The protease-activated receptor 2 (PAR2) inhibitor ENMD547 but not TF antibody 10H10 or factor Xa inhibitor rivaroxaban prevented the stimulatory effect of PACMA-31 on inflammatory monocytes. In sharp contrast, short time incubation of LPS-stimulated PBMCs with 25 μM PACMA-31 was non-cytotoxic and significantly inhibited cellular TF PCA but not surface antigen expression.

      Conclusions

      PACMA-31 regulates monocyte TF in a concentration-dependent manner by opposing transcriptional and posttranscriptional mechanisms. While low concentrations of PACMA-31 augment monocyte TF production by amplifying LPS-dependent PAR2 signaling, high concentrations convert monocyte TF into its non-coagulant state.

      Graphical abstract

      Keywords

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      References

        • Hatahet F.
        • Ruddock L.W.
        Protein disulfide isomerase: a critical evaluation of its function in disulfide bond formation.
        Antioxid. Redox Signal. 2009; 11: 2807-2850https://doi.org/10.1089/ars.2009.2466
        • Bekendam R.H.
        • Flaumenhaft R.
        Inhibition of protein disulfide isomerase in thrombosis.
        Basic Clin. Pharmacol. Toxicol. 2016; 119: 42-48https://doi.org/10.1111/bcpt.12573
        • Lee E.
        • Lee D.H.
        Emerging roles of protein disulfide isomerase in cancer.
        BMB Rep. 2017; 50: 401-410https://doi.org/10.5483/bmbrep.2017.50.8.107
        • Uehara T.
        • Nakamura T.
        • Yao D.
        • Shi Z.-Q.
        • Gu Z.
        • Ma Y.
        • Masliah E.
        • Nomura Y.
        • Lipton S.A.
        S-nitrosylated protein-disulphide isomerase links protein misfolding to neurodegeneration.
        Nature. 2006; 441: 513-517https://doi.org/10.1038/nature04782
        • Unterberger U.
        • Höftberger R.
        • Gelpi E.
        • Flicker H.
        • Budka H.
        • Voigtländer T.
        Endoplasmic reticulum stress features are prominent in alzheimer disease but not in prion diseases in vivo.
        J. Neuropathol. Exp. Neurol. 2006; 65: 348-357https://doi.org/10.1097/01.jnen.0000218445.30535.6f
        • Xiong B.
        • Jha V.
        • Min J.-K.
        • Cho J.
        Protein disulfide isomerase in cardiovascular disease.
        Exp. Mol. Med. 2020; 52: 390-399https://doi.org/10.1038/s12276-020-0401-5
        • Cho J.
        • Furie B.C.
        • Coughlin S.R.
        • Furie B.
        A critical role for extracellular protein disulfide isomerase during thrombus formation in mice.
        J. Clin. Invest. 2008; 118: 1123-1131https://doi.org/10.1172/JCI34134
        • Sharda A.
        • Furie B.
        Regulatory role of thiol isomerases in thrombus formation.
        Expert. Rev. Hematol. 2018; 11: 437-448https://doi.org/10.1080/17474086.2018.1452612
        • Jasuja R.
        • Furie B.
        • Furie B.C.
        Endothelium-derived but not platelet-derived protein disulfide isomerase is required for thrombus formation in vivo.
        Blood. 2010; 116: 4665-4674https://doi.org/10.1182/blood-2010-04-278184
        • Cho J.
        • Kennedy D.R.
        • Lin L.
        • Huang M.
        • Merrill-Skoloff G.
        • Furie B.C.
        • Furie B.
        Protein disulfide isomerase capture during thrombus formation in vivo depends on the presence of β3 integrins.
        Blood. 2012; 120: 647-655https://doi.org/10.1182/blood-2011-08-372532
        • Reinhardt C.
        • von Brühl M.-L.
        • Manukyan D.
        • Grahl L.
        • Lorenz M.
        • Altmann B.
        • Dlugai S.
        • Hess S.
        • Konrad I.
        • Orschiedt L.
        • Mackman N.
        • Ruddock L.
        • Massberg S.
        • Engelmann B.
        Protein disulfide isomerase acts as an injury response signal that enhances fibrin generation via tissue factor activation.
        J. Clin. Invest. 2008; 118: 1110-1122https://doi.org/10.1172/JCI32376
        • Langer F.
        • Spath B.
        • Fischer C.
        • Stolz M.
        • Ayuk F.A.
        • Kröger N.
        • Bokemeyer C.
        • Ruf W.
        Rapid activation of monocyte tissue factor by antithymocyte globulin is dependent on complement and protein disulfide isomerase.
        Blood. 2013; 121: 2324-2335https://doi.org/10.1182/blood-2012-10-460493
        • Zhou J.
        • Wu Y.
        • Wang L.
        • Rauova L.
        • Hayes V.M.
        • Poncz M.
        • Essex D.W.
        The C-terminal CGHC motif of protein disulfide isomerase supports thrombosis.
        J. Clin. Invest. 2015; 125: 4391-4406https://doi.org/10.1172/JCI80319
        • Rothmeier A.S.
        • Marchese P.
        • Langer F.
        • Kamikubo Y.
        • Schaffner F.
        • Cantor J.
        • Ginsberg M.H.
        • Ruggeri Z.M.
        • Ruf W.
        Tissue factor prothrombotic activity is regulated by integrin-arf6 trafficking.
        Arterioscler. Thromb. Vasc. Biol. 2017; 37: 1323-1331https://doi.org/10.1161/ATVBAHA.117.309315
        • Prado G.N.
        • Romero J.R.
        • Rivera A.
        Endothelin-1 receptor antagonists regulate cell surface-associated protein disulfide isomerase in sickle cell disease.
        FASEB J. 2013; 27: 4619-4629https://doi.org/10.1096/fj.13-228577
        • Jasuja R.
        • Passam F.H.
        • Kennedy D.R.
        • Kim S.H.
        • van Hessem L.
        • Lin L.
        • Bowley S.R.
        • Joshi S.S.
        • Dilks J.R.
        • Furie B.
        • Furie B.C.
        • Flaumenhaft R.
        Protein disulfide isomerase inhibitors constitute a new class of antithrombotic agents.
        J. Clin. Invest. 2012; 122: 2104-2113https://doi.org/10.1172/JCI61228
        • Stopa J.D.
        • Neuberg D.
        • Puligandla M.
        • Furie B.
        • Flaumenhaft R.
        • Zwicker J.I.
        Protein disulfide isomerase inhibition blocks thrombin generation in humans by interfering with platelet factor V activation.
        JCI Insight. 2017; 2e89373https://doi.org/10.1172/jci.insight.89373
        • Zwicker J.I.
        • Schlechter B.L.
        • Stopa J.D.
        • Liebman H.A.
        • Aggarwal A.
        • Puligandla M.
        • Caughey T.
        • Bauer K.A.
        • Kuemmerle N.
        • Wong E.
        • Wun T.
        • McLaughlin M.
        • Hidalgo M.
        • Neuberg D.
        • Furie B.
        • Flaumenhaft R.
        Targeting protein disulfide isomerase with the flavonoid isoquercetin to improve hypercoagulability in advanced cancer.
        JCI Insight. 2019; 4https://doi.org/10.1172/jci.insight.125851
        • Østerud B.
        Tissue factor expression in blood cells.
        Thromb. Res. 2010; 125: S31-S34https://doi.org/10.1016/j.thromres.2010.01.032
        • Shantsila E.
        • Lip G.Y.H.
        The role of monocytes in thrombotic disorders. Insights from tissue factor, monocyte-platelet aggregates and novel mechanisms.
        Thromb. Haemost. 2009; 102: 916-924https://doi.org/10.1160/TH09-01-0023
        • Grover S.P.
        • Mackman N.
        Tissue factor: an essential mediator of hemostasis and trigger of thrombosis.
        Arterioscler. Thromb. Vasc. Biol. 2018; 38: 709-725https://doi.org/10.1161/ATVBAHA.117.309846
        • Zelaya H.
        • Rothmeier A.S.
        • Ruf W.
        Tissue factor at the crossroad of coagulation and cell signaling.
        J. Thromb. Haemost. 2018; 16: 1941-1952https://doi.org/10.1111/jth.14246
        • Hisada Y.
        • Mackman N.
        Tissue factor and cancer: regulation, tumor growth, and metastasis.
        Semin. Thromb. Hemost. 2019; 45: 385-395https://doi.org/10.1055/s-0039-1687894
        • Langer F.
        • Ruf W.
        Synergies of phosphatidylserine and protein disulfide isomerase in tissue factor activation.
        Thromb. Haemost. 2014; 111: 590-597https://doi.org/10.1160/TH13-09-0802
        • Ansari S.A.
        • Pendurthi U.R.
        • Rao L.V.M.
        Role of cell surface lipids and thiol-disulphide exchange pathways in regulating the encryption and decryption of tissue factor.
        Thromb. Haemost. 2019; 119: 860-870https://doi.org/10.1055/s-0039-1681102
        • Drake T.A.
        • Ruf W.
        • Morrissey J.H.
        • Edgington T.S.
        Functional tissue factor is entirely cell surface expressed on lipopolysaccharide-stimulated human blood monocytes and a constitutively tissue factor-producing neoplastic cell line.
        J. Cell Biol. 1989; 109: 389-395https://doi.org/10.1083/jcb.109.1.389
        • Beckmann L.
        • Rolling C.C.
        • Voigtländer M.
        • Mäder J.
        • Klingler F.
        • Schulenkorf A.
        • Lehr C.
        • Bokemeyer C.
        • Ruf W.
        • Langer F.
        Bacitracin and rutin regulate tissue factor production in inflammatory monocytes and acute myeloid leukemia blasts.
        Cancers (Basel). 2021; 13https://doi.org/10.3390/cancers13163941
        • Yamada R.
        • Cao X.
        • Butkevich A.N.
        • Millard M.
        • Odde S.
        • Mordwinkin N.
        • Gundla R.
        • Zandi E.
        • Louie S.G.
        • Petasis N.A.
        • Neamati N.
        Discovery and preclinical evaluation of a novel class of cytotoxic propynoic acid carbamoyl methyl amides (PACMAs).
        J. Med. Chem. 2011; 54: 2902-2914https://doi.org/10.1021/jm101655d
        • Xu S.
        • Butkevich A.N.
        • Yamada R.
        • Zhou Y.
        • Debnath B.
        • Duncan R.
        • Zandi E.
        • Petasis N.A.
        • Neamati N.
        Discovery of an orally active small-molecule irreversible inhibitor of protein disulfide isomerase for ovarian cancer treatment.
        Proc. Natl. Acad. Sci. U. S. A. 2012; 109: 16348-16353https://doi.org/10.1073/pnas.1205226109
        • Lin L.
        • Gopal S.
        • Sharda A.
        • Passam F.
        • Bowley S.R.
        • Stopa J.
        • Xue G.
        • Yuan C.
        • Furie B.C.
        • Flaumenhaft R.
        • Huang M.
        • Furie B.
        Quercetin-3-rutinoside inhibits protein disulfide isomerase by binding to its b'x domain.
        J. Biol. Chem. 2015; 290: 23543-23552https://doi.org/10.1074/jbc.M115.666180
        • Popielarski M.
        • Ponamarczuk H.
        • Stasiak M.
        • Watała C.
        • Świątkowska M.
        Modifications of disulfide bonds in breast cancer cell migration and invasiveness.
        Am. J. Cancer Res. 2019; 9: 1554-1582
        • Vatolin S.
        • Phillips J.G.
        • Jha B.K.
        • Govindgari S.
        • Hu J.
        • Grabowski D.
        • Parker Y.
        • Lindner D.J.
        • Zhong F.
        • Distelhorst C.W.
        • Smith M.R.
        • Cotta C.
        • Xu Y.
        • Chilakala S.
        • Kuang R.R.
        • Tall S.
        • Reu F.J.
        Novel protein disulfide isomerase inhibitor with anticancer activity in multiple myeloma.
        Cancer Res. 2016; 76: 3340-3350https://doi.org/10.1158/0008-5472.CAN-15-3099
        • Won J.-K.
        • Yu S.J.
        • Hwang C.Y.
        • Cho S.-H.
        • Park S.-M.
        • Kim K.
        • Choi W.-M.
        • Cho H.
        • Cho E.J.
        • Lee J.-H.
        • Lee K.B.
        • Kim Y.J.
        • Suh K.-S.
        • Jang J.-J.
        • Kim C.Y.
        • Yoon J.-H.
        • Cho K.-H.
        Protein disulfide isomerase inhibition synergistically enhances the efficacy of sorafenib for hepatocellular carcinoma.
        Hepatology. 2017; 66: 855-868https://doi.org/10.1002/hep.29237
        • Kelso E.B.
        • Lockhart J.C.
        • Hembrough T.
        • Dunning L.
        • Plevin R.
        • Hollenberg M.D.
        • Sommerhoff C.P.
        • McLean J.S.
        • Ferrell W.R.
        Therapeutic promise of proteinase-activated receptor-2 antagonism in joint inflammation.
        J. Pharmacol. Exp. Ther. 2006; 316: 1017-1024https://doi.org/10.1124/jpet.105.093807
        • Kelso E.B.
        • Ferrell W.R.
        • Lockhart J.C.
        • Elias-Jones I.
        • Hembrough T.
        • Dunning L.
        • Gracie J.A.
        • McInnes I.B.
        Expression and proinflammatory role of proteinase-activated receptor 2 in rheumatoid synovium: ex vivo studies using a novel proteinase-activated receptor 2 antagonist.
        Arthritis Rheum. 2007; 56: 765-771https://doi.org/10.1002/art.22423
        • Kim K.
        • Hahm E.
        • Li J.
        • Holbrook L.-M.
        • Sasikumar P.
        • Stanley R.G.
        • Ushio-Fukai M.
        • Gibbins J.M.
        • Cho J.
        Platelet protein disulfide isomerase is required for thrombus formation but not for hemostasis in mice.
        Blood. 2013; 122: 1052-1061https://doi.org/10.1182/blood-2013-03-492504
        • Guha M.
        • Mackman N.
        LPS induction of gene expression in human monocytes.
        Cell. Signal. 2001; 13: 85-94https://doi.org/10.1016/s0898-6568(00)00149-2
        • Bode M.
        • Mackman N.
        Regulation of tissue factor gene expression in monocytes and endothelial cells: thromboxane A2 as a new player.
        Vasc. Pharmacol. 2014; 62: 57-62https://doi.org/10.1016/j.vph.2014.05.005
        • Badeanlou L.
        • Furlan-Freguia C.
        • Yang G.
        • Ruf W.
        • Samad F.
        Tissue factor-protease-activated receptor 2 signaling promotes diet-induced obesity and adipose inflammation.
        Nat. Med. 2011; 17: 1490-1497https://doi.org/10.1038/nm.2461
        • Rallabhandi P.
        • Nhu Q.M.
        • Toshchakov V.Y.
        • Piao W.
        • Medvedev A.E.
        • Hollenberg M.D.
        • Fasano A.
        • Vogel S.N.
        Analysis of proteinase-activated receptor 2 and TLR4 signal transduction: a novel paradigm for receptor cooperativity.
        J. Biol. Chem. 2008; 283: 24314-24325https://doi.org/10.1074/jbc.M804800200
        • Liang H.P.H.
        • Kerschen E.J.
        • Hernandez I.
        • Basu S.
        • Zogg M.
        • Botros F.
        • Jia S.
        • Hessner M.J.
        • Griffin J.H.
        • Ruf W.
        • Weiler H.
        EPCR-dependent PAR2 activation by the blood coagulation initiation complex regulates LPS-triggered interferon responses in mice.
        Blood. 2015; 125: 2845-2854https://doi.org/10.1182/blood-2014-11-610717
        • Antoniak S.
        • Mackman N.
        Multiple roles of the coagulation protease cascade during virus infection.
        Blood. 2014; 123: 2605-2613https://doi.org/10.1182/blood-2013-09-526277
        • Colognato R.
        • Slupsky J.R.
        • Jendrach M.
        • Burysek L.
        • Syrovets T.
        • Simmet T.
        Differential expression and regulation of protease-activated receptors in human peripheral monocytes and monocyte-derived antigen-presenting cells.
        Blood. 2003; 102: 2645-2652https://doi.org/10.1182/blood-2002-08-2497
        • Johansson U.
        • Lawson C.
        • Dabare M.
        • Syndercombe-Court D.
        • Newland A.C.
        • Howells G.L.
        • Macey M.G.
        Human peripheral blood monocytes express protease receptor-2 and respond to receptor activation by production of IL-6, IL-8, and IL-1{beta}.
        J. Leukoc. Biol. 2005; 78: 967-975https://doi.org/10.1189/jlb.0704422
        • Crilly A.
        • Burns E.
        • Nickdel M.B.
        • Lockhart J.C.
        • Perry M.E.
        • Ferrell P.W.
        • Baxter D.
        • Dale J.
        • Dunning L.
        • Wilson H.
        • Nijjar J.S.
        • Gracie J.A.
        • Ferrell W.R.
        • McInnes I.B.
        PAR(2) expression in peripheral blood monocytes of patients with rheumatoid arthritis.
        Ann. Rheum. Dis. 2012; 71: 1049-1054https://doi.org/10.1136/annrheumdis-2011-200703
        • Ahamed J.
        • Versteeg H.H.
        • Kerver M.
        • Chen V.M.
        • Mueller B.M.
        • Hogg P.J.
        • Ruf W.
        Disulfide isomerization switches tissue factor from coagulation to cell signaling.
        Proc. Natl. Acad. Sci. U. S. A. 2006; 103: 13932-13937https://doi.org/10.1073/pnas.0606411103
        • Versteeg H.H.
        • Schaffner F.
        • Kerver M.
        • Petersen H.H.
        • Ahamed J.
        • Felding-Habermann B.
        • Takada Y.
        • Mueller B.M.
        • Ruf W.
        Inhibition of tissue factor signaling suppresses tumor growth.
        Blood. 2008; 111: 190-199https://doi.org/10.1182/blood-2007-07-101048
        • Subramaniam S.
        • Jurk K.
        • Hobohm L.
        • Jäckel S.
        • Saffarzadeh M.
        • Schwierczek K.
        • Wenzel P.
        • Langer F.
        • Reinhardt C.
        • Ruf W.
        Distinct contributions of complement factors to platelet activation and fibrin formation in venous thrombus development.
        Blood. 2017; 129: 2291-2302https://doi.org/10.1182/blood-2016-11-749879
        • Versteeg H.H.
        • Ruf W.
        Tissue factor coagulant function is enhanced by protein-disulfide isomerase independent of oxidoreductase activity.
        J. Biol. Chem. 2007; 282: 25416-25424https://doi.org/10.1074/jbc.M702410200
        • van den Hengel Lisa G.
        • Osanto S.
        • Reitsma P.H.
        • Versteeg H.H.
        Murine tissue factor coagulant activity is critically dependent on the presence of an intact allosteric disulfide.
        Haematologica. 2013; 98: 153-158https://doi.org/10.3324/haematol.2012.069997
        • Bekendam R.H.
        • Bendapudi P.K.
        • Lin L.
        • Nag P.P.
        • Pu J.
        • Kennedy D.R.
        • Feldenzer A.
        • Chiu J.
        • Cook K.M.
        • Furie B.
        • Huang M.
        • Hogg P.J.
        • Flaumenhaft R.
        A substrate-driven allosteric switch that enhances PDI catalytic activity.
        Nat. Commun. 2016; 7: 12579https://doi.org/10.1038/ncomms12579
        • Chen V.M.
        • Ahamed J.
        • Versteeg H.H.
        • Berndt M.C.
        • Ruf W.
        • Hogg P.J.
        Evidence for activation of tissue factor by an allosteric disulfide bond.
        Biochemistry. 2006; 45: 12020-12028https://doi.org/10.1021/bi061271a
        • Chen F.
        • Zhao Z.
        • Zhou J.
        • Lu Y.
        • Essex D.W.
        • Wu Y.
        Protein disulfide isomerase enhances tissue factor-dependent thrombin generation.
        Biochem. Biophys. Res. Commun. 2018; 501: 172-177https://doi.org/10.1016/j.bbrc.2018.04.207
        • Krajewski D.
        • Polukort S.H.
        • Gelzinis J.
        • Rovatti J.
        • Kaczenski E.
        • Galinski C.
        • Pantos M.
        • Shah N.N.
        • Schneider S.S.
        • Kennedy D.R.
        • Mathias C.B.
        Protein disulfide isomerases regulate IgE-mediated mast cell responses and their inhibition confers protective effects during food allergy.
        Front. Immunol. 2020; 11606837https://doi.org/10.3389/fimmu.2020.606837
        • Higuchi T.
        • Watanabe Y.
        • Waga I.
        Protein disulfide isomerase suppresses the transcriptional activity of NF-kappaB.
        Biochem. Biophys. Res. Commun. 2004; 318: 46-52https://doi.org/10.1016/j.bbrc.2004.04.002
        • Xiao Y.
        • Li C.
        • Gu M.
        • Wang H.
        • Chen W.
        • Luo G.
        • Yang G.
        • Zhang Z.
        • Zhang Y.
        • Xian G.
        • Li Z.
        • Sheng P.
        Protein disulfide isomerase silence inhibits inflammatory functions of macrophages by suppressing reactive oxygen species and NF-κB pathway.
        Inflammation. 2018; 41: 614-625https://doi.org/10.1007/s10753-017-0717-z
        • Hahm E.
        • Li J.
        • Kim K.
        • Huh S.
        • Rogelj S.
        • Cho J.
        Extracellular protein disulfide isomerase regulates ligand-binding activity of αMβ2 integrin and neutrophil recruitment during vascular inflammation.
        Blood. 2013; 121: S1-S15https://doi.org/10.1182/blood-2012-11-467985
        • Short J.D.
        • Downs K.
        • Tavakoli S.
        • Asmis R.
        Protein thiol redox signaling in monocytes and macrophages.
        Antioxid. Redox Signal. 2016; 25: 816-835https://doi.org/10.1089/ars.2016.6697
        • Gutmann C.
        • Siow R.
        • Gwozdz A.M.
        • Saha P.
        • Smith A.
        Reactive oxygen species in venous thrombosis.
        Int. J. Mol. Sci. 2020; 21https://doi.org/10.3390/ijms21061918
        • Furlan-Freguia C.
        • Marchese P.
        • Gruber A.
        • Ruggeri Z.M.
        • Ruf W.
        P2X7 receptor signaling contributes to tissue factor-dependent thrombosis in mice.
        J. Clin. Invest. 2011; 121: 2932-2944https://doi.org/10.1172/JCI46129
        • Rothmeier A.S.
        • Marchese P.
        • Petrich B.G.
        • Furlan-Freguia C.
        • Ginsberg M.H.
        • Ruggeri Z.M.
        • Ruf W.
        Caspase-1-mediated pathway promotes generation of thromboinflammatory microparticles.
        J. Clin. Invest. 2015; 125: 1471-1484https://doi.org/10.1172/JCI79329
        • Banfi C.
        • Brioschi M.
        • Barbieri S.S.
        • Eligini S.
        • Barcella S.
        • Tremoli E.
        • Colli S.
        • Mussoni L.
        Mitochondrial reactive oxygen species: a common pathway for PAR1- and PAR2-mediated tissue factor induction in human endothelial cells.
        J. Thromb. Haemost. 2009; 7: 206-216https://doi.org/10.1111/j.1538-7836.2008.03204.x
        • Bang E.
        • Kim D.H.
        • Chung H.Y.
        Protease-activated receptor 2 induces ROS-mediated inflammation through akt-mediated NF-κB and FoxO6 modulation during skin photoaging.
        Redox Biol. 2021; 44102022https://doi.org/10.1016/j.redox.2021.102022
        • Xu Q.
        • Zhang J.
        • Zhao Z.
        • Chu Y.
        • Fang J.
        Revealing PACMA 31 as a new chemical type TrxR inhibitor to promote cancer cell apoptosis.
        Biochim. Biophys. Acta, Mol. Cell Res. 2022; 1869119323https://doi.org/10.1016/j.bbamcr.2022.119323
        • Liang H.P.H.
        • Kerschen E.J.
        • Hernandez I.
        EPCR-dependent PAR2 activation by the blood coagulation initiation complex regulates LPS-triggered interferon responses in mice.
        Blood. 2015; 125 (Blood 131 (2018) 2508): 2845-2854https://doi.org/10.1182/blood-2018-04-845123
        • Chi L.
        • Li Y.
        • Stehno-Bittel L.
        • Gao J.
        • Morrison D.C.
        • Stechschulte D.J.
        • Dileepan K.N.
        Interleukin-6 production by endothelial cells via stimulation of protease-activated receptors is amplified by endotoxin and tumor necrosis factor-alpha.
        J. Interf. Cytokine Res. 2001; 21: 231-240https://doi.org/10.1089/107999001750169871
        • Zhou B.
        • Zhou H.
        • Ling S.
        • Guo D.
        • Yan Y.
        • Zhou F.
        • Wu Y.
        Activation of PAR2 or/and TLR4 promotes SW620 cell proliferation and migration via phosphorylation of ERK1/2.
        Oncol. Rep. 2011; 25: 503-511https://doi.org/10.3892/or.2010.1077
        • Bucci M.
        • Vellecco V.
        • Harrington L.
        • Brancaleone V.
        • Roviezzo F.
        • Mattace Raso G.
        • Ianaro A.
        • Lungarella G.
        • de Palma R.
        • Meli R.
        • Cirino G.
        Cross-talk between toll-like receptor 4 (TLR4) and proteinase-activated receptor 2 (PAR(2) ) is involved in vascular function.
        Br. J. Pharmacol. 2013; 168: 411-420https://doi.org/10.1111/j.1476-5381.2012.02205.x
        • Widera D.
        • Aguilar R.Martínez
        • Cottrell G.S.
        Toll-like receptor 4 and protease-activated receptor 2 in physiology and pathophysiology of the nervous system: more than just receptor cooperation?.
        Neural Regen. Res. 2019; 14: 1196-1201https://doi.org/10.4103/1673-5374.251290
        • Rothmeier A.S.
        • Ruf W.
        Protease-activated receptor 2 signaling in inflammation.
        Semin. Immunopathol. 2012; 34: 133-149https://doi.org/10.1007/s00281-011-0289-1
        • Müller-Calleja N.
        • Hollerbach A.
        • Ritter S.
        • Pedrosa D.G.
        • Strand D.
        • Graf C.
        • Reinhardt C.
        • Strand S.
        • Poncelet P.
        • Griffin J.H.
        • Lackner K.J.
        • Ruf W.
        Tissue factor pathway inhibitor primes monocytes for antiphospholipid antibody-induced thrombosis.
        Blood. 2019; 134: 1119-1131https://doi.org/10.1182/blood.2019001530
        • Rothmeier A.S.
        • Liu E.
        • Chakrabarty S.
        • Disse J.
        • Mueller B.M.
        • Østergaard H.
        • Ruf W.
        Identification of the integrin-binding site on coagulation factor VIIa required for proangiogenic PAR2 signaling.
        Blood. 2018; 131: 674-685https://doi.org/10.1182/blood-2017-02-768218
        • Disse J.
        • Petersen H.H.
        • Larsen K.S.
        • Persson E.
        • Esmon N.
        • Esmon C.T.
        • Teyton L.
        • Petersen L.C.
        • Ruf W.
        The endothelial protein C receptor supports tissue factor ternary coagulation initiation complex signaling through protease-activated receptors *.
        J. Biol. Chem. 2011; 286: 5756-5767https://doi.org/10.1074/jbc.M110.201228
        • Ruf W.
        Roles of factor xa beyond coagulation.
        J. Thromb. Thrombolysis. 2021; 52: 391-396https://doi.org/10.1007/s11239-021-02458-8
        • Graf C.
        • Wilgenbus P.
        • Pagel S.
        • Pott J.
        • Marini F.
        • Reyda S.
        • Kitano M.
        • Macher-Göppinger S.
        • Weiler H.
        • Ruf W.
        Myeloid cell-synthesized coagulation factor X dampens antitumor immunity.
        Sci. Immunol. 2019; 4https://doi.org/10.1126/sciimmunol.aaw8405
        • Müller-Calleja N.
        • Hollerbach A.
        • Royce J.
        • Ritter S.
        • Pedrosa D.
        • Madhusudhan T.
        • Teifel S.
        • Meineck M.
        • Häuser F.
        • Canisius A.
        • Nguyen T.S.
        • Braun J.
        • Bruns K.
        • Etzold A.
        • Zechner U.
        • Strand S.
        • Radsak M.
        • Strand D.
        • Gu J.-M.
        • Weinmann-Menke J.
        • Esmon C.T.
        • Teyton L.
        • Lackner K.J.
        • Ruf W.
        Lipid presentation by the protein C receptor links coagulation with autoimmunity.
        Science. 2021; 371https://doi.org/10.1126/science.abc0956
        • Rosenbaum D.M.
        • Rasmussen S.G.F.
        • Kobilka B.K.
        The structure and function of G-protein-coupled receptors.
        Nature. 2009; 459: 356-363https://doi.org/10.1038/nature08144