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A missense mutation in lectin domain of thrombomodulin causing functional deficiency

  • Author Footnotes
    # Ma Jiewen and Tao Yanyi are co-first authors.
    Ma Jiewen
    Footnotes
    # Ma Jiewen and Tao Yanyi are co-first authors.
    Affiliations
    Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
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  • Author Footnotes
    # Ma Jiewen and Tao Yanyi are co-first authors.
    Tao yanyi
    Footnotes
    # Ma Jiewen and Tao Yanyi are co-first authors.
    Affiliations
    Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
    Search for articles by this author
  • Feng Yuanzheng
    Affiliations
    Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
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  • Cheng Zhipeng
    Affiliations
    Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
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  • Lin Wenyi
    Affiliations
    Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
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  • Hu Bei
    Affiliations
    Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
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  • Hu Yu
    Correspondence
    Reprint requests: Tang Liang, Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, No. 1277 Jiefang Avenue, Wuhan, 430022, China
    Affiliations
    Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
    Search for articles by this author
  • Liang V. Tang
    Correspondence
    Reprint requests: Tang Liang, Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, No. 1277 Jiefang Avenue, Wuhan, 430022, China
    Affiliations
    Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
    Search for articles by this author
  • Author Footnotes
    # Ma Jiewen and Tao Yanyi are co-first authors.

      Abstract

      Thrombomodulin (TM) functions in coagulation, fibrinolysis and inflammation by its cofactor activity for protein C, thrombin-activatable fibrinolysis inhibitor (TAFI) activation and high mobility group box 1 (HMGB1) degradation induced by thrombin. It has been widely reported that mutations in TM are related to thromboembolic diseases but hardly in lectin domain. Here we report our findings about the functional deficiencies in TM caused by substitution of aspartate with tyrosine at residue 126. Three patients suffering from recurrent thromboembolic diseases were identified with this mutation and their plasma soluble TM levels were decreased. Transfected cells expressing wild-type TM or the variant and corresponding proteins were used to examine TM functions in vitro. The cofactor activity of the mutant for protein C, TAFI activation was reduced to approximately 50% and 60% respectively. Loss in anti-inflammation due to weakened HMGB1 degradation was also observed. And the study with thrombosis models of mice suggested the decreased inhibition of thrombus development of the mutant. Together the results showed deleterious changes on TM function caused by this mutation, which may explain the thrombophilia tendency of the patients. This work provided supportive evidence that mutation in lectin domain of TM might be related to thrombotic diseases and may help us better understand the physiological roles of TM.

      Abbreviations:

      TM (thrombomodulin), EGF (epidermal growth factor), Ser/Thr (serine/threonine), sTM (soluble thrombomodulin), PC (protein C), APC (activated protein C), TAFI (thrombin-activatable fibrinolysis inhibitor), TAFIa (activated TAFI), tPA (tissue-type plasminogen activator), HMGB1 (high mobility group box 1), Hip-Arg (hippuryl-L-arginine), Asp/D (aspartate), Tyr/Y (tyrosine), PC/PS/PE (L-a-phosphatidylcholine/L-a-phosphatidylserine/L-a-phosphatidylethanolamine), PPACK (H-D-Phe-Pro-Arg-chloromethylketone), RT-PCR (real-time reverse-transcription polymerase chain reaction), SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis), SPR (Surface Plasmon Resonance), PBS (phosphate-buffered saline), TF (tissue factor), IVC (inferior vena cava), KD (affinity constant), MFI (median fluorescence intensity)
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      References

        • Weiler H
        • Isermann BH.
        Thrombomodulin.
        J Thromb Haemost. 2003; 1: 1515-1524
        • Sadler JE.
        Thrombomodulin structure and function.
        Thromb Haemost. 1997; 78: 392-395
        • Esmon CT
        • Owen WG.
        Identification of an endothelial cell cofactor for thrombin-catalyzed activation of protein C.
        Proc Natl Acad Sci U S A. 1981; 78: 2249-2252
        • Fuentes-Prior P
        • Iwanaga Y
        • Huber R
        • et al.
        Structural basis for the anticoagulant activity of the thrombin-thrombomodulin complex.
        Nature. 2000; 404: 518-525
        • Dittman WA
        • Majerus PW.
        Structure and function of thrombomodulin: a natural anticoagulant.
        Blood. 1990; 75: 329-336
        • Ishii H
        • Majerus PW.
        Thrombomodulin is present in human plasma and urine.
        J Clin Invest. 1985; 76: 2178-2181
        • Suzuki K
        • Stenflo J
        • Dahlbäck B
        • Teodorsson B.
        Inactivation of human coagulation factor V by activated protein C.
        J Biol Chem. 1983; 258: 1914-1920
        • Rezaie AR.
        Regulation of the protein C anticoagulant and antiinflammatory pathways.
        Curr Med Chem. 2010; 17: 2059-2069
        • Tsiang M
        • Lentz SR
        • Sadler JE.
        Functional domains of membrane-bound human thrombomodulin. EGF-like domains four to six and the serine/threonine-rich domain are required for cofactor activity.
        J Biol Chem. 1992; 267: 6164-6170
        • Kokame K
        • Zheng X
        • Sadler JE.
        Activation of thrombin-activable fibrinolysis inhibitor requires epidermal growth factor-like domain 3 of thrombomodulin and is inhibited competitively by protein C.
        J Biol Chem. 1998; 273: 12135-12139
        • Loghmani H
        • Conway EM.
        Exploring traditional and nontraditional roles for thrombomodulin.
        Blood. 2018; 132: 148-158
        • Bajzar L
        • Morser J
        • Nesheim M.
        TAFI, or plasma procarboxypeptidase B, couples the coagulation and fibrinolytic cascades through the thrombin-thrombomodulin complex.
        J Biol Chem. 1996; 271: 16603-16608
        • Ito T
        • Kawahara K
        • Okamoto K
        • et al.
        Proteolytic cleavage of high mobility group box 1 protein by thrombin-thrombomodulin complexes.
        Arterioscler Thromb Vasc Biol. 2008; 28: 1825-1830
        • Conway EM.
        Thrombomodulin and its role in inflammation.
        Semin Immunopathol. 2012; 34: 107-125
        • Li YH
        • Kuo CH
        • Shi GY
        • Wu HL.
        The role of thrombomodulin lectin-like domain in inflammation.
        J Biomed Sci. 2012; 19: 34
        • Kunz G
        • Ohlin AK
        • Adami A
        • Zöller B
        • Svensson P
        • Lane DA
        Naturally occurring mutations in the thrombomodulin gene leading to impaired expression and function.
        Blood. 2002; 99: 3646-3653
        • Tang L
        • Wang HF
        • Lu X
        • et al.
        Common genetic risk factors for venous thrombosis in the Chinese population.
        Am J Hum Genet. 2013; 92: 177-187
        • Mattsson C
        • Menschik-Lundin A
        • Nylander S
        • Gyzander E
        • Deinum J.
        Effect of different types of thrombin inhibitors on thrombin/thrombomodulin modulated activation of protein C in vitro.
        Thromb Res. 2001; 104: 475-486
        • Mosnier LO
        • von Dem BP
        • Meijers JC
        • Bouma BN
        Plasma TAFI levels influence the clot lysis time in healthy individuals in the presence of an intact intrinsic pathway of coagulation.
        Thromb Haemost. 1998; 80: 829-835
        • Subramaniam S
        • Kanse SM.
        Ferric chloride-induced arterial thrombosis in mice.
        Curr Protoc Mouse Biol. 2014; 4: 151-164
        • Plug T
        • Kramer G
        • Meijers JC.
        A role for arginine-12 in thrombin-thrombomodulin-mediated activation of thrombin-activatable fibrinolysis inhibitor.
        J Thromb Haemost. 2014; 12: 1717-1725
        • Weiler H.
        Mouse models of thrombosis: thrombomodulin.
        Thromb Haemost. 2004; 92: 467-477
        • Ohlin AK
        • Marlar RA.
        The first mutation identified in the thrombomodulin gene in a 45-year-old man presenting with thromboembolic disease.
        Blood. 1995; 85: 330-336
        • Nakazawa F
        • Koyama T
        • Saito T
        • et al.
        Thrombomodulin with the Asp468Tyr mutation is expressed on the cell surface with normal cofactor activity for protein C activation.
        Br J Haematol. 1999; 106: 416-420
        • Ohlin AK
        • Marlar RA.
        Thrombomodulin gene defects in families with thromboembolic disease–a report on four families.
        Thromb Haemost. 1999; 81: 338-344
        • Ohlin AK
        • Norlund L
        • Marlar RA.
        Thrombomodulin gene variations and thromboembolic disease.
        Thromb Haemost. 1997; 78: 396-400
        • Langdown J
        • Luddington RJ
        • Huntington JA
        • Baglin TP.
        A hereditary bleeding disorder resulting from a premature stop codon in thrombomodulin.
        Blood. 2014; 124 (p.Cys537Stop): 1951-1956
        • Westbury SK
        • Whyte CS
        • Stephens J
        • et al.
        A new pedigree with thrombomodulin-associated coagulopathy in which delayed fibrinolysis is partially attenuated by co-inherited TAFI deficiency.
        J Thromb Haemost. 2020; 18: 2209-2214
        • Burley K
        • Whyte CS
        • Westbury SK
        • et al.
        Altered fibrinolysis in autosomal dominant thrombomodulin-associated coagulopathy.
        Blood. 2016; 128: 1879-1883
        • Okada M
        • Tominaga N
        • Honda G
        • et al.
        A case of thrombomodulin mutation causing defective thrombin binding with absence of protein C and TAFI activation.
        Blood Adv. 2020; 4: 2631-2639
        • Kager LM
        • Wiersinga WJ
        • Roelofs JJ
        • et al.
        Mice lacking the lectin-like domain of thrombomodulin are protected against melioidosis.
        Crit Care Med. 2014; 42: e221-e230
        • Szilágyi A
        • Kiss N
        • Bereczki C
        • et al.
        The role of complement in Streptococcus pneumoniae-associated haemolytic uraemic syndrome.
        Nephrol Dial Transplant. 2013; 28: 2237-2245
        • Maga TK
        • Nishimura CJ
        • Weaver AE
        • Frees KL
        • Smith RJ.
        Mutations in alternative pathway complement proteins in American patients with atypical hemolytic uremic syndrome.
        Hum Mutat. 2010; 31: E1445-E1460
        • Delvaeye M
        • Noris M
        • De Vriese A
        • et al.
        Thrombomodulin mutations in atypical hemolytic-uremic syndrome.
        N Engl J Med. 2009; 361: 345-357
        • Doggen CJ
        • Kunz G
        • Rosendaal FR
        • et al.
        A mutation in the thrombomodulin gene, 127G to A coding for Ala25Thr, and the risk of myocardial infarction in men.
        Thromb Haemost. 1998; 80: 743-748
        • Bae JS
        • Rezaie AR.
        Activated protein C inhibits high mobility group box 1 signaling in endothelial cells.
        Blood. 2011; 118: 3952-3959
        • Salomaa V
        • Matei C
        • Aleksic N
        • et al.
        Soluble thrombomodulin as a predictor of incident coronary heart disease and symptomless carotid artery atherosclerosis in the Atherosclerosis Risk in Communities (ARIC) Study: a case-cohort study.
        Lancet. 1999; 353: 1729-1734
        • Kudo D
        • Goto T
        • Uchimido R
        • et al.
        Coagulation phenotypes in sepsis and effects of recombinant human thrombomodulin: an analysis of three multicentre observational studies.
        Crit Care. 2021; 25: 114