Molecular Medicine

, Volume 21, Issue 1, pp 355–363 | Cite as

Preserved Expression of mRNA Coding von Willebrand Factor-Cleaving Protease ADAMTS13 by Selenite and Activated Protein C

  • Michael L. Ekaney
  • Clemens L. Bockmeyer
  • Maik Sossdorf
  • Philipp A. Reuken
  • Florian Conradi
  • Tobias Schuerholz
  • Markus F. Blaess
  • Scott L. Friedman
  • Wolfgang Lösche
  • Michael Bauer
  • Ralf A. Claus
Research Article


In sepsis, the severity-dependent decrease of von Willebrand factor (VWF)-inactivating protease, a disintegrin and metalloproteinase with thrombospondin motifs 13 (ADAMTS13), results in platelet aggregation and consumption, leading to sepsis-associated thrombotic microangiopathy (TMA) and organ failure. Previous reports assessing its functional deficiency have pinpointed involvement of autoantibodies or mutations to propagate thrombotic thrombocytopenic purpura (TTP). However, mechanisms of acquired ADAMTS13 deficiency during host response remain unclear. To enhance understanding of ADAMTS13 deficiency in sepsis, we evaluated changes in expression of mRNA coding ADAMTS13 during septic conditions using primary cellular sources of the protease. We hypothesized that proinflammatory cytokines and constituents of serum from septic patients affect the transcriptional level of ADAMTS13 in vitro, and previously recommended therapeutic agents as adjunctive therapy for sepsis interact therewith. Cultured hepatic stellate cells (HSCs), endothelial cells (HMEC) and human precision-cut liver slices as an ex vivo model were stimulated with sepsis prototypic cytokines, bacterial endotoxin and pooled serum obtained from septic patients. Stimulation resulted in a significant decrease in ADAMTS13 mRNA between 10% and 80% of basal transcriptional rates. Costimulation of selenite or recombinant activated protein C (APC) with serum prevented ADAMTS13 decrease in HSCs and increased ADAMTS13 transcripts in HMEC. In archived clinical samples, the activity of ADAMTS13 in septic patients treated with APC (n = 5) increased with an accompanying decrease in VWF propeptide as surrogate for improved endothelial function. In conclusion, proinflammatory conditions of sepsis repress mRNA coding ADAMTS13 and the ameliorating effect by selenite and APC may support the concept for identification of beneficial mechanisms triggered by these drugs at a molecular level.



The authors thank Gordon Philipp Otto, Sina Coldewey, Brigitte Specht, Barbara Schmidt and Edith Walther for their technical support and helpful comments. The authors acknowledge Utz Settmacher for providing surgical waste material and Dieter Muller for assistance in handling HPLS. This study was supported by grants from the German Research Foundation to RA Claus (DFG CL 173/4–1); German Federal Ministry of Education and Research within the Center for Sepsis Control and Care (grant 01 EO 1002, Project D1.9; PhD fellowship to ML Ekaney); and by the United States National Institutes of Health (DK56621 to SL Friedman).


  1. 1.
    Angus DC, Pereira CA, Silva E. (2006) Epidemiology of severe sepsis around the world. Endocr. Metab. Immune Disord. Drug Targets 6:207–12.CrossRefGoogle Scholar
  2. 2.
    Martin GS, Mannino DM, Eaton S, Moss M. (2003) The epidemiology of sepsis in the United States from 1979 through 2000. N. Engl. J. Med. 348:1546–54.CrossRefGoogle Scholar
  3. 3.
    Deutschman CS, Tracey KJ. (2014) Sepsis: current dogma and new perspectives. Immunity. 40:463–75.CrossRefGoogle Scholar
  4. 4.
    Angus DC, van der Poll T. (2013) Severe sepsis and septic shock. N. Engl. J. Med. 369:840–51.CrossRefGoogle Scholar
  5. 5.
    Bone RC, et al. (2009) Definitions for sepsis and organ failure and guidelines for the use of innovative therapies in sepsis. The ACCP/SCCM Consensus Conference Committee. American College of Chest Physicians/Society of Critical Care Medicine. 1992. Chest. 136:e28.CrossRefGoogle Scholar
  6. 6.
    Aird WC. (2003) The role of the endothelium in severe sepsis and multiple organ dysfunction syndrome. Blood 101:3765–77.CrossRefGoogle Scholar
  7. 7.
    Aird WC. (2007) Endothelium as a therapeutic target in sepsis. Curr. Drug Targets. 8:501–7.CrossRefGoogle Scholar
  8. 8.
    Czabanka M, Peter C, Martin E, Walther A. (2007) Microcirculatory endothelial dysfunction during endotoxemia—insights into pathophysiology, pathologic mechanisms and clinical relevance. Curr. Vasc. Pharmacol. 5:266–75.CrossRefGoogle Scholar
  9. 9.
    Bockmeyer CL, et al. (2008) Inflammation-associated ADAMTS13 deficiency promotes formation of ultra-large von Willebrand factor. Haematol. 93:137–140.CrossRefGoogle Scholar
  10. 10.
    Kayal S, Jais JP, Aguini N, Chaudiere J, Labrousse J. (1998) Elevated circulating E-selectin, intercellular adhesion molecule 1, and von Willebrand factor in patients with severe infection. Am. J. Respir. Crit. Care Med. 157:776–84.CrossRefGoogle Scholar
  11. 11.
    Ruggeri ZM. (2003) von Willebrand factor, platelets and endothelial cell interactions. J. Thromb. Haemost. 1:1335–42.CrossRefGoogle Scholar
  12. 12.
    Arya M, et al. (2002) Ultralarge multimers of von Willebrand factor form spontaneous high-strength bonds with the platelet glycoprotein Ib-IX complex: studies using optical tweezers. Blood. 99:3971–7.CrossRefGoogle Scholar
  13. 13.
    Andrews RK, Berndt MC. (2004) Platelet physiology and thrombosis. Thromb. Res. 114:447–53.CrossRefGoogle Scholar
  14. 14.
    Uemura M, et al. (2005) Localization of ADAMTS13 to the stellate cells of human liver. Blood. 106:922–4.CrossRefGoogle Scholar
  15. 15.
    Levy GG, et al. (2001) Mutations in a member of the ADAMTS gene family cause thrombotic thrombocytopenic purpura. Nature. 413:488–94.CrossRefGoogle Scholar
  16. 16.
    Zhou Z, Nguyen TC, Guchhait P, Dong JF. (2010) von Willebrand factor, ADAMTS-13, and thrombotic thrombocytopenic purpura. Sem. Thromb. Hemost. 36:71–81.CrossRefGoogle Scholar
  17. 17.
    Symmers WS. (1952) Thrombotic microangiopathic haemolytic anaemia (thrombotic microangiopathy). Br. Med. J. 2:897–903.CrossRefGoogle Scholar
  18. 18.
    Wang Z, et al. (2010) Sepsis-induced disseminated intravascular coagulation with features of thrombotic thrombocytopenic purpura: a fatal fulminant syndrome. Clin. Appl. Thromb. Hemost. 17:251–3.CrossRefGoogle Scholar
  19. 19.
    Kinasewitz GT, Zein JG, Lee GL, Nazir SA, Taylor FB Jr (2005) Prognostic value of a simple evolving disseminated intravascular coagulation score in patients with severe sepsis. Crit. Care Med. 33:2214–21.CrossRefGoogle Scholar
  20. 20.
    Chertow GM, et al. (2006) Mortality after acute renal failure: models for prognostic stratification and risk adjustment. Kidney Int. 70:1120–6.CrossRefGoogle Scholar
  21. 21.
    Vincent JL, Yagushi A, Pradier O. (2002) Platelet function in sepsis. Crit. Care Med. 30:S313–7.CrossRefGoogle Scholar
  22. 22.
    Vischer UM. (2006) von Willebrand factor, endothelial dysfunction, and cardiovascular disease. J. Thromb. Haemost. 4:1186–93.CrossRefGoogle Scholar
  23. 23.
    Kremer Hovinga JA, et al. (2007) ADAMTS-13, von Willebrand factor and related parameters in severe sepsis and septic shock. J. Thromb. Haemost. 5:2284–90.CrossRefGoogle Scholar
  24. 24.
    Larkin D, et al. (2009) Severe Plasmodium falciparum malaria is associated with circulating ultra-large von Willebrand multimers and ADAMTS13 inhibition. PLoS Pathog. 5:e1000349.CrossRefGoogle Scholar
  25. 25.
    Peigne V, et al. (2013) The prognostic value of ADAMTS13 (a disintegrin and metalloprotease with thrombospondin type 1 repeats, member 13) deficiency in septic shock patients involves interleukin-6 and is not dependent on disseminated intravascular coagulation. Crit. Care. 17:R273.CrossRefGoogle Scholar
  26. 26.
    Claus RA, Bockmeyer CL, Sossdorf M, Losche W. (2010) The balance between von-Willebrand factor and its cleaving protease ADAMTS13: biomarker in systemic inflammation and development of organ failure? Curr. Mol. Med. 10:236–48.CrossRefGoogle Scholar
  27. 27.
    Nguyen TC, et al. (2008) Intensive plasma exchange increases ADAMTS-13 activity and reverses organ dysfunction in children with thrombocytopenia-associated multiple organ failure. Crit. Care Med. 36:2878–87.CrossRefGoogle Scholar
  28. 28.
    Bockmeyer CL, et al. (2011) ADAMTS13 activity is decreased in a septic porcine model. Significance for glomerular thrombus deposition. Thromb. Haemost. 105:145–53.CrossRefGoogle Scholar
  29. 29.
    Ono T, et al. (2006) Severe secondary deficiency of von Willebrand factor-cleaving protease (ADAMTS13) in patients with sepsis-induced disseminated intravascular coagulation: its correlation with development of renal failure. Blood. 107:528–34.CrossRefGoogle Scholar
  30. 30.
    Claus RA, et al. (2009) Variations in the ratio between von Willebrand factor and its cleaving protease during systemic inflammation and association with severity and prognosis of organ failure. Thromb. Haemost. 101:239–47.PubMedGoogle Scholar
  31. 31.
    Mannucci PM, et al. (2001) Changes in health and disease of the metalloprotease that cleaves von Willebrand factor. Blood. 98:2730–5.CrossRefGoogle Scholar
  32. 32.
    Cao WJ, Niiya M, Zheng XW, Shang DZ, Zheng XL. (2008) Inflammatory cytokines inhibit ADAMTS13 synthesis in hepatic stellate cells and endothelial cells. J. Thromb. Haemost. 6:1233–5.CrossRefGoogle Scholar
  33. 33.
    Bernardo A, Ball C, Nolasco L, Moake JF, Dong JF. (2004) Effects of inflammatory cytokines on the release and cleavage of the endothelial cell-derived ultralarge von Willebrand factor multimers under flow. Blood. 104:100–6.CrossRefGoogle Scholar
  34. 34.
    Crawley JT, et al. (2005) Proteolytic inactivation of ADAMTS13 by thrombin and plasmin. Blood. 105:1085–93.CrossRefGoogle Scholar
  35. 35.
    Mimuro J, et al. (2008) Unbalanced expression of ADAMTS13 and von Willebrand factor in mouse endotoxinemia. Thromb. Res. 122:91–7.CrossRefGoogle Scholar
  36. 36.
    Lerolle N, et al. (2009) von Willebrand factor is a major determinant of ADAMTS-13 decrease during mouse sepsis induced by cecum ligation and puncture. J. Thromb. Haemost. 7:843–50.CrossRefGoogle Scholar
  37. 37.
    Xu L, et al. (2005) Human hepatic stellate cell lines, LX-1 and LX-2: new tools for analysis of hepatic fibrosis. Gut. 54:142–51.CrossRefGoogle Scholar
  38. 38.
    Weiskirchen R, et al. (2013) Genetic characteristics of the human hepatic stellate cell line LX-2. PloS One. 8:e75692.CrossRefGoogle Scholar
  39. 39.
    Ades EW, et al. (1992) HMEC-1: establishment of an immortalized human microvascular endothelial cell line. J. Invest. Dermatol. 99:683–90.CrossRefGoogle Scholar
  40. 40.
    Ceppi ED, Smith FS, Titheradge MA. (1996) Effect of multiple cytokines plus bacterial endotoxin on glucose and nitric oxide production by cultured hepatocytes. Biochem. J. 317(Pt 2):503–7.CrossRefGoogle Scholar
  41. 41.
    Kuhn UD, Rost M, Muller D. (2001) Para-nitrophenol glucuronidation and sulfation in rat and human liver slices. Exp. Toxicol. Pathol. 53:81–7.CrossRefGoogle Scholar
  42. 42.
    Pfaffl MW. (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 29:e45.CrossRefGoogle Scholar
  43. 43.
    Kokame K, Nobe Y, Kokubo Y, Okayama A, Miyata T. (2005) FRETS-VWF73, a first fluorogenic substrate for ADAMTS13 assay. Br. J. Haematol. 129:93–100.CrossRefGoogle Scholar
  44. 44.
    Claus RA, Bockmeyer CL, Sossdorf M, Losche W, Hilberg T. (2006) Physical stress as a model to study variations in ADAMTS-13 activity, von Willebrand factor level and platelet activation. J. Thromb. Haemost. 4:902–5.CrossRefGoogle Scholar
  45. 45.
    Reuken PA, et al. (2014) Imbalance of von Willebrand factor and its cleaving protease ADAMTS13 during systemic inflammation superimposed on advanced cirrhosis. Liver Int. 35:37–45.CrossRefGoogle Scholar
  46. 46.
    Bai X, et al. (2014) Early versus delayed administration of norepinephrine in patients with septic shock. Crit. Care. 18:532.CrossRefGoogle Scholar
  47. 47.
    Pecori Giraldi F, et al. (2011) von Willebrand factor and fibrinolytic parameters during the desmopressin test in patients with Cushing’s disease. Br. J. Clin. Pharmacol. 71:132–6.CrossRefGoogle Scholar
  48. 48.
    Dziaman T, et al. (2009) Selenium supplementation reduced oxidative DNA damage in adnexectomized BRCA1 mutations carriers. Cancer Epidemiol. Biomarkers Prev. 18:2923–8.CrossRefGoogle Scholar
  49. 49.
    Christiaans SC, Wagener BM, Esmon CT, Pittet JF. (2013) Protein C and acute inflammation: a clinical and biological perspective. Am. J. Physiol. Lung Cell. Mol. Physiol. 305:L455–66.CrossRefGoogle Scholar
  50. 50.
    Turner NA, Nolasco L, Ruggeri ZM, Moake JL. (2009) Endothelial cell ADAMTS-13 and VWF: production, release, and VWF string cleavage. Blood. 114:5102–11.CrossRefGoogle Scholar
  51. 51.
    Kume Y, et al. (2007) Hepatic stellate cell damage may lead to decreased plasma ADAMTS13 activity in rats. FEBS Lett. 581:1631–4.CrossRefGoogle Scholar
  52. 52.
    Niiya M, et al. (2006) Increased ADAMTS-13 proteolytic activity in rat hepatic stellate cells upon activation in vitro and in vivo. J. Thromb. Haemost. 4:1063–70.CrossRefGoogle Scholar
  53. 53.
    Grillberger R, et al. (2014) A novel flow-based assay reveals discrepancies in ADAMTS-13 inhibitor assessment as compared with a conventional clinical static assay. J. Thromb. Haemost. 12:1523–32.CrossRefGoogle Scholar
  54. 54.
    Juul KV, Bichet DG, Nielsen S, Norgaard JP. (2014) The physiological and pathophysiological functions of renal and extrarenal vasopressin V2 receptors. Am. J. Physiol. renal Physiol. 306:F931–40.CrossRefGoogle Scholar
  55. 55.
    Reiter RA, Knobl P, Varadi K, Turecek PL. (2003) Changes in von Willebrand factor-cleaving protease (ADAMTS13) activity after infusion of desmopressin. Blood. 101:946–8.CrossRefGoogle Scholar
  56. 56.
    Reiter RA, Varadi K, Turecek PL, Jilma B, Knobl P. (2005) Changes in ADAMTS13 (von-Willebrand-factor-cleaving protease) activity after induced release of von Willebrand factor during acute systemic inflammation. Thromb. Haemost. 93:554–8.PubMedGoogle Scholar
  57. 57.
    Shomron N, et al. (2010) A splice variant of ADAMTS13 is expressed in human hepatic stellate cells and cancerous tissues. Thromb. Haemost 104:531–5.CrossRefGoogle Scholar
  58. 58.
    Alkozai EM, et al. (2015) No evidence for increased platelet activation in patients with hepatitis B- or C-related cirrhosis and hepatocellular carcinoma. Thromb. Res. 135:292–7.CrossRefGoogle Scholar
  59. 59.
    Fukushima H, et al. (2013) Ratio of von Willebrand factor propeptide to ADAMTS13 is associated with severity of sepsis. Shock. 39:409–14.CrossRefGoogle Scholar
  60. 60.
    Page AV, Liles WC. (2013) Biomarkers of endothelial activation/dysfunction in infectious diseases. Virulence. 4:507–16.CrossRefGoogle Scholar
  61. 61.
    Yamaguchi M, et al. (2006) Decreased protein C activation in patients with fulminant hepatic failure. Scand J. Gastroenterol. 41:331–7.CrossRefGoogle Scholar
  62. 62.
    Zhang F, et al. (2002) Inhibition of TNF-alpha induced ICAM-1, VCAM-1 and E-selectin expression by selenium. Atheroscler. 161:381–6.CrossRefGoogle Scholar
  63. 63.
    Tolando R, Jovanovic A, Brigelius-Flohe R, Ursini F, Maiorino M. (2000) Reactive oxygen species and proinflammatory cytokine signaling in endothelial cells: effect of selenium supplementation. Free Radic. Biol. Med. 28:979–86.CrossRefGoogle Scholar
  64. 64.
    Steinbrenner H, Bilgic E, Alili L, Sies H, Brenneisen P. (2006) Selenoprotein P protects endothelial cells from oxidative damage by stimulation of glutathione peroxidase expression and activity. Free Radic. Res. 40:936–43.CrossRefGoogle Scholar
  65. 65.
    Chauhan AK, et al. (2008) ADAMTS13: a new link between thrombosis and inflammation. J. Exp. Med. 205:2065–74.CrossRefGoogle Scholar
  66. 66.
    Heyland DK, Dhaliwal R, Suchner U, Berger MM. (2005) Antioxidant nutrients: a systematic review of trace elements and vitamins in the critically ill patient. Intensive Care Med. 31:327–37.CrossRefGoogle Scholar
  67. 67.
    Sakr Y, et al. (2014) Adjuvant selenium supplementation in the form of sodium selenite in postoperative critically ill patients with severe sepsis. Crit. Care. 18:R68.CrossRefGoogle Scholar
  68. 68.
    Forceville X. (2007) Effects of high doses of selenium, as sodium selenite, in septic shock patients a placebo-controlled, randomized, double-blind, multi-center phase II study—selenium and sepsis. J. Trace Elem. Med. Biol. 21 Suppl 1:62–5.CrossRefGoogle Scholar
  69. 69.
    Bosse AC, et al. (2010) Impact of selenite and selenate on differentially expressed genes in rat liver examined by microarray analysis. Biosci. Rep. 30:293–306.CrossRefGoogle Scholar
  70. 70.
    Dellinger RP, et al. (2013) Surviving sepsis campaign: international guidelines for management of severe sepsis and septic shock: 2012. Crit. Care Med. 41:580–637.CrossRefGoogle Scholar

Copyright information

© The Author(s) 2015

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, and provide a link to the Creative Commons license. You do not have permission under this license to share adapted material derived from this article or parts of it.

The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.

To view a copy of this license, visit (

Authors and Affiliations

  • Michael L. Ekaney
    • 1
    • 2
  • Clemens L. Bockmeyer
    • 3
  • Maik Sossdorf
    • 1
    • 2
  • Philipp A. Reuken
    • 1
    • 2
  • Florian Conradi
    • 2
  • Tobias Schuerholz
    • 4
  • Markus F. Blaess
    • 1
    • 2
  • Scott L. Friedman
    • 5
  • Wolfgang Lösche
    • 1
    • 2
  • Michael Bauer
    • 1
    • 2
  • Ralf A. Claus
    • 1
    • 2
  1. 1.Center for Sepsis Control and Care, and Clinic for Anaesthesiology and Intensive Care TherapyJena University HospitalJenaGermany
  2. 2.Clinic for Anaesthesiology and Intensive Care MedicineJena University HospitalJenaGermany
  3. 3.Department of NephropathologyUniversity Hospital AachenAachenGermany
  4. 4.Department for Interdisciplinary Intensive CareUniversity Hospital AachenAachenGermany
  5. 5.Mount Sinai School of MedicineNew YorkUSA

Personalised recommendations