Download PDF
Thromb Haemost 2017; 117(03): 570-579
DOI: 10.1160/TH16-10-0762
Blood Cells, Inflammation and Infection
Schattauer GmbH

The fifth epidermal growth factor like region of thrombomodulin alleviates LPS-induced sepsis through interacting with GPR15

Bin Pan
1   Department of Hematology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, Jiangsu, China
2   Department of Hematology and Respiratory Medicine, Kochi Medical School, Kochi University, Nankoku, Kochi, Japan
3   Department of Hematology, Fukushima Medical University, Fukushima, Japan
,
Xiangmin Wang
1   Department of Hematology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, Jiangsu, China
2   Department of Hematology and Respiratory Medicine, Kochi Medical School, Kochi University, Nankoku, Kochi, Japan
3   Department of Hematology, Fukushima Medical University, Fukushima, Japan
,
Shinsuke Kojima
2   Department of Hematology and Respiratory Medicine, Kochi Medical School, Kochi University, Nankoku, Kochi, Japan
,
Chie Nishioka
2   Department of Hematology and Respiratory Medicine, Kochi Medical School, Kochi University, Nankoku, Kochi, Japan
4   Department of Immunology, Kochi Medical School, Kochi University, Nankoku, Kochi, Japan
,
Akihito Yokoyama
2   Department of Hematology and Respiratory Medicine, Kochi Medical School, Kochi University, Nankoku, Kochi, Japan
,
Goichi Honda
5   Medical Affairs Department, Asahi Kasei Pharma., Kanda Jinbocho, Chiyoda-ku, Tokyo, Japan
,
Kailin Xu
1   Department of Hematology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, Jiangsu, China
,
Takayuki Ikezoe
2   Department of Hematology and Respiratory Medicine, Kochi Medical School, Kochi University, Nankoku, Kochi, Japan
3   Department of Hematology, Fukushima Medical University, Fukushima, Japan
› Author Affiliations Financial support: This study was supported in part by SENSHIN Medical Research Foundation (T.I), and KAKENHI (26461406) (T.I).
Further Information

Publication History

Received: 07 October 2016

Accepted after major revision: 02 January 2016

Publication Date:
22 November 2017 (online)

    Permissions and Reprints

Summary

Thrombomodulin (TM) exerts cytoprotection via the fifth region of epidermal growth factor (EGF)-like domain of TM (TME5) by interacting with G-protein coupled receptor 15 (GPR15) expressed on cell surface of vascular endothelial cells. TM is also implied to mediate anti-inflammatory functions by unknown mechanism. By applying a lipopolysaccharide (LPS)-induced murine sepsis model, we assessed the role of TME5 in septic inflammation and coagulation. We found that TME5 treatment protected mice in association with ameliorating inflammation and coagulopathy in LPS-induced sepsis. Further study confirmed that TME5 bound GPR15 in vitro. Knock out of GPR15 abolished protective role of TME5 in sepsis model. GPR15 mediated anti-inflammatory function of TME5 through suppression of phosphorylation of IκBα, nuclear translocation of NF-κB and release of pro-inflammatory cytokines in macro-phages (Macs). Knock out of GPR15 resulted in dysregulated immune response of Macs, characterised by excessive expression of pro-inflammatory genes and failing to limit immune response. This study indicates that TME5 exerts anti-inflammatory function through inhibition of NF-κB in a GPR15-dependent manner. The use of TME5 may be a potential therapeutic option for treatment of sepsis.

Supplementary Material to this article is available online at www.thrombosis-online.com.

Keywords

Thrombomodulin - lipopolysaccharide - sepsis - G-protein coupled receptor 15 - macrophages

 

  • References

  • 1 Iwasaki A, Medzhitov R. Control of adaptive immunity by the innate immune system. Nat Immunol 2015; 16: 343-353.
    CrossrefPubMedGoogle Scholar
  • 2 Hotchkiss RS, Monneret G, Payen D. Sepsis-induced immunosuppression: from cellular dysfunctions to immunotherapy. Nat Rev Immunol 2013; 13: 862-874.
    CrossrefPubMedGoogle Scholar
  • 3 Remick DG. Cytokine therapeutics for the treatment of sepsis: why has nothing worked?. Curr Pharm Des 2003; 09: 75-82.
    CrossrefPubMedGoogle Scholar
  • 4 Maugeri N, Brambilla M, Camera M. et al. Human polymorphonuclear leukocytes produce and express functional tissue factor upon stimulation. J Thromb Haemost 2006; 04: 1323-1330.
    CrossrefPubMedGoogle Scholar
  • 5 Opal SM, van der Poll T. Endothelial barrier dysfunction in septic shock. J Intern Med 2015; 277: 277-293.
    CrossrefPubMedGoogle Scholar
  • 6 Suzuki K, Kusumoto H, Deyashiki Y. et al. Structure and expression of human thrombomodulin, a thrombin receptor on endothelium acting as a cofactor for protein C activation. EMBO J 1987; 06: 1891-1897.
    PubMedGoogle Scholar
  • 7 Conway EM, Van de Wouwer M, Pollefeyt S. et al. The lectin-like domain of thrombomodulin confers protection from neutrophil-mediated tissue damage by suppressing adhesion molecule expression via nuclear factor kappaB and mitogen-activated protein kinase pathways. J Exp Med 2002; 196: 565-577.
    CrossrefPubMedGoogle Scholar
  • 8 Cao C, Gao Y, Li Y. et al. The efficacy of activated protein C in murine endotoxemia is dependent on integrin CD11b. J Clin Invest 2010; 120: 1971-1980.
    CrossrefPubMedGoogle Scholar
  • 9 Dittman WA, Majerus PW. Structure and function of thrombomodulin: a natural anticoagulant. Blood 1990; 75: 329-336.
    PubMedGoogle Scholar
  • 10 Isermann B, Vinnikov IA, Madhusudhan T. et al. Activated protein C protects against diabetic nephropathy by inhibiting endothelial and podocyte apoptosis. Nat Med 2007; 13: 1349-1358.
    CrossrefPubMedGoogle Scholar
  • 11 Ito T, Kakihana Y, Maruyama I. Thrombomodulin as an intravascular safeguard against inflammatory and thrombotic diseases. Expert Opin Ther Targets 2016; 20: 151-158.
    CrossrefPubMedGoogle Scholar
  • 12 Saito H, Maruyama I, Shimazaki S. et al. Efficacy and safety of recombinant human soluble thrombomodulin (ART-123) in disseminated intravascular coagulation: results of a phase III, randomized, double-blind clinical trial. J Thromb Haemost 2007; 05: 31-41.
    CrossrefPubMedGoogle Scholar
  • 13 Sakai M, Ikezoe T, Bandobashi K. et al. Successful treatment of transplantation-associated thrombotic microangiopathy with recombinant human soluble thrombomodulin. Bone Marrow Transplant 2010; 45: 803-805.
    CrossrefPubMedGoogle Scholar
  • 14 Ikezoe T, Togitani K, Komatsu N. et al. Successful treatment of sinusoidal obstructive syndrome after hematopoietic stem cell transplantation with recombinant human soluble thrombomodulin. Bone Marrow Transplant 2010; 45: 783-785.
    CrossrefPubMedGoogle Scholar
  • 15 Ikezoe T, Takeuchi A, Taniguchi A. et al. Recombinant human soluble thrombomodulin counteracts capillary leakage associated with engraftment syndrome. Bone Marrow Transplant 2011; 46: 616-618.
    CrossrefPubMedGoogle Scholar
  • 16 Ikezoe T, Yang J, Nishioka C. et al. Thrombomodulin protects endothelial cells from a calcineurin inhibitor-induced cytotoxicity by upregulation of extracellular signal-regulated kinase/myeloid leukemia cell-1 signalling. Arterioscler Thromb Vasc Biol 2012; 32: 2259-2270.
    CrossrefPubMedGoogle Scholar
  • 17 Ikezoe T, Yang J, Nishioka C. et al. The fifth epidermal growth factor-like region of thrombomodulin exerts cytoprotective function and prevents SOS in a murine model. Bone Marrow Transplant. 2016 Epub ahead of print.
    PubMedGoogle Scholar
  • 18 Cartwright A, Schmutz C, Askari A. et al. Orphan receptor GPR15/BOB is up-regulated in rheumatoid arthritis. Cytokine 2014; 67: 53-59.
    CrossrefPubMedGoogle Scholar
  • 19 Koks G, Uudelepp ML, Limbach M. et al. Smoking-induced expression of the GPR15 gene indicates its potential role in chronic inflammatory pathologies. Am J Pathol 2015; 185: 2898-2906.
    CrossrefPubMedGoogle Scholar
  • 20 Nguyen LP, Pan J, Dinh TT. et al. Role and species-specific expression of colon T cell homing receptor GPR15 in colitis. Nat Immunol 2015; 16: 207-213.
    CrossrefPubMedGoogle Scholar
  • 21 Kim SV, Xiang WV, Kwak C. et al. GPR15-mediated homing controls immune homeostasis in the large intestine mucosa. Science 2013; 340: 1456-1459.
    CrossrefPubMedGoogle Scholar
  • 22 Pan B, Liu J, Zhang Y. et al. Acute ablation of DP thymocytes induces up-regulation of IL-22 and Foxn1 in TECs. Clin Immunol 2014; 150: 101-108.
    CrossrefPubMedGoogle Scholar
  • 23 Lappin E, Ferguson AJ. Gram-positive toxic shock syndromes. Lancet Infect Dis 2009; 09: 281-290.
    CrossrefPubMedGoogle Scholar
  • 24 Chen Z, Ding X, Jin S. et al. WISP1-alphavbeta3 integrin signalling positively regulates TLR-triggered inflammation response in sepsis induced lung injury. Sci Rep 2016; 06: 28841.
    CrossrefPubMedGoogle Scholar
  • 25 Packiriswamy N, Parameswaran N. G-protein-coupled receptor kinases in inflammation and disease. Genes Immun 2015; 16: 367-377.
    CrossrefPubMedGoogle Scholar
  • 26 Rittirsch D, Flierl MA, Ward PA. Harmful molecular mechanisms in sepsis. Nat Rev Immunol 2008; 08: 776-787.
    CrossrefPubMedGoogle Scholar
  • 27 Blann AD, Woywodt A, Bertolini F. et al. Circulating endothelial cells. Biomarker of vascular disease. Thromb Haemost 2005; 93: 228-235.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 28 Esmon CT. Thrombomodulin as a model of molecular mechanisms that modulate protease specificity and function at the vessel surface. FASEB J 1995; 09: 946-955.
    CrossrefPubMedGoogle Scholar
  • 29 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.
    CrossrefPubMedGoogle Scholar
  • 30 Ito T, Maruyama I. Thrombomodulin: protectorate God of the vasculature in thrombosis and inflammation. J Thromb Haemost 2011; 09 (Suppl. 01) 168-173.
    CrossrefPubMedGoogle Scholar
  • 31 Shi CS, Shi GY, Hsiao HM. et al. Lectin-like domain of thrombomodulin binds to its specific ligand Lewis Y antigen and neutralizes lipopolysaccharide-induced inflammatory response. Blood 2008; 112: 3661-3670.
    CrossrefPubMedGoogle Scholar
  • 32 Yin Q, Fu TM, Li J. et al. Structural biology of innate immunity. Annu Rev Immunol 2015; 33: 393-416.
    CrossrefPubMedGoogle Scholar
  • 33 Korn T, Bettelli E, Oukka M. et al. IL-17 and Th17 Cells. Annu Rev Immunol 2009; 27: 485-517.
    CrossrefPubMedGoogle Scholar
  • 34 Zhou L, Chong MM, Littman DR. Plasticity of CD4+ T cell lineage differentiation. Immunity 2009; 30: 646-655.
    CrossrefPubMedGoogle Scholar
  • 35 Deng HK, Unutmaz D, KewalRamani VN. et al. Expression cloning of new receptors used by simian and human immunodeficiency viruses. Nature 1997; 388: 296-300.
    CrossrefPubMedGoogle Scholar
  • 36 Lahl K, Sweere J, Pan J. et al. Orphan chemoattractant receptor GPR15 mediates dendritic epidermal T-cell recruitment to the skin. Eur J Immunol 2014; 44: 2577-2581.
    CrossrefPubMedGoogle Scholar
  • 37 Hirayama F, Ogawa M. Cytokine regulation of early lymphohematopoietic development. Stem Cells 1996; 14: 369-375.
    CrossrefPubMedGoogle Scholar
  • 38 Shakoory B, Fitzgerald SM, Lee SA. et al. The role of human mast cell-derived cytokines in eosinophil biology. J Interferon Cytokine Res 2004; 24: 271-281.
    CrossrefPubMedGoogle Scholar
  • 39 Villarino AV, Kanno Y, Ferdinand JR. et al. Mechanisms of Jak/STAT signalling in immunity and disease. J Immunol 2015; 194: 21-27.
    CrossrefPubMedGoogle Scholar
  • 40 Yoshie O, Matsushima K. CCR4 and its ligands: from bench to bedside. Int Immunol 2015; 27: 11-20.
    CrossrefPubMedGoogle Scholar
  • 41 Dejda A, Mawambo G, Cerani A. et al. Neuropilin-1 mediates myeloid cell chemoattraction and influences retinal neuroimmune crosstalk. J Clin Invest 2014; 124: 4807-4822.
    CrossrefPubMedGoogle Scholar
  • 42 Vorup-Jensen T, Ostermeier C, Shimaoka M. et al. Structure and allosteric regulation of the alpha X beta 2 integrin I domain. Proc Natl Acad Sci USA 2003; 100: 1873-1878.
    CrossrefPubMedGoogle Scholar
  • 43 Ruiz S, Santos E, Bustelo XR. RasGRF2, a guanosine nucleotide exchange factor for Ras GTPases, participates in T-cell signalling responses. Mol Cell Biol 2007; 27: 8127-8142.
    CrossrefPubMedGoogle Scholar
  • 44 Sica A, Mantovani A. Macrophage plasticity and polarization: in vivo veritas. J Clin Invest 2012; 122: 787-795.
    CrossrefPubMedGoogle Scholar