Increased levels of citrullinated antithrombin in plasma of patients with rheumatoid arthritis and colorectal adenocarcinoma determined by a newly developed ELISA using a specific monoclonal antibody

Adriana Ordóñez; Jose Yélamos; Shona Pedersen; Antonia Miñano; Pablo Conesa-Zamora; Soren R. Kristensen; Mogens T. Stender; Ole Thorlacius-Ussing; Irene Martínez-Martínez; Vicente Vicente; Javier Corral

doi:10.1160/TH10-05-0297

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2010; 104(06): 1143-1149
DOI: 10.1160/TH10-05-0297

Blood Coagulation, Fibrinolysis and Cellular Haemostasis

Schattauer GmbH

Increased levels of citrullinated antithrombin in plasma of patients with rheumatoid arthritis and colorectal adenocarcinoma determined by a newly developed ELISA using a specific monoclonal antibody

Adriana Ordóñez

¹Centro Regional de Hemodonación, University of Murcia, Spain

,

Jose Yélamos

²Department of Immunology, Cancer Program, IMIM-Hospital del Mar, Barcelona, Spain

,

Shona Pedersen

³Aarhus University Hospital, Aalborg Hospital, Clinical Biochemistry, Centre for Cardiovascular Research, Aalborg, Denmark

,

Antonia Miñano

¹Centro Regional de Hemodonación, University of Murcia, Spain

,

Pablo Conesa-Zamora

⁴Grupo de Patología Molecular y Farmacogenética, Hospital Universitario Santa María del Rosell, Cartagena, Spain

,

Soren R. Kristensen

³Aarhus University Hospital, Aalborg Hospital, Clinical Biochemistry, Centre for Cardiovascular Research, Aalborg, Denmark

,

Mogens T. Stender

⁵Aalborg Hospital, Aarhus University Hospital, Department of Surgical Gastroenterology, Aalborg, Jutland, Denmark

,

Ole Thorlacius-Ussing

⁵Aalborg Hospital, Aarhus University Hospital, Department of Surgical Gastroenterology, Aalborg, Jutland, Denmark

,

Irene Martínez-Martínez

¹Centro Regional de Hemodonación, University of Murcia, Spain

,

Vicente Vicente

¹Centro Regional de Hemodonación, University of Murcia, Spain

,

Javier Corral

¹Centro Regional de Hemodonación, University of Murcia, Spain

› Institutsangaben

Abstract

Summary

Citrullination is a post-translational modification that plays essential roles in both physiological processes and disease. Recent studies have found increased levels of citrullinated antithrombin in patients with rheumatoid arthritis and in different malignant tumours. Antithrombin, the main haemostatic serpin, loses its anticoagulant function via citrullination, which might contribute to the pathogenesis or thrombotic side effects of these disorders. We have developed a specific monoclonal antibody against citrullinated antithrombin. We determined the levels of citrullinated antithrombin and anti-FXa activity in plasma from 66 donors, 17 patients with rheumatoid arthritis and 77 patients with colorectal adenocarcinoma (42 suffering from venous thrombosis). Healthy subjects had negligible amounts of citrullinated antithrombin in plasma (7.9 ± 22.1 ng/ml), while it significantly increased in patients with rheumatoid arthritis or adenocarcinoma (159.7 ± 237.6 ng/ml and 36.8 ± 66.1 ng/ml), levels that, however, did not modify the plasma anticoagulant activity. Moreover, we did not find association between citrullinated antithrombin and the thrombotic risk in patients with adenocarcinoma. In conclusion, we have developed an antibody specific for citrullinated antithrombin that allows its quantification in biological samples, offering a new tool for the analysis of citrullination in different diseases. We confirm increased levels of citrullinated antithrombin in plasma of patients with rheumatoid arthritis and adenocarcinoma. This modification, probably local, could have pathological consequences in both disorders, but only affects a minor fraction of plasma antithrombin, resulting in no significant reduction of global anticoagulant activity. This result explains the absence of association of this marker with an increased risk of thrombosis in patients with colorectal adenocarcinoma.

Keywords

Antithrombin - citrullination - monoclonal antibody - rheumatoid arthritis - cancer

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Referenzen

References
1 Kouzarides T. Acetylation: a regulatory modification to rival phosphorylation?. EMBO J 2000; 19: 1176-1179.
2 Wells L. et al. Glycosylation of nucleocytoplasmic proteins: signal transduction and O-GlcNAc. Science 2001; 291: 2376-2378.
3 Akimoto Y. et al. Elevation of the post-translational modification of proteins by O-linked N-acetylglucosamine leads to deterioration of the glucose-stimulated insulin secretion in the pancreas of diabetic Goto-Kakizaki rats. Glycobiology 2007; 17: 127-140.
4 Kim JK. et al. Multiple sclerosis: an important role for post-translational modifications of myelin basic protein in pathogenesis. Mol Cell Proteomics 2003; 2: 453-462.
5 Ishida-Yamamoto A. et al. Decreased deiminated keratin K1 in psoriatic hyper-proliferative epidermis. J Invest Dermatol 2000; 114: 701-705.
6 Moscarello MA. et al. The role of citrullinated proteins suggests a novel mechanism in the pathogenesis of multiple sclerosis. Neurochem Res 2007; 32: 251-256.
7 Nijenhuis S. et al. Autoantibodies to citrullinated proteins in rheumatoid arthritis: clinical performance and biochemical aspects of an RA-specific marker. Clin Chim Acta 2004; 350: 17-34.
8 Suzuki A. et al. Citrullination by peptidylarginine deiminase in rheumatoid arthritis. Ann NY Acad Sci 2007; 1108: 323-339.
9 Vossenaar ER. et al. PAD, a growing family of citrullinating enzymes: genes, features and involvement in disease. Bioessays 2003; 25: 1106-1118.
10 van Venrooij WJ, Pruijn GJ. Citrullination: A small change for a protein with great consequences for rheumatoid arthritis. Arthritis Res 2000; 2: 249-251.
11 Tarcsa E. et al. Protein unfolding by peptidylarginine deiminase. Substrate specificity and structural relationships of the natural substrates trichohyalin and filaggrin. J Biol Chem 1996; 271: 30709-30716.
12 Gyorgy B. et al. Citrullination: a posttranslational modification in health and disease. Int J Biochem Cell Bio 2006; 38: 1662-1677.
13 Asaga H. et al. Selective deimination of vimentin in calcium ionophore-induced apoptosis of mouse peritoneal macrophages. Biochem Biophys Res Commun 1998; 243: 641-646.
14 Jang B. et al. Involvement of peptidylarginine deiminase-mediated post-translational citrullination in pathogenesis of sporadic Creutzfeldt-Jakob disease. Acta Neuropathol 2010; 119: 199-210.
15 Tanikawa C. et al. Regulation of protein Citrullination through p53/PADI4 network in DNA damage response. Cancer Res 2009; 69: 8761-8769.
16 Loos T. et al. Citrullination of CXCL10 and CXCL11 by peptidylarginine deiminase: a naturally occurring posttranslational modification of chemokines and new dimension of immunoregulation. Blood 2008; 112: 2648-2656.
17 Hernández-Espinosa D. et al. Factors with conformational effects on haemostatic serpins: implications in thrombosis. Thromb Haemost 2007; 98: 557-563.
18 Chang X. et al. The inhibition of antithrombin by peptidylarginine deiminase 4 may contribute to pathogenesis of rheumatoid arthritis. Rheumatology (Oxford) 2005; 44: 293-298.
19 Pike RN. et al. Heparin-dependent modification of the reactive center arginine of antithrombin and consequent increase in heparin binding affinity. J Biol Chem 1997; 272: 19652-19655.
20 Ordóñez A. et al. Effect of citrullination on the function and conformation of antithrombin. FEBS J 2009; 276: 6763-6772.
21 Ishiguro K. et al. Complete antithrombin deficiency in mice results in embryonic lethality. J Clin Invest 2000; 106: 873-878.
22 Chang X. et al. Increased PADI4 expression in blood and tissues of patients with malignant tumors. BMC Cancer 2009; 9: 40.
23 Sud R, Khorana AA. Cancer-associated thrombosis: risk factors, candidate biomarkers and a risk model. Thromb Res 2009; 123 Suppl S18-21.
24 Kuderer NM. et al. Impact of venous thromboembolism and anticoagulation on cancer and cancer survival. Clin Oncol 2009; 27: 4902-4911.
25 Mameli A. et al. Rheumatoid arthritis and thrombosis. Clin Exp Rheumatol 2009; 27: 846-855.
26 Matta F. et al. Risk of venous thromboembolism with rheumatoid arthritis. Thromb Haemost 2009; 101: 134-138.
27 Köhler G, Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 1975; 256: 495-497.
28 Stender MT. et al. Preoperative plasma D-dimer is a predictor of postoperative deep venous thrombosis in colorectal cancer patients: a clinical, prospective cohort study with one-year follow-up. Dis Colon Rectum 2009; 52: 446-451.
29 Wegner N. et al. Autoimmunity to specific citrullinated proteins gives the first clues to the etiology of rheumatoid arthritis. Immunol Rev 2010; 233: 34-54.
30 Yamada R. et al. Citrullinated proteins in rheumatoid arthritis. Front Biosci 2005; 10: 54-64.
31 Schellekens GA. et al. The diagnostic properties of rheumatoid arthritis antibodies recognizing a cyclic citrullinated peptide. Arthritis Rheum 2000; 43: 155-163.
32 Pruijn GJ. et al. The use of citrullinated peptides and proteins for the diagnosis of rheumatoid arthritis. Arthritis Res Ther 2010; 12: 203.
33 van Venrooij WJ. et al. Anti-CCP Antibody, a Marker for the Early Detection of Rheumatoid Arthritis. Ann NY Acad Sci 2008; 1143: 268-285.
34 van Gaalen FA. et al. Autoantibodies to cyclic citrullinated peptides predict progression to rheumatoid arthritis in patients with undifferentiated arthritis: a prospective cohort study. Arthritis Rheum 2004; 50: 709-715.
35 Szekanecz Z. et al. Anti-citrullinated protein antibodies in rheumatoid arthritis: as good as it gets?. Clin Rev Allergy Immunol 2008; 34: 26-31.
36 Gabriel SE. Cardiovascular morbidity and mortality in rheumatoid arthritis. Am J Med 2008; 121: S9-14.
37 Undas A. et al. Thrombin generation in rheumatoid arthritis: dependence on plasma factor composition. Thromb Haemost 2010; 104: 224-230.
38 Chang X, Han J. Expression of peptidylarginine deiminase type 4 (PAD4) in various tumors. Mol Carcinog 2006; 45: 183-196.
39 Cohen AT. et al. Assessment of venous thromboembolism risk and the benefits of thromboprophylaxis in medical patients. Thromb Haemost. 2005; 94: 750-759.
40 Boccaccio C, Comoglio PMJ. Genetic link between cancer and thrombosis. Clin Oncol 2009; 27: 4827-4833.
41 Khong TL. et al. Angiogenesis as a therapeutic target in arthritis: learning the lessons of the colorectal cancer experience. Angiogenesis 2007; 10: 243-258.
42 O’Reilly MS. Antiangiogenic antithrombin. Semin Thromb Hemost 2007; 33: 660-666.