EXTRACELLULAR DNA AND THE LEVEL OF ITS METHYLATION IN DIFFERENT RHEUMATIC DISEASES

Objective: to study the qualitative and quantitative composition of serum extracellular DNA (exDNA); apoptotic (nucleosomes) and necrotic fragmentation; complexification of monometinic proteins with exDNA; and the levels of exDNA methylation as compared to those of autoimmune and inflammatory markers in patients with rheumatic diseases (RD). Subjects and methods. One hundred and one patients, including 28 with rheumatoid arthritis (RA), 20 with systemic lupus erythematosus (SLE), 14 with ankylosing spondylitis (AS), 31 with scleroderma systematica (SDS), and 8 with other RDs, were examined. All the patients met the classification criteria accepted at the Research Institute of Rheumatology. A control group included 15 healthy individuals. The investigators studied the serum concentration of exDNA, the content of DNA fragments of different lengths and the monometinic protein complexified with DNA, the degree of DNA methylation, the levels of serum C-reactive protein (CRP) detectable by a high-sensitivity method, rheumatoid factor (RF), anti-DNA antibodies, and antinuclear antibodies (ANA).
Results. 50-70% of the patients with RA, SLE, AS, and SDS were found to have elevated serum levels of exDNA, apoptotic and necrotic DNA; the degree of exDNA methylation (hypo- and hypermethylation) was altered. In SLE and SDS, exDNA hypomethylation was attended by autoimmune disorders: the presence of proper DNA autoantibodies and ANA. In RA, the high levels of exDNA methylation were associated with autoimmunity and inflammation (increased RF and CRP levels). In AS, the hypermethylation of apoptotic and necrotic DNA and monometinic proteins correlated with inflammatory activity and autoantigenicity (the appearance of ANA and anti-neutrophil antibodies).
Conclusion. RDs are characterized by the higher concentration of apoptotic and necrotic DNA, impaired exDNA methylation, varying complexification of exDNA with monometinic proteins, which is associated with the hyperproduction of autoantibodies (including anti-exDNA antibodies) and inflammatory markers.

extracellular DNA, methylation, apoptotic and necrotic DNA, autoantibodies, nucleosomes, rheumatic diseases

1. <div><p>Ревматология. Клинические рекомендации. Под ред.</p><p>Е.Л. Насонова. М.: ГЭОТАР-Медиа, 2008.</p><p>Насонов Е.Л. Перспективы развития ревматологии в XXI веке. Рус мед журн 2001;9:1031 -7.</p><p>Siegal L.M., Lypsky P. Autoimmunity. In: F. Kelly. Textbook of Rheumatology. Philadelphia: W.B. Saunders Company, 2008.</p><p>Насонов Е.Л., Соловьев С.К., Ананьева Л.П. и др. Ритуксимаб: прогресс в лечении аутоиммунных ревматических болезней. Науч-практич ревматол 2010;4:3-9.</p><p>Насонова В.А. Ревматология: взгляд в XXI век. Вестн РАМН 2003;71:3-6.</p><p>Насонова В.А. Системная красная волчанка. В кн.: Ревматизм и другие коллагеновые болезни. Методические указания МЗ СССР. М., 1961;108 с.</p><p>Guendel I., Carpio L., Pedati C. et al. Methylation of the tumor suppressor protein, BRCA1, influences its transcriptional cofactor function. PLoS One 2010;29:11379.</p><p>Гвоздев В.А. Регуляция активности генов, обусловленная химической модификацией (метилированием) ДНК. Соросовский образоват журн 1999;10:11-7.</p><p>Александрова Е.Н., Шубаева Н.О., Купавцева О.А. и др. Метилирование экстрацеллюлярной ДНК при различных ревматических заболеваниях: Доклад. В кн.: VIII конференция ревматологов Урала, 28-30 июня 2010 г.</p><p>Steinman C.R. Circulating DNA in systemic lupus erythematosus. Isolation and characterization. J Clin Invest 1984;73(3):832-41.</p><p>Mortensen E.S., Rekvig O.P. Nephritogenic potential of anti-DNA antibodies against necrotic nucleosomes. J Am Soc Nephrol Review 2009;20:696-704.</p><p>Seredkina N., Zykova S.N., Rekvig O.P. Progression of murine lupus nephritis is linked to acquired renal Dnase1 deficiency and not to up-regulated apoptosis. Am J Pathol 2009;175:97-106.</p><p>Urbonaviciute V., Fürnrohr B.G., Meister S. et al. Induction of inflammatory and immune responses by HMGB1-nucleosome complexes: implications for the pathogenesis of SLE. J Exp Med 2008;205:3007-18.</p><p>Robertson K.D. DNA methylation and human disease. Nature Rev Genetics 2005;6:597-610.</p><p>Laird P. The power and the promise of DNA methylation markers. Nature Reviews Cancer 2003;3:253-66.</p><p>Вейко Н.Н. Структурно-функциональная организация рибосомных повторов человека: Дис. ... д-ра биол наук. М., 2001;121 с.</p><p>Sambrook J., Fritsch E.F., Maniatis T. Molecular cloning: a laboratory manual. 2nd ed. NY: Cold Spring Harbor Laboratory Press, Cold Spring Harbor, 1989.</p><p>Wahlfors J., Hiltunen H., Heinonen K. et al. Genomic hypomethylation in human chronic lymphocytic leukemia. Blood 1992;80:2074-80.</p><p>Cote B.D., Uhlenbeck O.C., Steffensen D.M. Quantitation of in situ hybridization of ribosomal ribonucleic acids to human diploid cells. Cromosoma 1980;80:349-67.</p><p>Давыдовский И.В. К вопросу об острой красной волчанке. Рус вестн дерматол 1929;7:450-72.</p><p>Gopalakrishnan S., van Emburgh B.O., Robertson D. DNA metylation in development and human disease. Mutat Res 2008;647(1-2):30-8.</p><p>Areti R., Polidoros K., Panayiotis M. et al. Systemic lupus erythematosus and epigenetics. E-Journal Sci Technol(e-JST) 2009;4:75-85.</p><p>Richardson B., Scheinbart L., Strahler J. et al. Evidence for impaired T cell DNA methylation in systemic lupus erythematosus and rheumatoid arthritis. Arthr Rheum 1990;33:1665-72.</p><p>Yung R.L., Richardson B.C. Role of T-cells DNA Methylation in lupus syndromes. Lupus 1994;3:487-91.</p><p>Bruns A., Bläss S., Hausdorf G. et al. Nucleosomes are major T and B cell autoantigens in systemic lupus erythematosus. Arthr Rheum 2000;43:2307-15.</p><p>Denfeld R.W., Kind P., Sontheimer R.D. et al. In situ expression of B7 and CD28 receptor families in skin lesions of patients with lupus erythematosus. Arthr Rheum 1997;40:814-21.</p><p>Науч-практич ревматол 2012; 50(1): 22-27</p><p>Mohan C., Adams S., Stanik V. et al. Nucleosome: a major immunogen for pathogenic autoantibody-inducing T cells of lupus. J Exs Med 1993;177:1367-81.</p><p>Rekvig O.P., Moens U., Sudsjord A. et al. Experimental expression in mice and spontaneous expression in human SLE of polyomavirus E-antigen. A molecular basis for induction of antibodies to DNA and eukaryotic cells. J Clin Invest 1997;99:2045-54.</p><p>Fleischhaker M., Schidt B. Circulating nucleic acids (CNAs). Biochim Biophys Acta 2007;1775:181-232.</p><p>Kalaaji M., Mortensen E., Jorgensen L. et al. Nephritogenic lupus antibodies recognize glomerular basement membrane-associated chromatin fragments released from apoptotic intraglomerular cells. Am J Pathol 2006;168:1779-92.</p><p>Takami N., Osawa K., Miura Y. et.al. Hypermethylated promoter region of DR3, the death receptor 3 gene, in rheumatoid arthritis synovial cells. Arthr Rheum 2006;54:779-87.</p><p>Henault J., Robitaille G., Senecal J.L. et al. DNA topoisomerase I binding to fibroblasts induces monocyte adhesion and activation in the presence of anti-topoisomerase I autoantibodies from systemic sclerosis patients. Arthr Rheum 2006;54:963-73.</p><p>Hayakawa I., Hasegawa M., Takehara K., Sato S. Anti-DNA topoisomerase II autoantibodies in localized scleroderma. Arthr Rheum 2004;50:227-32.</p><p>Poscl A., Fidler A., Scmidt B. et al. DNA methylation is not likely to be responsible for aggrecan down regulation in aged or osteoarthtitic cartilage. Ann Rheum Dis 2005;64:477-80.</p><p>Gugliesi F., De Andrea M., Mondini M. et al. The proapoptotic activity of the Interferoh-inducible gene IFI16 provides new insights into its etiopathogenetic role in autoimmunity. J Autoimmun 2010;31:114-23.</p></div><br />