MOLECULAR GENETIC TESTING OF ACCP-POSITIVE PATIENTS WITH RHEUMATOID ARTHRITIS AND HIGH INFLAMMATORY DISEASE ACTIVITY (A REMARCA STUDY)

Rheumatoid arthritis (RA) is a multifactorial disease, in which the interaction of the genetic component and environmental factors, determines not only the development of the disease, but also its pronounced clinical polymorphism. We assume that the high inflammatory activity of RA may be determined by the genes, the products of which trigger inflammatory processes.

Objective: to investigate allele and genotype distribution of gene polymorphic variants in active anti-cyclic citrullinated peptide (aCCP)-positive patients with RA from the REMARCA program versus a control group of healthy blood donors.

Subjects and methods. A molecular genetic study enrolled 146 aCCP-positive patients from the REMARCA program and a control group of 314 healthy blood donors without autoimmune diseases and their presence in the history, who were matched with the study group for gender and sex. The polymorphic variants of the genes PTPN22 (+1858C>T, rs2476601), TNFAIP3 (rs6920220, rs10499194), CTLA4 (+49A>G, rs231775), TNFА (-308A>G, rs1800629), IL6 (-174G>C, rs1800795), IL6R (+358A>C, rs8192284), IL10 (-592A>C, rs1800872, -892 C>T, rs1800871, -1082 A>G, rs1800896), and MCP1/CCL2 (+2518A>G, rs1024611) were genotyped by a real-time polymerase chain reaction assay.

Results and discussion. The genotype and allele frequencies of polymorphic variants of the genes CTLA4 (+49A>G), IL-6R (+358A>C), and IL10 (592A>C) in the RA group significantly differed from those in the control group. When comparing with the control group, the minor alleles of the CTLA4 and IL10 genes were markers for the risk of aCCP-positive RA with a high inflammatory activity (OR=1.4 [1.1; 1.9], p=0.02 and OR=1.9 [1.4; 2.5]; p=0.0001, respectively). At the same time, the minor C allele of the IL6R gene served as a marker of protection (OR=0.7 [0.5; 0.9]; p=0.03). Logistic regression analysis revealed that there was a statistically significant correlation of the high inflammatory activity indices SDAI, CDAI, and DAS28 with the minor homozygous GG genotype of the CTLA4 gene (OR=2.5 [1.1; 6.0]; p=0.03, OR=2.6 [1.1–6.4], p=0.03 and OR=3.4 [1.3–8.8]; p=0.01, respectively). In addition, the inflammatory activity indices SDAI and CDAI rather than DAS28-ESR were associated with at least one minor A allele (the AA/AC genotypes) of the IL10 gene (OR=2.4 [1.2; 5.1], p=0.02 and OR=2.2 [1.1; 4.7]; p=0.03, respectively). The levels of ESR and CRP were not associated with the examined polymorphisms.

Conclusion. The findings may suggest that there is a relationship of the polymorphisms of the genes CTLA4 (+49A>G, rs231775), IL6R (+358A>C, rs8192284), and IL10 (-592A>C, rs1800872) to high inflammatory activity in the group of aCCP-positive patients from the REMARCA study. 

1. Насонов ЕЛ, Каратеев ДЕ, Балабанова РМ. Ревматоидный артрит. В кн.: Насонов ЕЛ, Насонова ВА, редакторы. Ревматология. Национальное руководство. Москва: ГЭОТАР-Медиа; 2008. С. 290-331 [Nasonov EL, Karateev DE, Balabanova RM. Rheumatoid arthritis. In: Nasonov EL, Nasonova VA, editors. Revmatologiya. Natsional'noe rukovodstvo [Rheumatology. National guidelance]. Moscow: GEOTARMedia; 2008. P. 290-331].

2. Van der Helm-van Mil AH, Huizinga TW. Advances in the genetics of rheumatoid arthritis point to subclassification into distinct disease subsets. Arthritis Res Ther. 2008;10(2):205. doi: 10.1186/ar2384

3. Padyukov L, Seielstad M, Ong RT, et al. A genome-wide association study suggests contrasting associations in ACPA-positive versus ACPA-negative rheumatoid arthritis. Ann Rheum Dis. 2011;70(2):259-65. doi: 10.1136/ard.2009. 126821

4. Гусева ИА, Демидова НВ, Сорока НЕ и др. Молекулярногенетическая характеристика раннего ревматоидного артрита. Молекулярная медицина. 2016;14(1):15-21 [Guseva IA, Demidova NV, Soroka NE, et al. Molecular-genetic characteristics of early rheumatoid arthritis. Molekulyarnaya Meditsina. 2016;14(1):15-21 (In Russ.)].

5. Гусева ИА, Демидова НВ, Сорока НЕ и др. Исследование полиморфизмов генов-кандидатов иммунного ответа как маркеров риска развития ревматоидного артрита и продукции аутоантител. Научно-практическая ревматология. 2016;54(1):21-30 [Guseva IA, Demidova NV, Soroka NE, et al. Investigation of candidate gene polymorphisms in an immune response as markers for the risk of developing rheumatoid arthritis and producing autoantibodies. NauchnoPrakticheskaya Revmatologiya = Rheumatology Science and Practice. 2016;54(1):21-30 (In Russ.)]. doi: 10.14412/1995-4484-2016-21-30

6. Кофиади ИА, Ребриков ДВ. Методы детекции однонуклеотидных полиморфизмов: аллель-специфическая ПЦР и гибридизация с олигонуклеотидной пробой. Генетика. 2006;42:22-32 [Kofiadi IA, Rebrikov DV. Detection methods for single nucleotide polymorphisms: allele-specific PCR and hybridization with oligonucleotide probe. Genetika. 2006;42:22-32 (In Russ.)].

7. Seidl C, Donner H, Fischer B, et al. CTLA4 codon 17 dimorphism in patients with rheumatoid arthritis. Tissue Antigens. 1998;51(1):62-6. doi: 10.1111/j.1399-0039.1998.tb02947.x

8. Ligers A, Teleshova N, Masterman T, et al. CTLA-4 gene expression is influenced by promoter and exon 1 polymorphisms. Genes Immun. 2001;2:145-52. doi: 10.1038/sj.gene.6363752

9. Mäurer M, Loserth S, Kolb-Maurer A, et al. A polymorphism in the human cytotoxic T-lymphocyte antigen 4 (CTLA4) gene (exon 1 +49) alters T cell activation. Immunogenetics. 2002;54:1-8. doi: 10.1007/s00251-002-0429-9

10. Han S, Li Y, Mao Y, Xie Y. Meta-analysis of the association of CTLA-4 exon-1 +49A/G polymorphism with rheumatoid arthritis. Hum Genet. 2005;118(1):123-32. doi: 10.1007/s00439-0050033-9

11. Lee YH, Bae SC, Choi SJ, et al. Association between the CTLA-4 +49 A/G polymorphism and susceptibility to rheumatoid arthritis: a meta-analysis. Mol Biol Rep. 2012;39(5):5599-605. doi: 10.1007/s11033-011-1364-3

12. Wang K, Zhu Q, Lu Y, et al. CTLA-4 +49 G/A Polymorphism Confers Autoimmune Disease Risk: An Updated Meta-Analysis. Genet Test Mol Biomarkers. 2017;21(4):222-7. doi: 10.1089/gtmb.2016.0335

13. Гусева ИА, Насонов ЕЛ. Генетика ревматоидного артрита. В кн.: Насонов ЕЛ, редактор. Генно-инженерные биологические препараты в лечении ревматоидного артрита. Москва: ИМА-ПРЕСС; 2013. С. 47-64 [Guseva IA, Nasonov EL. Genetics of rheumatoid arthritis. In: Nasonov EL, editor. Genno-inzhenernye biologicheskie preparaty v lechenii revmatoidnogo artrita [Genetically engineered biological agents in the treatment of rheumatoid arthritis]. Moscow: IMA-PRESS; 2013. P. 47-64].

14. Geng R, Song F, Yang X, et al. Association between cytotoxic T lymphocyte antigen-4 +49A/G, -1722T/C, and -1661A/G polymorphisms and cancer risk: a meta-analysis. Tumour Biol. 2014;35(4):3627-39. doi: 10.1007/s13277-013-1480-x

15. Ciccacci C, Conigliaro P, Perricone C, et al. Polymorphisms in STAT-4, IL-10, PSORS1C1, PTPN2 and MIR146A genes are associated differently with prognostic factors in Italian patients affected by rheumatoid arthritis. Clin Exp Immunol. 2016;186(2):157-63. doi: 10.1111/cei.12831

16. Paradowska-Gorycka A, Treflr J, Maciejewska-Stelmach J, Lacki JK. Interleukin-10 gene promoter polymorphism in Polish rheumatoid arthritis patients. Int J Immunogenet. 2010;37:225-31. doi: 10.1111/j.1744-313X.2010.00913.x

17. Ying B, Shi Y, Pan X, et al. Association of polymorphisms in the human IL-10 and IL-18 genes with rheumatoid arthritis. Mol Biol Rep. 2011;38:379-85. doi: 10.1007/s11033-010-0119-x

18. Гусева ИА, Панасюк ЕЮ, Сорока НЕ и др. Ассоциативная взаимосвязь генетических маркеров с эффективностью лечения ревматоидного артрита тоцилизумабом. Научно-практическая ревматология. 2013;51(4):377-82 [Guseva IA, Panasyuk EYu, Soroka NE, et al. Association of genetic markers with the efficiency of tocilizumab treatment for rheumatoid arthritis. NauchnoPrakticheskaya Revmatologiya = Rheumatology Science and Practice. 2013;51(4):377-82 (In Russ.)]. doi: 10.14412/1995-4484-2013-1247

19. Guseva IA, Soroka NE, Devyataykina AY, et al. TNFAIP3 rs675520 variant may predict response to rituximab treatment in rheumatoid arthritis. Ann Rheum Dis. 2012;71 Suppl 3:669.

20. Galicia JC, Tai H, Komatsu Y, et al. Polymorphisms in the IL-6 receptor (IL-6R) gene: strong evidence that serum levels of soluble IL-6R are genetically influenced. Genes Immun. 2004;5:513-6. doi: 10.1038/sj.gene.6364120

21. Marinou I, Walters K, Winfield J, et al. A gain of function polymorphism in the interleukin 6 receptor influences RA susceptibility. Ann Rheum Dis. 2010;69:1191-4. doi: 10.1136/ard.2008.100644

22. Rodriguez-Rodriguez L, Ramon Lamas J, Varade J, et al. Plasma soluble IL-6 receptor concentration in rheumatoid arthritis: associations with the rs8192284 IL6R polymorphism and with disease activity. Rheumatol Int. 2011;31:409-13. doi: 10.1007/s00296-0101593-0