Evaluation of ERAP1 Gene Single Nucleotide Polymorphism in Impressing the Inflammatory Cytokine Profile of Ankylosing Spondylitis Patients
Ankylosing spondylitis (AS), an autoinflammatory disease, has been associated with impaired Endoplasmic reticulum aminopeptidase (ERAP) 1 activity, which is involved in priming antigenic peptides. The purpose of this study was to evaluate if the genetic variant of ERAP1 gene could impress the inflammation status of the AS patients. For genotyping, 140 AS cases and 140 healthy controls were enrolled. After isolation of peripheral blood mononuclear cells (PBMCs) and DNA extraction, all the subjects were genotyped for rs27044 polymorphism using SSP-PCR assay. Total RNA of PBMCs was isolated, cDNA was synthesized, and quantitative analyses of mRNA expression of cytokines were performed via Real-time PCR using the SYBR Green Gene Expression MasterMix. To measure the concentration of cytokines in serum of subjects, Enzyme-linked immunosorbent assay (ELISA) was used. It was observed that the G allele of rs27044 polymorphism was significantly prevalent in AS patients. Moreover, the GG genotype and the GG+GC dominant model had significantly different distribution between study groups. There was a significant overexpression of mRNAs of IL-17A, IL-6, IL-33, TNF-α, and IFN-γ, while IL-10 was significantly downregulated in AS patients. The ELISA results were in line with that of the gene expression analysis. No significant differences in mRNA expression and concentration of cytokine were identified among AS patients with three genotypes for rs27044 SNP. This study replicated the association of polymorphisms in ERAP1 gene with the risk of AS in a population from Iranian. However, it did not directly determine the inflammatory profile of the AS patients.
1. Brown M, Wordsworth B, Reveille J. Genetics of ankylosing spondylitis. Clinical and experimental rheumatology. 2002;20(6; SUPP/28):S-43.
2. Mahmoudi M, Aslani S, Nicknam MH, Karami J, Jamshidi AR. New insights toward the pathogenesis of ankylosing spondylitis; genetic variations and epigenetic modifications. Modern rheumatology. 2017;27(2):198-209.
3. Mohammadi H, Sharafkandi N, Hemmatzadeh M, Azizi G, Karimi M, Jadidi-Niaragh F, et al. The role of innate lymphoid cells in health and disease. Journal of cellular physiology. 2018;233(6):4512-29.
4. Mohammadi H, Hemmatzadeh M, Babaie F, Gowhari Shabgah A, Azizi G, Hosseini F, et al. MicroRNA implications in the etiopathogenesis of ankylosing spondylitis. Journal of cellular physiology. 2018;233(8):5564-73.
5. Brown MA. Genetics and the pathogenesis of ankylosing spondylitis. Current opinion in rheumatology. 2009;21(4):318-23.
6. Brown MA, Kennedy LG, Darke C, Gibson K, Pile KD, Shatford JL, et al. The effect of HLA-DR genes on susceptibility to and severity of ankylosing spondylitis. Arthritis and rheumatism. 1998;41(3):460-5.
7. Australo-Anglo-American Spondyloarthritis C, Reveille JD, Sims AM, Danoy P, Evans DM, Leo P, et al. Genome-wide association study of ankylosing spondylitis identifies non-MHC susceptibility loci. Nature genetics. 2010;42(2):123-7.
8. Wang W, Meng X, Liu Y, Ma X, Zhang Q, Li C, et al. Association Between Protein Tyrosine Phosphatase Non-Receptor Type 22 (PTPN22) Polymorphisms and Risk of Ankylosing Spondylitis: A Meta-analysis. Medical science monitor : international medical journal of experimental and clinical research. 2017;23:2619-24.
9. Karaderi T, Keidel S, Pointon J, Appleton L, Brown M, Evans D, et al. Ankylosing spondylitis is associated with the anthrax toxin receptor 2 gene (ANTXR2). Annals of the rheumatic diseases. 2014;73(11):2054-8.
10. Rahman P, Inman RD, Gladman DD, Reeve JP, Peddle L, Maksymowych WP. Association of interleukin-23 receptor variants with ankylosing spondylitis. Arthritis and rheumatism. 2008;58(4):1020-5.
11. Harvey D, Pointon JJ, Evans DM, Karaderi T, Farrar C, Appleton LH, et al. Investigating the genetic association between ERAP1 and ankylosing spondylitis. Human molecular genetics. 2009;18(21):4204-12.
12. Jadon D, Tillett W, Wallis D, Cavill C, Bowes J, Waldron N, et al. Exploring ankylosing spondylitis-associated ERAP1, IL23R and IL12B gene polymorphisms in subphenotypes of psoriatic arthritis. Rheumatology. 2012;52(2):261-6.
13. Babaie F, Hasankhani M, Mohammadi H, Safarzadeh E, Rezaiemanesh A, Salimi R, et al. The role of gut microbiota and IL-23/IL-17 pathway in ankylosing spondylitis immunopathogenesis: New insights and updates. Immunology letters. 2018;196:52-62.
14. Fowler NL, Frazer IH. Mutations in TAP genes are common in cervical carcinomas. Gynecologic oncology. 2004;92(3):914-21.
15. Gostout BS, Poland GA, Calhoun ES, Sohni YR, Giuntoli Ii RL, McGovern RM, et al. TAP1, TAP2, and HLA-DR2 alleles are predictors of cervical cancer risk☆. Gynecologic oncology. 2003;88(3):326-32.
16. Saric T, Chang S-C, Hattori A, York IA, Markant S, Rock KL, et al. An IFN-γ–induced aminopeptidase in the ER, ERAP1, trims precursors to MHC class I–presented peptides. Nature immunology. 2002;3(12):1169.
17. Cui X, Hawari F, Alsaaty S, Lawrence M, Combs CA, Geng W, et al. Identification of ARTS-1 as a novel TNFR1-binding protein that promotes TNFR1 ectodomain shedding. The Journal of clinical investigation. 2002;110(4):515-26.
18. Cui X, Rouhani FN, Hawari F, Levine SJ. Shedding of the type II IL-1 decoy receptor requires a multifunctional aminopeptidase, aminopeptidase regulator of TNF receptor type 1 shedding. The Journal of Immunology. 2003;171(12):6814-9.
19. Chen R, Yao L, Meng T, Xu W. The association between seven ERAP1 polymorphisms and ankylosing spondylitis susceptibility: a meta-analysis involving 8,530 cases and 12,449 controls. Rheumatology international. 2012;32(4):909-14.
20. Cinar M, Akar H, Yilmaz S, Simsek I, Karkucak M, Sagkan RI, et al. A polymorphism in ERAP1 is associated with susceptibility to ankylosing spondylitis in a Turkish population. Rheumatology international. 2013;33(11):2851-8.
21. Tang Y, Yang P, Wang F, Xu H, Zong SY. Association of polymorphisms in ERAP1 and risk of ankylosing spondylitis in a Chinese population. Gene. 2018;646:8-11.
22. Burton PR, Clayton DG, Cardon LR, Craddock N, Deloukas P, Duncanson A, et al. Association scan of 14,500 nonsynonymous SNPs in four diseases identifies autoimmunity variants. Nature genetics. 2007;39(11):1329.
23. Davidson SI, Wu X, Liu Y, Wei M, Danoy PA, Thomas G, et al. Association of ERAP1, but not IL23R, with ankylosing spondylitis in a Han Chinese population. Arthritis & Rheumatology. 2009;60(11):3263-8.
24. Yong Y, Lin H. SHEsis, a powerful software platform for analyses of linkage disequilibrium, haplotype construction, and genetic association at polymorphism loci. Cell research. 2005;15(2):97-8.
25. Brown MA, Kenna T, Wordsworth BP. Genetics of ankylosing spondylitis—insights into pathogenesis. Nature Reviews Rheumatology. 2016;12(2):81.
26. Brown MA, Kennedy LG, MacGregor AJ, Darke C, Duncan E, Shatford JL, et al. Susceptibility to ankylosing spondylitis in twins: the role of genes, HLA, and the environment. Arthritis and rheumatism. 1997;40(10):1823-8.
27. Hamersma J, Cardon LR, Bradbury L, Brophy S, van der Horst-Bruinsma I, Calin A, et al. Is disease severity in ankylosing spondylitis genetically determined? Arthritis and rheumatism. 2001;44(6):1396-400.
28. Brionez TF, Reveille JD. The contribution of genes outside the major histocompatibility complex to susceptibility to ankylosing spondylitis. Current opinion in rheumatology. 2008;20(4):384-91.
29. Cortes A, Hadler J, Pointon JP, Robinson PC, Karaderi T, Leo P, et al. Identification of multiple risk variants for ankylosing spondylitis through high-density genotyping of immune-related loci. Nature genetics. 2013;45(7):730.
30. Evans DM, Spencer CC, Pointon JJ, Su Z, Harvey D, Kochan G, et al. Interaction between ERAP1 and HLA-B27 in ankylosing spondylitis implicates peptide handling in the mechanism for HLA-B27 in disease susceptibility. Nature genetics. 2011;43(8):761.
31. Reeves E, Elliott T, James E, Edwards CJ. ERAP1 in the pathogenesis of ankylosing spondylitis. Immunologic research. 2014;60(2-3):257-69.
32. Evnouchidou I, Momburg F, Papakyriakou A, Chroni A, Leondiadis L, Chang SC, et al. The internal sequence of the peptide-substrate determines its N-terminus trimming by ERAP1. PloS one. 2008;3(11):e3658.
33. Mahmoudi M, Jamshidi AR, Amirzargar AA, Farhadi E, Nourijelyani K, Fallahi S, et al. Association between endoplasmic reticulum aminopeptidase-1 (ERAP-1) and susceptibility to ankylosing spondylitis in Iran. Iranian Journal of Allergy, Asthma and Immunology. 2012;11(4):294.
34. Lee YH, Choi SJ, Ji JD, Song GG. Associations between ERAP1 polymorphisms and ankylosing spondylitis susceptibility: a meta-analysis. Inflammation Research. 2011;60(11):999.
35. Ombrello MJ, Kastner DL, Remmers EF. Endoplasmic reticulum-associated amino-peptidase 1 and rheumatic disease: genetics. Current opinion in rheumatology. 2015;27(4):349-56.
36. Maksymowych WP, Inman RD, Gladman DD, Reeve JP, Pope A, Rahman P. Association of a specific ERAP1/ARTS1 haplotype with disease susceptibility in ankylosing spondylitis. Arthritis and rheumatism. 2009;60(5):1317-23.
37. Kadi A, Izac B, Said-Nahal R, Leboime A, Van Praet L, de Vlam K, et al. Investigating the genetic association between ERAP1 and spondyloarthritis. Annals of the rheumatic diseases. 2012:annrheumdis-2012-201783.
38. Bettencourt BF, Rocha FL, Alves H, Amorim R, Caetano-Lopes J, Vieira-Sousa E, et al. Protective effect of an ERAP1 haplotype in ankylosing spondylitis: investigating non-MHC genes in HLA-B27-positive individuals. Rheumatology. 2013;52(12):2168-76.
39. Haroon N, Maksymowych WP, Rahman P, Tsui FW, O'Shea FD, Inman RD. Radiographic severity of ankylosing spondylitis is associated with polymorphism of the large multifunctional peptidase 2 gene in the Spondyloarthritis Research Consortium of Canada cohort. Arthritis & Rheumatology. 2012;64(4):1119-26.