Skip to main content

Advertisement

SpringerLink
Log in

Genetic and expression deregulation of immunoregulatory genes in rheumatoid arthritis

  • Original Article
  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

Rheumatoid arthritis (RA) is one of the most common autoimmune diseases globally, and is an important public health concern, associating with early death and systemic complications. Although key development in RA treatment has already been made still RA affected individuals face comorbidity and disability. Therefore, there is a need to discover new risk factors in helping early diagnosis and treatment of RA. The present study is designed to assess the variations of Vitamin D receptor (VDR) and interleukin -6 (IL-6) in RA patients. Polymorphisms of said genes were calculated in 300 RA patients and 300 controls, using Tetra-ARMS polymerase chain reaction. Secondly, expression levels of selected genes were checked using the quantitative PCR (qPCR) and obtained results were evaluated using a different statistical test. Logistic regression analysis showed that frequency of mutant allele of VDR gene polymorphisms (rs11168268, OR = 4.84; 95% CI = 2.94–7.97; p = 0.0001; rs2248098, OR = 1.65; 95% CI = 1.07–2.54; p = 0.02) and IL-6 gene polymorphisms (rs184229712, OR = 2.47; 95% CI = 1.56–3.92, p = 0.0001; rs36215814, OR = 2.14; 95% CI = 1.30–3.53; p = 0.002) was observed significantly higher in RA patients vs controls. Expression analysis showed the significant upregulation of IL-6 (p < 0.0001) and downregulation of VDR gene (p < 0.0001) in RA cases vs controls. ROC curve analysis showed that downregulation of IL-6 (AUC = 0.86, p < 0.001) and upregulation of VDR (AUC = 0.77, p < 0.001) was act as the good diagnostic marker for detection/diagnosis of arthritis. In conclusions, data from the present study showed the significant involvement of VDR and IL-6 gene variations in RA pathogenesis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Data availability

All data generated or analyzed during this study are included in this published article.

Abbreviations

RA:

Rheumatoid Arthritis

VDR:

Vitamin D Receptor

IL-6:

Interleukin -6

Tetra-ARMS-PCR:

Tetra- Amplification-Refractory Mutation System Polymerase Chain Reaction

AUC:

Area Under Curve

VD:

Vitamin D

TAE:

Tris Acetic Acid EDTA

IDT:

Integrated DNA Technologies

CI:

Confidence Interval

ANOVA:

Analysis of Variance

ESR:

Erythrocyte Sedimentation Rate

Anti-CCP:

Cyclic Citrullinated Peptide Antibody

ESR:

Erythrocyte Sedimentation Rate

CRP:

C-Reactive Protein

References

  1. Hitchon C, El-Gabalawy H (2011) The synovium in rheumatoid arthritis. Open Rheumatol J 5:107–114

    Article  PubMed  PubMed Central  Google Scholar 

  2. Tobón GJ, Youinou P, Saraux A (2010) The environment, geo-epidemiology, and autoimmune disease: rheumatoid arthritis. Autoimmun Rev 9(5):A288–A292

    Article  PubMed  CAS  Google Scholar 

  3. Alamanos Y, Drosos AA (2005) Epidemiology of adult rheumatoid arthritis. Autoimmun Rev 4(3):130–136

    Article  PubMed  Google Scholar 

  4. Mukhtar M, Sheikh N, Suqaina SK, Batool A, Fatima N, Mehmood R, Nazir S (2019) Vitamin D receptor gene polymorphism: an important predictor of arthritis development. BioMed Res Int. https://doi.org/10.1155/2019/8326246

    Article  PubMed  PubMed Central  Google Scholar 

  5. Kerr GS, Sabahi I, Richards JS, Caplan L, Cannon GW, Reimold A, Thiele GM, Johnson D, Mikuls TR (2011) Prevalence of vitamin D insufficiency/deficiency in rheumatoid arthritis and associations with disease severity and activity. J Rheumatol 38(1):53–59

    Article  PubMed  Google Scholar 

  6. Gopinath K, Danda D (2011) Supplementation of 1, 25 dihydroxy vitamin D3 in patients with treatment naive early rheumatoid arthritis: a randomised controlled trial. Int J Rheum Dis 14(4):332–339

    Article  PubMed  Google Scholar 

  7. Furuya T, Hosoi T, Tanaka E, Nakajima A, Taniguchi A, Momohara S, Yamanaka H (2013) Prevalence of and factors associated with vitamin D deficiency in 4,793 Japanese patients with rheumatoid arthritis. Clin Rheumatol 32(7):1081–1087

    Article  PubMed  Google Scholar 

  8. Haga H-J, Schmedes A, Naderi Y, Moreno AM, Peen E (2013) Severe deficiency of 25-hydroxyvitamin D 3 (25-OH-D 3) is associated with high disease activity of rheumatoid arthritis. Clin Rheumatol 32(5):629–633

    Article  PubMed  Google Scholar 

  9. Cutolo M (2009) Vitamin D and autoimmune rheumatic diseases. Oxford University Press, Oxford

    Google Scholar 

  10. Dupuis ML, Pagano MT, Pierdominici M, Ortona E (2021) The role of vitamin D in autoimmune diseases: could sex make the difference? Biol Sex Differ 12(1):1–12

    Article  CAS  Google Scholar 

  11. Murdaca G, Tonacci A, Negrini S, Greco M, Borro M, Puppo F, Gangemi S (2019) Emerging role of vitamin D in autoimmune diseases: an update on evidence and therapeutic implications. Autoimmun Rev 18(9):102350

    Article  CAS  PubMed  Google Scholar 

  12. Zhou T-B, Jiang Z-P, Lin Z-J, Su N (2015) Association of vitamin D receptor gene polymorphism with the risk of systemic lupus erythematosus. J Recept Signal Transduction 35(1):8–14

    Article  CAS  Google Scholar 

  13. Tizaoui K, Hamzaoui K (2015) Association between VDR polymorphisms and rheumatoid arthritis disease: systematic review and updated meta-analysis of case–control studies. Immunobiology 220(6):807–816

    Article  CAS  PubMed  Google Scholar 

  14. Tizaoui K, Kaabachi W, Salah MO, Amor AB, Hamzaoui A, Hamzaoui K (2014) Vitamin D receptor TaqI and ApaI polymorphisms: a comparative study in patients with Behçet’s disease and Rheumatoid arthritis in Tunisian population. Cell Immunol 290(1):66–71

    Article  CAS  PubMed  Google Scholar 

  15. Ates O, Dolek B, Dalyan L, Topal-Sarikaya A (2011) Vitamin D receptor gene polymorphisms in rheumatoid arthritis/romatoid artritte d vitamini reseptoru geni polimorfizmleri. Turk J Rheumatol 26(2):145–150

    Article  Google Scholar 

  16. Shukla S, Tripathi AK, Tripathi JK, Indurkar M, Chauhan UK (2014) Role of PTPN22 and VDR gene polymorphisms in susceptibility to rheumatoid arthritis: a study from central India. Adv Genom Genet 4:79–85

    Article  CAS  Google Scholar 

  17. Saad MN, Mabrouk MS, Eldeib AM, Shaker OG (2015) Genetic case-control study for eight polymorphisms associated with rheumatoid arthritis. PloS one 10(7):e0131960

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  18. Hitchon CA, Sun Y, Robinson DB, Peschken CA, Bernstein CN, Siminovitch KA, El-Gabalawy HS (2012) Vitamin D receptor polymorphism rs2228570 (Fok1) is associated with rheumatoid arthritis in North American natives. J Rheumatol 39(9):1792–1797

    Article  CAS  PubMed  Google Scholar 

  19. Karray EF, Dhifallah IB, Abdelghani KB, Ghorbel IB, Khanfir M, Houman H, Hamzaoui K, Zakraoui L (2012) Associations of vitamin D receptor gene polymorphisms FokI and BsmI with susceptibility to rheumatoid arthritis and Behçet’s disease in Tunisians. Joint Bone Spine 79(2):144–148

    Article  CAS  PubMed  Google Scholar 

  20. Sun G, Ba C-L, Gao R, Liu W, Ji Q (2019) Association of IL-6, IL-8, MMP-13 gene polymorphisms with knee osteoarthritis susceptibility in the Chinese Han population. Biosci Rep. https://doi.org/10.1042/BSR20181346

  21. Sun G, Wu G, Meng Y, Du B, Li Y (2014) IL-6 gene promoter polymorphisms and risk of coronary artery disease in a Chinese population. Genet Mol Res 13(3):7718–7724

    Article  CAS  PubMed  Google Scholar 

  22. García-Martín E, Agúndez JA, Martínez C, Benito-León J, Millán-Pascual J, Calleja P, Díaz-Sánchez M, Pisa D, Turpín-Fenoll L, Alonso-Navarro H (2013) Vitamin D3 receptor (VDR) gene rs2228570 (Fok1) and rs731236 (Taq1) variants are not associated with the risk for multiple sclerosis: results of a new study and a meta-analysis. PloS one 8(6):e65487

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  23. Huang J, Xie Z-F (2012) Polymorphisms in the vitamin D receptor gene and multiple sclerosis risk: a meta-analysis of case–control studies. J Neurol Sci 313(1–2):79–85

    Article  CAS  PubMed  Google Scholar 

  24. Tizaoui K, Kaabachi W, Hamzaoui A, Hamzaoui K (2014) Contribution of VDR polymorphisms to type 1 diabetes susceptibility: systematic review of case–control studies and meta-analysis. J Steroid Biochem Mol Biol 143:240–249

    Article  CAS  PubMed  Google Scholar 

  25. Hussien YM, Shehata A, Karam RA, Alzahrani SS, Magdy H, El-Shafey AM (2013) Polymorphism in vitamin D receptor and osteoprotegerin genes in Egyptian rheumatoid arthritis patients with and without osteoporosis. Mol Biol Rep 40(5):3675–3680

    Article  CAS  PubMed  Google Scholar 

  26. Mahjabeen I, Baig RM, Masood N, Sabir M, Inayat U, Malik FA, Kayani MA (2013) Genetic variations in XRCC1 gene in sporadic head and neck cancer (HNC) patients. Pathol Oncol Res 19(2):183–188

    Article  CAS  PubMed  Google Scholar 

  27. Rio DC, Ares M, Hannon GJ, Nilsen TW (2010) Purification of RNA using TRIzol (TRI reagent). Cold Spring Harbor Protocols 2010(6):pdb.prot5439

    Article  PubMed  Google Scholar 

  28. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods 25(4):402–408

    Article  CAS  PubMed  Google Scholar 

  29. Barrett JC (2009) Haploview: visualization and analysis of SNP genotype data. Cold Spring Harbor Protocols 2009(10):pdb.ip71

    Article  PubMed  Google Scholar 

  30. Ahmed MW, Mahjabeen I, Gul S, Khursheed A, Mehmood A, Kayani MA (2019) Relationship of single nucleotide polymorphisms and haplotype interaction of mitochondrial unfolded protein response pathway genes with head and neck cancer. Future Oncol 15(33):3819–3829

    Article  CAS  PubMed  Google Scholar 

  31. Marinou I, Healy J, Mewar D, Moore D, Dickson M, Binks M, Montgomery D, Walters K, Wilson A (2007) Association of interleukin-6 and interleukin-10 genotypes with radiographic damage in rheumatoid arthritis is dependent on autoantibody status. Arthritis Rheum 56(8):2549–2556

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Jančić I, Arsenović-Ranin N, Šefik-Bukilica M, Živojinović S, Damjanov N, Spasovski V, Srzentić S, Stanković B, Pavlović S (2013) 174G/C interleukin-6 gene promoter polymorphism predicts therapeutic response to etanercept in rheumatoid arthritis. Rheumatol Int 33(6):1481–1486

    Article  PubMed  CAS  Google Scholar 

  33. Cavalcanti CAJ, de Azevêdo SJ, de Barros PW, Veit TD, Monticielo OA, Xavier RM, Brenol JCT, Brenol CV, Fragoso TS, Barbosa AD (2016) Vitamin D receptor polymorphisms and expression profile in rheumatoid arthritis brazilian patients. Mol Biol Rep 43(1):41–51

    Article  CAS  PubMed  Google Scholar 

  34. Pike JW, Meyer MB, Watanuki M, Kim S, Zella LA, Fretz JA, Yamazaki M, Shevde NK (2007) Perspectives on mechanisms of gene regulation by 1, 25-dihydroxyvitamin D3 and its receptor. J Steroid Biochem Mol Biol 103(3–5):389–395

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Haussler MR, Haussler CA, Bartik L, Whitfield GK, Hsieh J-C, Slater S, Jurutka PW (2008) Vitamin D receptor: molecular signaling and actions of nutritional ligands in disease prevention. Nutr Rev 66(suppl_2):S98–S112

    Article  PubMed  Google Scholar 

  36. Pike JW, Meyer MB (2012) The vitamin D receptor: new paradigms for the regulation of gene expression by 1, 25-dihydroxyvitamin D3. Rheum Dis Clin 38(1):13–27

    Article  Google Scholar 

  37. Garcia-Lozano J, Gonzalez-Escribano M, Valenzuela A, Garcia A, Nunez-Roldan A (2001) Association of vitamin D receptor genotypes with early onset rheumatoid arthritis. Eur J Immunogenet 28(1):89–93

    CAS  PubMed  Google Scholar 

  38. Maalej A, Petit-Teixeira E, Michou L, Rebai A, Cornélis F, Ayadi H (2005) Association study of VDR gene with rheumatoid arthritis in the French population. Genes Immun 6(8):707–711

    Article  CAS  PubMed  Google Scholar 

  39. Ad’hiah AH, Mahmood AS, Al-kazaz A-KA, Mayouf KK (2018) Gene expression and six single nucleotide polymorphisms of interleukin-6 in rheumatoid arthritis: a case-control study in Iraqi patients. Alex J Med 54(4):639–645

    Google Scholar 

  40. Zavaleta-Muñiz S, Martín-Márquez B, Gonzalez-Lopez L, Gonzalez-Montoya N, Díaz-Toscano M, Ponce-Guarneros J, Ruiz-Padilla A, Mercado M, Maldonado-González M, Fafutis-Morris M (2013) The− 174G/C and− 572G/C interleukin 6 promoter gene polymorphisms in Mexican patients with rheumatoid arthritis: a case-control study. Clin Dev Immunol 2013:1–5

    Article  CAS  Google Scholar 

  41. Palomino-Morales R, Gonzalez-Juanatey C, Vazquez-Rodriguez T, Miranda-Filloy J, Llorca J, Martin J, Gonzalez-Gay M (2009) Interleukin-6 gene-174 promoter polymorphism is associated with endothelial dysfunction but not with disease susceptibility in patients with rheumatoid arthritis. Clin Exp Rheumatol 27(6):964–970

    CAS  PubMed  Google Scholar 

  42. Lopez-Lasanta M, Julià A, Maymó J, Fernández-Gutierrez B, Ureña-Garnica I, Blanco FJ, Cañete JD, Alperi-López M, Olivè A, Corominas H (2015) Variation at interleukin-6 receptor gene is associated to joint damage in rheumatoid arthritis. Arthritis Res Ther 17(1):1–8

    Article  CAS  Google Scholar 

  43. Nishimura K, Sugiyama D, Kogata Y, Tsuji G, Nakazawa T, Kawano S, Saigo K, Morinobu A, Koshiba M, Kuntz KM (2007) Meta-analysis: diagnostic accuracy of anti–cyclic citrullinated peptide antibody and rheumatoid factor for rheumatoid arthritis. Ann Intern Med 146(11):797–808

    Article  PubMed  Google Scholar 

  44. Avouac J, Gossec L, Dougados M (2006) Diagnostic and predictive value of anti-cyclic citrullinated protein antibodies in rheumatoid arthritis: a systematic literature review. Ann Rheum Dis 65(7):845–851

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Bos WH, Wolbink GJ, Boers M, Tijhuis GJ, de Vries N, van der Horst-Bruinsma IE, Tak PP, Van De Stadt R, van der Laken CJ, Dijkmans BA (2010) Arthritis development in patients with arthralgia is strongly associated with anti-citrullinated protein antibody status: a prospective cohort study. Ann Rheum Dis 69(3):490–494

    Article  CAS  PubMed  Google Scholar 

  46. Bashir K, Sarwar R, Saeed S, Mahjabeen I, Kayani MA (2018) Interaction among susceptibility genotypes of PARP1 SNPs in thyroid carcinoma. PloS One 13(9):e0199007

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  47. Khan A, Mahjabeen I, Malik MA, Hussain MZ, Khan S, Kayani MA (2019) Modulation of brain tumor risk by genetic SNPs in PARP1gene: hospital based case control study. PloS One 14(10):e0223882

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

The authors are thankful to patients and staff of the military hospital (MH), Rawalpindi, for their contribution to this research.

Author information

Authors and Affiliations

  1. Department of Rheumatology, National University of Medical Sciences, Rawalpindi, Pakistan

    Muhammad Zhaid Hussain, Naila Mumtaz, Syed Uzair Maqsood, Farooq Ikram, Qurrat-ul-Ain Kalim & Rabia Abbas

  2. Cancer Genetics and Epigenetics Lab, Department of Biosciences, COMSATS University, Islamabad, Pakistan

    Ishrat Mahjabeen

  3. HITEC-Institute of Medical Sciences, Rawalpindi, Pakistan

    Muhammad Shahid Khan

  4. Department of Rheumatology, Fauji Foundation Hospital, Rawalpindi, Pakistan

    Syed Nazir Ahmed

  5. Department of Medicine, Sheikh Zayed Medical College, and Hospital, Rahim Yar Khan, Pakistan

    Ahmed Ammar Cheema

Authors
  1. Muhammad Zhaid Hussain
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ishrat Mahjabeen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Muhammad Shahid Khan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Naila Mumtaz
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Syed Uzair Maqsood
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Farooq Ikram
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Syed Nazir Ahmed
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Qurrat-ul-Ain Kalim
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Rabia Abbas
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Ahmed Ammar Cheema
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

All authors contributed to the study’s conception and design. Material preparation and data collection were performed by [MSK], [NM], [AAC], and [SUM]. Experimentation and analysis were performed by [FI], [SNA], [Q-u-AK], and [RA]. The first draft of the manuscript was written by [MZH] and [IM] and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Muhammad Zhaid Hussain.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interests.

Consent to participate

All authors have seen the manuscript and approved to submit the manuscript.

Consent for publication

All authors consent to the publication of the manuscript. Furthermore, consent from cases and controls was obtained to publish their data before submitting their paper to a journal.

Ethical approval

Present study was conducted after the prior approval by the Ethical Review Committee of the Military hospital, Rawalpindi (RTMC # RHE-2019-124-87). The procedures used in this study adhere to the tenets of the Declaration of Helsinki.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Hussain, M.Z., Mahjabeen, I., Khan, M.S. et al. Genetic and expression deregulation of immunoregulatory genes in rheumatoid arthritis. Mol Biol Rep 48, 5171–5180 (2021). https://doi.org/10.1007/s11033-021-06518-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-021-06518-3

Keywords

Search

Navigation