Skip to main content

Advertisement

SpringerLink
Log in

MicroRNA-132, miR-146a, and miR-155 as potential biomarkers of methotrexate response in patients with rheumatoid arthritis

  • Original Article
  • Published:
Clinical Rheumatology Aims and scope Submit manuscript

Abstract

Introduction

Rheumatoid arthritis (RA) patients have high expression levels of hsa-miR-132-3p, hsa-miR-146a-5p, and hsa-miR-155-5p in peripheral blood. We studied if baseline blood levels of these microRNAs (miRNAs) could predict response to methotrexate (MTX).

Methods

RA patients (the American College of Rheumatology (ACR)/European League Against Rheumatism (EULAR) criteria) with active disease (disease-modifying anti-rheumatic drug (DMARD)–naïve and Disease Activity Score 28 (DAS28) > 3.2) were enrolled. They were treated with MTX by gradually increasing dose up to 25 mg/week. After 4 months, the DAS28 score was calculated and EULAR response was assessed. The hsa-miR-132-3p, hsa-miR-146a-5p, and hsa-miR-155-5p levels were measured by real-time qPCR in whole-blood RNA at baseline and 4 months after therapy, using hsa-let-7a-5p as housekeeping gene. Results are expressed as median (interquartile range).

Results

The 94 enrolled patients (81 females) had a median age of 40 (17) years, disease duration of (24) months, and DAS28 4.61 (1.11). After 4 months of therapy, 73 were classified as responders and 21 as non-responders. Baseline levels of all three miRNAs were lower in responders than non-responders, hsa-miR-132-3p (− 8.03 (0.70) versus − 7.47 (0.85), P < 0.05), hsa-miR-146a-5p (− 5.11 (0.88) versus − 4.62 (0.90), P < 0.05), and hsa-miR-155-5p (− 7.59 (1.07) versus − 7 (0.72), P = 0.002). Receiver operating characteristic curve analysis showed that all three miRNAs were also good predictors of MTX response, showing the following values: hsa-miR-132-3p (area under curve (AUC) 0.756, P < 0.05), hsa-miR-146a-5p (AUC 0.760, P < 0.05), and hsa-miR-155-5p (AUC 0.728, P = 0.002).

Conclusion

hsa-miR-132-3p, hsa-miR-146a-5p, and hsa-miR-155-5p are potential biomarkers of responsiveness to MTX therapy.

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

Similar content being viewed by others

References

  1. Halilova KI, Brown EE, Morgan SL, Bridges SL Jr, Hwang MH, Arnett DK et al (2012) Markers of treatment response to methotrexate in rheumatoid arthritis: where do we stand? Int J Rheumatol 2012:978396

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  2. Donahue KE, Gartlehner G, Jonas DE, Lux LJ, Thieda P, Jonas BL, Hansen RA, Morgan LC, Lohr KN (2008) Systematic review: comparative effectiveness and harms of disease-modifying medications for rheumatoid arthritis. Ann Intern Med 148:124–134

    Article  PubMed  Google Scholar 

  3. McInnes IB, Schett G (2011) The pathogenesis of rheumatoid arthritis. N Engl J Med 365:2205–2219

    Article  PubMed  CAS  Google Scholar 

  4. Fabre S, Dupuy A, Dossat N, Guisset C, Cohen J, Cristol J et al (2008) Protein biochip array technology for cytokine profiling predicts etanercept responsiveness in rheumatoid arthritis. Clin Exp Immunol 153:188–195

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  5. Van Venrooij WJ, Van Beers JJ, Pruijn GJ (2008) Anti-CCP antibody, a marker for the early detection of rheumatoid arthritis. Ann N Y Acad Sci 1143:268–285

    Article  PubMed  CAS  Google Scholar 

  6. Ling S, Bluett J, Barton A (2018) Prediction of response to methotrexate in rheumatoid arthritis. Expert Rev Clin Immunol 14:419–429

    Article  PubMed  CAS  Google Scholar 

  7. Davidson-Moncada J, Papavasiliou FN, Tam W (2010) MicroRNAs of the immune system. Ann N Y Acad Sci 1183:183–194

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  8. Sode J, Krintel SB, Carlsen AL, Hetland ML, Johansen JS, Hørslev-Petersen K et al (2017) Plasma microRNA profiles in patients with early rheumatoid arthritis responding to adalimumab plus methotrexate vs methotrexate alone: a placebo-controlled clinical trial. J Rheumatol 45:53–61

    Article  PubMed  CAS  Google Scholar 

  9. Cuppen BV, Rossato M, Fritsch-Stork RD, Concepcion AN, Schenk Y, Bijlsma JW et al (2016) Can baseline serum microRNAs predict response to TNF-alpha inhibitors in rheumatoid arthritis? Arthritis Res Ther 18:189

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  10. Pauley KM, Satoh M, Chan AL, Bubb MR, Reeves WH, Chan EK (2008) Upregulated miR-146a expression in peripheral blood mononuclear cells from rheumatoid arthritis patients. Arthritis Res Ther 10:R101

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  11. Lagos D, Pollara G, Henderson S, Gratrix F, Fabani M, Milne RS et al (2010) miR-132 regulates antiviral innate immunity through suppression of the p300 transcriptional co-activator. Nat Cell Biol 12:513–519

    Article  PubMed  CAS  Google Scholar 

  12. Chen X, Ba Y, Ma L, Cai X, Yin Y, Wang K, Guo J, Zhang Y, Chen J, Guo X, Li Q, Li X, Wang W, Zhang Y, Wang J, Jiang X, Xiang Y, Xu C, Zheng P, Zhang J, Li R, Zhang H, Shang X, Gong T, Ning G, Wang J, Zen K, Zhang J, Zhang CY (2008) Characterization of microRNAs in serum: a novel class of biomarkers for diagnosis of cancer and other diseases. Cell Res 18:997–1006

    Article  PubMed  CAS  Google Scholar 

  13. Vigorito E, Perks KL, Abreu-Goodger C, Bunting S, Xiang Z, Kohlhaas S, Das PP, Miska EA, Rodriguez A, Bradley A, Smith KGC, Rada C, Enright AJ, Toellner KM, MacLennan ICM, Turner M (2007) microRNA-155 regulates the generation of immunoglobulin class-switched plasma cells. Immunity 27:847–859

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  14. Zhang J, Cheng Y, Cui W, Li M, Li B, Guo L (2014) MicroRNA-155 modulates Th1 and Th17 cell differentiation and is associated with multiple sclerosis and experimental autoimmune encephalomyelitis. J Neuroimmunol 266:56–63

    Article  PubMed  CAS  Google Scholar 

  15. Taganov KD, Boldin MP, Chang K-J, Baltimore D (2006) NF-κB-dependent induction of microRNA miR-146, an inhibitor targeted to signaling proteins of innate immune responses. Proc Natl Acad Sci 103:12481–12486

    Article  PubMed  CAS  Google Scholar 

  16. Duroux-Richard I, Jorgensen C, Apparailly F (2011) miRNAs and rheumatoid arthritis—promising novel biomarkers. Swiss Med Wkly 141:w13175

    PubMed  Google Scholar 

  17. Murata K, Yoshitomi H, Tanida S, Ishikawa M, Nishitani K, Ito H, Nakamura T (2010) Plasma and synovial fluid microRNAs as potential biomarkers of rheumatoid arthritis and osteoarthritis. Arthritis Res Ther 12:R86

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  18. Churov AV, Oleinik EK, Knip M (2015) MicroRNAs in rheumatoid arthritis: altered expression and diagnostic potential. Autoimmun Rev 14:1029–1037

    Article  PubMed  CAS  Google Scholar 

  19. Pauley KM, Cha S (2011) miRNA-146a in rheumatoid arthritis: a new therapeutic strategy. Immunotherapy 3:829–831

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  20. Abdul-Maksoud R, Sediq A, Kattaia A, Elsayed W, Ezzeldin N, Abdel Galil S, Ibrahem R (2017) Serum miR-210 and miR-155 expression levels as novel biomarkers for rheumatoid arthritis diagnosis. Br J Biomed Sci 74:209–213

    Article  PubMed  CAS  Google Scholar 

  21. Zhou Q, Haupt S, Kreuzer JT, Hammitzsch A, Proft F, Neumann C, Leipe J, Witt M, Schulze-Koops H, Skapenko A (2015) Decreased expression of miR-146a and miR-155 contributes to an abnormal Treg phenotype in patients with rheumatoid arthritis. Ann Rheum Dis 74:1265–1274

    Article  PubMed  CAS  Google Scholar 

  22. Su LC, Huang AF, Jia H, Liu Y, Xu WD (2017) Role of micro RNA-155 in rheumatoid arthritis. Int J Rheum Dis 20:1631–1637

    Article  PubMed  CAS  Google Scholar 

  23. Rajagopalan PR, Zhang Z, McCourt L, Dwyer M, Benkovic SJ, Hammes GG (2002) Interaction of dihydrofolate reductase with methotrexate: ensemble and single-molecule kinetics. Proc Natl Acad Sci 99:13481–13486

    Article  PubMed  CAS  Google Scholar 

  24. Aletaha D, Neogi T, Silman AJ, Funovits J, Felson DT, Bingham IIICO et al (2010) 2010 rheumatoid arthritis classification criteria: an American College of Rheumatology/European League Against Rheumatism collaborative initiative. Arthritis Rheum 62:2569–2581

    Article  PubMed  Google Scholar 

  25. Prevoo M, Van’T Hof MA, Kuper H, Van Leeuwen M, Van De Putte L, Van Riel P (1995) Modified disease activity scores that include twenty-eight-joint counts development and validation in a prospective longitudinal study of patients with rheumatoid arthritis. Arthritis Rheum 38:44–48

    Article  PubMed  CAS  Google Scholar 

  26. Duroux-Richard I, Pers Y-M, Fabre S, Ammari M, Baeten D, Cartron G et al (2014) Circulating miRNA-125b is a potential biomarker predicting response to rituximab in rheumatoid arthritis. Mediat Inflamm 2014(23):342524. https://doi.org/10.1155/2014/342524

  27. Castro-Villegas C, Pérez-Sánchez C, Escudero A, Filipescu I, Verdu M, Ruiz-Limón P, Aguirre MA, Jiménez-Gomez Y, Font P, Rodriguez-Ariza A, Peinado JR, Collantes-Estévez E, González-Conejero R, Martinez C, Barbarroja N, López-Pedrera C (2015) Circulating miRNAs as potential biomarkers of therapy effectiveness in rheumatoid arthritis patients treated with anti-TNFα. Arthritis Res Ther 17:49

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  28. Pritchard CC, Kroh E, Wood B, Arroyo JD, Dougherty KJ, Miyaji MM et al (2011) Blood cell origin of circulating microRNAs: a cautionary note for cancer biomarker studies. Cancer Prev Res 5:492–497

    Article  CAS  Google Scholar 

  29. Kirschner MB, Kao SC, Edelman JJ, Armstrong NJ, Vallely MP, van Zandwijk N, Reid G (2011) Haemolysis during sample preparation alters microRNA content of plasma. PLoS One 6:e24145

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  30. Häusler S, Keller A, Chandran P, Ziegler K, Zipp K, Heuer S et al (2010) Whole blood-derived miRNA profiles as potential new tools for ovarian cancer screening. Br J Cancer 103:693–700

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  31. Patnaik SK, Yendamuri S, Kannisto E, Kucharczuk JC, Singhal S, Vachani A (2012) MicroRNA expression profiles of whole blood in lung adenocarcinoma. PLoS One 7:e46045

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  32. Schrauder MG, Strick R, Schulz-Wendtland R, Strissel PL, Kahmann L, Loehberg CR, Lux MP, Jud SM, Hartmann A, Hein A, Bayer CM, Bani MR, Richter S, Adamietz BR, Wenkel E, Rauh C, Beckmann MW, Fasching PA (2012) Circulating micro-RNAs as potential blood-based markers for early stage breast cancer detection. PLoS One 7:e29770

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  33. Mookherjee N, El-Gabalawy HS (2013) High degree of correlation between whole blood and PBMC expression levels of miR-155 and miR-146a in healthy controls and rheumatoid arthritis patients. J Immunol Methods 400:106–110

    Article  PubMed  CAS  Google Scholar 

  34. Nahid MA, Yao B, Dominguez-Gutierrez PR, Kesavalu L, Satoh M, Chan EK (2012) Regulation of TLR2-mediated tolerance and cross-tolerance through IRAK4 modulation by miR-132 and miR-212. J Immunol 190:1250–1263

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  35. Fehri LF, Koch M, Belogolova E, Khalil H, Bolz C, Kalali B et al (2010) Helicobacter pylori induces miR-155 in T cells in a cAMP-Foxp3-dependent manner. PLoS One 5:e9500

    Article  CAS  Google Scholar 

  36. Peres RS, Liew FY, Talbot J, Carregaro V, Oliveira RD, Almeida SL, França RFO, Donate PB, Pinto LG, Ferreira FIS, Costa DL, Demarque DP, Gouvea DR, Lopes NP, Queiroz RHC, Silva JS, Figueiredo F, Alves-Filho JC, Cunha TM, Ferreira SH, Louzada-Junior P, Cunha FQ (2015) Low expression of CD39 on regulatory T cells as a biomarker for resistance to methotrexate therapy in rheumatoid arthritis. Proc Natl Acad Sci 112:2509–2514

    Article  PubMed  CAS  Google Scholar 

  37. Boldin MP, Taganov KD, Rao DS, Yang L, Zhao JL, Kalwani M, Garcia-Flores Y, Luong M, Devrekanli A, Xu J, Sun G, Tay J, Linsley PS, Baltimore D (2011) miR-146a is a significant brake on autoimmunity, myeloproliferation, and cancer in mice. J Exp Med 208:1189–1201

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  38. Liu J, Shi K, Chen M, Xu L, Hong J, Hu B, Yang X, Sun R (2015) Elevated miR-155 expression induces immunosuppression via CD39+ regulatory T-cells in sepsis patient. Int J Infect Dis 40:135–141

    Article  PubMed  CAS  Google Scholar 

  39. Kohlhaas S, Garden OA, Scudamore C, Turner M, Okkenhaug K, Vigorito E (2009) Cutting edge: the Foxp3 target miR-155 contributes to the development of regulatory T cells. J Immunol 182:2578–2582

    Article  PubMed  CAS  Google Scholar 

  40. Hong H, Yang H, Xia Y (2018) Circulating miR-10a as predictor of therapy response in rheumatoid arthritis patients treated with methotrexate. Curr Pharm Biotechnol 19:79–86

    Article  PubMed  CAS  Google Scholar 

  41. Saevarsdottir S, Wedren S, Seddighzadeh M et al (2011) Patients with early rheumatoid arthritis who smoke are less likely to respond to treatment with methotrexate and tumor necrosis factor inhibitors: observations from the Epidemiological Investigation of Rheumatoid Arthritis and the Swedish Rheumatology Register cohorts. Arthritis Rheum 63:26–36

    Article  PubMed  CAS  Google Scholar 

  42. Anderson JJ, Wells G, Verhoeven AC, Felson DT (2000) Factors predicting response to treatment in rheumatoid arthritis: the importance of disease duration. Arthritis Rheum 43:22–29

    Article  PubMed  CAS  Google Scholar 

  43. O’Dell JR, Nepom BS, Haire C, Gersuk VH, Gaur L, Moore GF, Drymalski W, Palmer W, Eckhoff PJ, Klassen LW, Wees S, Thiele G, Nepom GT (1998) HLA-DRB1 typing in rheumatoid arthritis: predicting response to specific treatments. Ann Rheum Dis 57:209–213

    Article  PubMed  PubMed Central  Google Scholar 

  44. Qiu Q, Huang J, Shu X, Fan H, Zhou Y, Xiao C (2017) Polymorphisms and pharmacogenomics for the clinical efficacy of methotrexate in patients with rheumatoid arthritis: a systematic review and meta-analysis. Sci Rep 7:44015

    Article  PubMed  PubMed Central  Google Scholar 

  45. Drozdzik M, Rudas T, Pawlik A, Gornik W, Kurzawski M, Herczynska M (2007) Reduced folate carrier-1 80G>A polymorphism affects methotrexate treatment outcome in rheumatoid arthritis. Pharmacogenomics J 7:404–407

    Article  PubMed  CAS  Google Scholar 

  46. Nair VS, Pritchard CC, Tewari M, Ioannidis JP (2014) Design and analysis for studying microRNAs in human disease: a primer on -omic technologies. Am J Epidemiol 180:140–152

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Funding statement

The project was funded by a research grant to AA and AS was supported by the Senior Research Fellowship of Indian Council of Medical research.

Author information

Authors and Affiliations

  1. Department of Clinical Immunology and Rheumatology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, 226014, India

    Ankita Singh, Pradeepta Sekhar Patro & Amita Aggarwal

Authors
  1. Ankita Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Pradeepta Sekhar Patro
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Amita Aggarwal
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Amita Aggarwal.

Ethics declarations

Disclosures

None.

Additional information

Publisher’s Note

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

Electronic supplementary material

Supplementary Figure 1

Expression level of hsa-miR-132-3p (a), hsa-miR-146a-5p (b) and hsa-miR-155-5p (c) in responders at baseline (n=73) and 4 months after MTX therapy (n-66). Expression level of hsa-miR-132-3p (d), hsa-miR-146a-5p (e) and hsa-miR-155-5p (f) in non-responders at baseline (n=21) and 4 months after MTX therapy (n=14). *P < 0.05, as determined by Mann-Whitney U-test. BL, Baseline; FU, Follow-up; ∆Ct, Delta Threshold Cycle; R, Responder; NR, Non-Responder. (PDF 119 kb)

Supplementary Table 1

(PDF 251 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Singh, A., Patro, P.S. & Aggarwal, A. MicroRNA-132, miR-146a, and miR-155 as potential biomarkers of methotrexate response in patients with rheumatoid arthritis. Clin Rheumatol 38, 877–884 (2019). https://doi.org/10.1007/s10067-018-4380-z

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10067-018-4380-z

Keywords

Search

Navigation