Integrated Analysis of Circulating miR-22-3p, miR-126-3p, and miR-760 as Non-invasive Biomarkers for Rheumatoid Arthritis
-
Alireza Raghibi
,
Amirhossein Mohajeri Khorasani
,
Elia Damavandi
,
Abdolrahman Rostamian
,
Mozhgan Abjoushak Mahmoudabad
,
Hamid Choobineh
,
Mohsen Ghadami
,
Majid Kabuli
Abstract
Rheumatoid arthritis (RA) is an autoimmune disease marked by chronic inflammation and progressive joint damage. Early diagnosis is crucial, but current methods lack sensitivity. Circulating microRNAs (miRs), stable in body fluids, have emerged as promising biomarkers. This study aims to evaluate the plasma expression levels of miR-22-3p, miR-126-3p, and miR-760 in patients with RA compared to healthy controls, and to assess their potential as diagnostic biomarkers.
Plasma samples from 50 RA patients and 50 healthy individuals were analyzed using quantitative real-time PCR (qRT-PCR). Correlation of miR expression with disease activity was evaluated, and their diagnostic value was assessed using receiver operating characteristic (ROC) curve analysis. Bioinformatics analysis was conducted to explore the various aspects and functions of the identified miRs.
The plasma levels of miR-22-3p and miR-126-3p were significantly elevated in RA patients compared to healthy controls. Although miR-760 exhibited an upward trend, the increase did not reach statistical significance. miR-126-3p was positively associated with disease activity. ROC analysis showed area under the curve (AUC) of 0.69 for miR-22-3p, 0.72 for miR-126-3p, and 0.61 for miR-760, with the combined panel improving diagnostic accuracy to an AUC of 0.74. Furthermore, functional enrichment analysis suggested that these miRs are predominantly involved in key biological processes, including regulation of gene expression, cell migration, and epigenetic modifications.
A panel consisting of miR-22-3p, miR-126-3p, and miR-760 may serve as a potential diagnostic biomarker for RA. Further validation in larger and more diverse populations is warranted.
Keywords:;; ; ; ;
1. Bullock J, Rizvi SAA, Saleh AM, Ahmed SS, Do DP, Ansari RA, et al. Rheumatoid Arthritis: A Brief Overview of the Treatment. Med Princ Pract. 2018;27(6):501-7.
2. Venetsanopoulou AI, Alamanos Y, Voulgari PV, Drosos AA. Epidemiology and Risk Factors for Rheumatoid Arthritis Development. Mediterr J Rheumatol. 2023;34(4):404-13.
3. Scherer HU, Häupl T, Burmester GR. The etiology of rheumatoid arthritis. Journal of Autoimmunity. 2020;110:102400.
4. Jang S, Kwon EJ, Lee JJ. Rheumatoid Arthritis: Pathogenic Roles of Diverse Immune Cells. Int J Mol Sci. 2022;23(2).
5. Cheng Q, Chen X, Wu H, Du Y. Three hematologic/immune system-specific expressed genes are considered as the potential biomarkers for the diagnosis of early rheumatoid arthritis through bioinformatics analysis. J Transl Med. 2021;19(1):18.
6. O'Brien J, Hayder H, Zayed Y, Peng C. Overview of MicroRNA Biogenesis, Mechanisms of Actions, and Circulation. Front Endocrinol (Lausanne). 2018;9:402.
7. Pozniak T, Shcharbin D, Bryszewska M. Circulating microRNAs in Medicine. Int J Mol Sci. 2022;23(7).
8. Wu Y, Li Q, Zhang R, Dai X, Chen W, Xing D. Circulating microRNAs: Biomarkers of disease. Clin Chim Acta. 2021;516:46-54.
9. Peng X, Wang Q, Li W, Ge G, Peng J, Xu Y, et al. Comprehensive overview of microRNA function in rheumatoid arthritis. Bone Res. 2023;11(1):8.
10. Kent WJ, Sugnet CW, Furey TS, Roskin KM, Pringle TH, Zahler AM, et al. The human genome browser at UCSC. Genome research. 2002;12(6):996-1006.
11. Nassar LR, Barber GP, Benet-Pagès A, Casper J, Clawson H, Diekhans M, et al. The UCSC Genome Browser database: 2023 update. Nucleic Acids Research. 2023;51(D1):D1188-D95.
12. Kozomara A, Birgaoanu M, Griffiths-Jones S. miRBase: from microRNA sequences to function. Nucleic Acids Res. 2019;47(D1):D155-d62.
13. Zuker M. Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Research. 2003;31(13):3406-15.
14. Ru Y, Kechris KJ, Tabakoff B, Hoffman P, Radcliffe RA, Bowler R, et al. The multiMiR R package and database: integration of microRNA-target interactions along with their disease and drug associations. Nucleic Acids Res. 2014;42(17):e133.
15. R Core Team (2022). R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. URL https://www.R-project.org/.
16. RStudio Team (2022). RStudio: Integrated Development Environment for R. RStudio, PBC, Boston, MA URL http://www.rstudio.com/.
17. Li J-H, Liu S, Zhou H, Qu L-H, Yang J-H. starBase v2.0: decoding miRNA-ceRNA, miRNA-ncRNA and protein–RNA interaction networks from large-scale CLIP-Seq data. Nucleic Acids Research. 2013;42(D1):D92-D7.
18. Shannon P, Markiel A, Ozier O, Baliga NS, Wang JT, Ramage D, et al. Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Res. 2003;13(11):2498-504.
19. Chin C-H, Chen S-H, Wu H-H, Ho C-W, Ko M-T, Lin C-Y. cytoHubba: identifying hub objects and sub-networks from complex interactome. BMC Systems Biology. 2014;8(4):S11.
20. Xie Z, Bailey A, Kuleshov MV, Clarke DJ, Evangelista JE, Jenkins SL, et al. Gene set knowledge discovery with enrichr. Current protocols. 2021;1(3):e90.
21. H. Wickham. ggplot2: Elegant Graphics for Data Analysis. Springer-Verlag New York, 2016.
22. Zamani M, Cassiman JJ. Reevaluation of the importance of polymorphic HLA class II alleles and amino acids in the susceptibility of individuals of different populations to type I diabetes. American journal of medical genetics. 1998;76(2):183-94.
23. Davatchi F, Sandoughi M, Moghimi N, Jamshidi AR, Tehrani Banihashemi A, Zakeri Z, et al. Epidemiology of rheumatic diseases in Iran from analysis of four COPCORD studies. International journal of rheumatic diseases. 2016;19(11):1056-62.
24. Ding Q, Hu W, Wang R, Yang Q, Zhu M, Li M, et al. Signaling pathways in rheumatoid arthritis: implications for targeted therapy. Signal Transduct Target Ther. 2023;8(1):68.
25. Kim JW, Suh CH. Systemic Manifestations and Complications in Patients with Rheumatoid Arthritis. J Clin Med. 2020;9(6).
26. Steiner G, Verschueren P, Van Hoovels L, Studenic P, Bossuyt X. Classification of rheumatoid arthritis: is it time to revise the criteria? RMD Open. 2024;10(2).
27. Metcalf GAD. MicroRNAs: circulating biomarkers for the early detection of imperceptible cancers via biosensor and machine-learning advances. Oncogene. 2024;43(28):2135-42.
28. Gayosso-Gómez LV, Ortiz-Quintero B. Circulating MicroRNAs in Blood and Other Body Fluids as Biomarkers for Diagnosis, Prognosis, and Therapy Response in Lung Cancer. Diagnostics (Basel). 2021;11(3).
29. Evangelatos G, Fragoulis GE, Koulouri V, Lambrou GI. MicroRNAs in rheumatoid arthritis: From pathogenesis to clinical impact. Autoimmun Rev. 2019;18(11):102391.
30. Tang J, Lin J, Yu Z, Jiang R, Xia J, Yang B, et al. Identification of circulating miR-22-3p and let-7a-5p as novel diagnostic biomarkers for rheumatoid arthritis. Clin Exp Rheumatol. 2022;40(1):69-77.
31. Cieśla M, Kolarz B, Majdan M, Darmochwał-Kolarz D. Plasma micro-RNA-22 is associated with disease activity in well-established rheumatoid arthritis. Clin Exp Rheumatol. 2022;40(5):945-51.
32. Ormseth MJ, Solus JF, Sheng Q, Ye F, Song H, Wu Q, et al. The Endogenous Plasma Small RNAome of Rheumatoid Arthritis. ACR Open Rheumatol. 2020;2(2):97-105.
33. Renman E, Brink M, Ärlestig L, Rantapää-Dahlqvist S, Lejon K. Dysregulated microRNA expression in rheumatoid arthritis families-a comparison between rheumatoid arthritis patients, their first-degree relatives, and healthy controls. Clin Rheumatol. 2021;40(6):2387-94.
34. Cunningham CC, Wade S, Floudas A, Orr C, McGarry T, Wade S, et al. Serum miRNA Signature in Rheumatoid Arthritis and "At-Risk Individuals". Front Immunol. 2021;12:633201.
35. Ormseth MJ, Solus JF, Sheng Q, Chen SC, Ye F, Wu Q, et al. Plasma miRNAs improve the prediction of coronary atherosclerosis in patients with rheumatoid arthritis. Clin Rheumatol. 2021;40(6):2211-9.
36. Abdelaleem OO, Fouad NA, Shaker OG, Ahmed TI, Abdelghaffar NK, Eid HM, et al. Serum miR-224, miR-760, miR-483-5p, miR-378 and miR-375 as potential novel biomarkers in rheumatoid arthritis. Int J Clin Pract. 2021;75(11):e14651.
37. Lai NS, Yu HC, Tung CH, Huang KY, Huang HB, Lu MC. The role of aberrant expression of T cell miRNAs affected by TNF-α in the immunopathogenesis of rheumatoid arthritis. Arthritis Res Ther. 2017;19(1):261.
38. Li J, Ma L, Yu H, Yao Y, Xu Z, Lin W, et al. MicroRNAs as Potential Biomarkers for the Diagnosis of Chronic Kidney Disease: A Systematic Review and Meta-Analysis. Front Med (Lausanne). 2021;8:782561.
39. Wang B, Li Y, Hao X, Yang J, Han X, Li H, et al. Comparison of the Clinical Value of miRNAs and Conventional Biomarkers in AMI: A Systematic Review. Front Genet. 2021;12:668324.
40. Li SQ, Xie LY, Cai ZM, Wei HT, Xie MZ, Hu BL, et al. Systematic analyzing a five- miRNA panel and its diagnostic value of plasma expression in colorectal cancer. Mol Biol Rep. 2023;50(9):7253-61.
41. Garmaa G, Nagy R, Kói T, To UND, Gergő D, Kleiner D, et al. Panel miRNAs are potential diagnostic markers for chronic kidney diseases: a systematic review and meta-analysis. BMC Nephrol. 2024;25(1):261.
42. Kim DH, Park H, Choi YJ, Im K, Lee CW, Kim D-S, et al. Identification of exosomal microRNA panel as diagnostic and prognostic biomarker for small cell lung cancer. Biomarker Research. 2023;11(1):80.
43. Wen Z, Li Y, Zhao Z, Li R, Li X, Lu C, et al. A serum panel of three microRNAs may serve as possible biomarkers for kidney renal clear cell carcinoma. Cancer Cell International. 2024;24(1):18.
44. Pigati L, Yaddanapudi SC, Iyengar R, Kim DJ, Hearn SA, Danforth D, et al. Selective release of microRNA species from normal and malignant mammary epithelial cells. PLoS One. 2010;5(10):e13515.
45. Yang QY, Yang KP, Li ZZ. MiR-22 restrains proliferation of rheumatoid arthritis by targeting IL6R and may be concerned with the suppression of NF-κB pathway. Kaohsiung J Med Sci. 2020;36(1):20-6.
46. Zhang C, Fang L, Liu X, Nie T, Li R, Cui L, et al. miR-22 inhibits synovial fibroblasts proliferation and proinflammatory cytokine production in RASF via targeting SIRT1. Gene. 2020;724:144144.
47. Chang L, Zhou R. Histone methyltransferase EZH2 in proliferation, invasion, and migration of fibroblast-like synoviocytes in rheumatoid arthritis. J Bone Miner Metab. 2022;40(2):262-74.
48. Liu F, Wang Y, Huang D, Sun Y. LncRNA HOTAIR regulates the PI3K/AKT pathway via the miR-126-3p/PIK3R2 axis to participate in synovial angiogenesis in rheumatoid arthritis. Immun Inflamm Dis. 2023;11(10):e1064.
49. Liu W, Song J, Feng X, Yang H, Zhong W. LncRNA XIST is involved in rheumatoid arthritis fibroblast-like synoviocytes by sponging miR-126-3p via the NF-κB pathway. Autoimmunity. 2021;54(6):326-35.
2. Venetsanopoulou AI, Alamanos Y, Voulgari PV, Drosos AA. Epidemiology and Risk Factors for Rheumatoid Arthritis Development. Mediterr J Rheumatol. 2023;34(4):404-13.
3. Scherer HU, Häupl T, Burmester GR. The etiology of rheumatoid arthritis. Journal of Autoimmunity. 2020;110:102400.
4. Jang S, Kwon EJ, Lee JJ. Rheumatoid Arthritis: Pathogenic Roles of Diverse Immune Cells. Int J Mol Sci. 2022;23(2).
5. Cheng Q, Chen X, Wu H, Du Y. Three hematologic/immune system-specific expressed genes are considered as the potential biomarkers for the diagnosis of early rheumatoid arthritis through bioinformatics analysis. J Transl Med. 2021;19(1):18.
6. O'Brien J, Hayder H, Zayed Y, Peng C. Overview of MicroRNA Biogenesis, Mechanisms of Actions, and Circulation. Front Endocrinol (Lausanne). 2018;9:402.
7. Pozniak T, Shcharbin D, Bryszewska M. Circulating microRNAs in Medicine. Int J Mol Sci. 2022;23(7).
8. Wu Y, Li Q, Zhang R, Dai X, Chen W, Xing D. Circulating microRNAs: Biomarkers of disease. Clin Chim Acta. 2021;516:46-54.
9. Peng X, Wang Q, Li W, Ge G, Peng J, Xu Y, et al. Comprehensive overview of microRNA function in rheumatoid arthritis. Bone Res. 2023;11(1):8.
10. Kent WJ, Sugnet CW, Furey TS, Roskin KM, Pringle TH, Zahler AM, et al. The human genome browser at UCSC. Genome research. 2002;12(6):996-1006.
11. Nassar LR, Barber GP, Benet-Pagès A, Casper J, Clawson H, Diekhans M, et al. The UCSC Genome Browser database: 2023 update. Nucleic Acids Research. 2023;51(D1):D1188-D95.
12. Kozomara A, Birgaoanu M, Griffiths-Jones S. miRBase: from microRNA sequences to function. Nucleic Acids Res. 2019;47(D1):D155-d62.
13. Zuker M. Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Research. 2003;31(13):3406-15.
14. Ru Y, Kechris KJ, Tabakoff B, Hoffman P, Radcliffe RA, Bowler R, et al. The multiMiR R package and database: integration of microRNA-target interactions along with their disease and drug associations. Nucleic Acids Res. 2014;42(17):e133.
15. R Core Team (2022). R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. URL https://www.R-project.org/.
16. RStudio Team (2022). RStudio: Integrated Development Environment for R. RStudio, PBC, Boston, MA URL http://www.rstudio.com/.
17. Li J-H, Liu S, Zhou H, Qu L-H, Yang J-H. starBase v2.0: decoding miRNA-ceRNA, miRNA-ncRNA and protein–RNA interaction networks from large-scale CLIP-Seq data. Nucleic Acids Research. 2013;42(D1):D92-D7.
18. Shannon P, Markiel A, Ozier O, Baliga NS, Wang JT, Ramage D, et al. Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Res. 2003;13(11):2498-504.
19. Chin C-H, Chen S-H, Wu H-H, Ho C-W, Ko M-T, Lin C-Y. cytoHubba: identifying hub objects and sub-networks from complex interactome. BMC Systems Biology. 2014;8(4):S11.
20. Xie Z, Bailey A, Kuleshov MV, Clarke DJ, Evangelista JE, Jenkins SL, et al. Gene set knowledge discovery with enrichr. Current protocols. 2021;1(3):e90.
21. H. Wickham. ggplot2: Elegant Graphics for Data Analysis. Springer-Verlag New York, 2016.
22. Zamani M, Cassiman JJ. Reevaluation of the importance of polymorphic HLA class II alleles and amino acids in the susceptibility of individuals of different populations to type I diabetes. American journal of medical genetics. 1998;76(2):183-94.
23. Davatchi F, Sandoughi M, Moghimi N, Jamshidi AR, Tehrani Banihashemi A, Zakeri Z, et al. Epidemiology of rheumatic diseases in Iran from analysis of four COPCORD studies. International journal of rheumatic diseases. 2016;19(11):1056-62.
24. Ding Q, Hu W, Wang R, Yang Q, Zhu M, Li M, et al. Signaling pathways in rheumatoid arthritis: implications for targeted therapy. Signal Transduct Target Ther. 2023;8(1):68.
25. Kim JW, Suh CH. Systemic Manifestations and Complications in Patients with Rheumatoid Arthritis. J Clin Med. 2020;9(6).
26. Steiner G, Verschueren P, Van Hoovels L, Studenic P, Bossuyt X. Classification of rheumatoid arthritis: is it time to revise the criteria? RMD Open. 2024;10(2).
27. Metcalf GAD. MicroRNAs: circulating biomarkers for the early detection of imperceptible cancers via biosensor and machine-learning advances. Oncogene. 2024;43(28):2135-42.
28. Gayosso-Gómez LV, Ortiz-Quintero B. Circulating MicroRNAs in Blood and Other Body Fluids as Biomarkers for Diagnosis, Prognosis, and Therapy Response in Lung Cancer. Diagnostics (Basel). 2021;11(3).
29. Evangelatos G, Fragoulis GE, Koulouri V, Lambrou GI. MicroRNAs in rheumatoid arthritis: From pathogenesis to clinical impact. Autoimmun Rev. 2019;18(11):102391.
30. Tang J, Lin J, Yu Z, Jiang R, Xia J, Yang B, et al. Identification of circulating miR-22-3p and let-7a-5p as novel diagnostic biomarkers for rheumatoid arthritis. Clin Exp Rheumatol. 2022;40(1):69-77.
31. Cieśla M, Kolarz B, Majdan M, Darmochwał-Kolarz D. Plasma micro-RNA-22 is associated with disease activity in well-established rheumatoid arthritis. Clin Exp Rheumatol. 2022;40(5):945-51.
32. Ormseth MJ, Solus JF, Sheng Q, Ye F, Song H, Wu Q, et al. The Endogenous Plasma Small RNAome of Rheumatoid Arthritis. ACR Open Rheumatol. 2020;2(2):97-105.
33. Renman E, Brink M, Ärlestig L, Rantapää-Dahlqvist S, Lejon K. Dysregulated microRNA expression in rheumatoid arthritis families-a comparison between rheumatoid arthritis patients, their first-degree relatives, and healthy controls. Clin Rheumatol. 2021;40(6):2387-94.
34. Cunningham CC, Wade S, Floudas A, Orr C, McGarry T, Wade S, et al. Serum miRNA Signature in Rheumatoid Arthritis and "At-Risk Individuals". Front Immunol. 2021;12:633201.
35. Ormseth MJ, Solus JF, Sheng Q, Chen SC, Ye F, Wu Q, et al. Plasma miRNAs improve the prediction of coronary atherosclerosis in patients with rheumatoid arthritis. Clin Rheumatol. 2021;40(6):2211-9.
36. Abdelaleem OO, Fouad NA, Shaker OG, Ahmed TI, Abdelghaffar NK, Eid HM, et al. Serum miR-224, miR-760, miR-483-5p, miR-378 and miR-375 as potential novel biomarkers in rheumatoid arthritis. Int J Clin Pract. 2021;75(11):e14651.
37. Lai NS, Yu HC, Tung CH, Huang KY, Huang HB, Lu MC. The role of aberrant expression of T cell miRNAs affected by TNF-α in the immunopathogenesis of rheumatoid arthritis. Arthritis Res Ther. 2017;19(1):261.
38. Li J, Ma L, Yu H, Yao Y, Xu Z, Lin W, et al. MicroRNAs as Potential Biomarkers for the Diagnosis of Chronic Kidney Disease: A Systematic Review and Meta-Analysis. Front Med (Lausanne). 2021;8:782561.
39. Wang B, Li Y, Hao X, Yang J, Han X, Li H, et al. Comparison of the Clinical Value of miRNAs and Conventional Biomarkers in AMI: A Systematic Review. Front Genet. 2021;12:668324.
40. Li SQ, Xie LY, Cai ZM, Wei HT, Xie MZ, Hu BL, et al. Systematic analyzing a five- miRNA panel and its diagnostic value of plasma expression in colorectal cancer. Mol Biol Rep. 2023;50(9):7253-61.
41. Garmaa G, Nagy R, Kói T, To UND, Gergő D, Kleiner D, et al. Panel miRNAs are potential diagnostic markers for chronic kidney diseases: a systematic review and meta-analysis. BMC Nephrol. 2024;25(1):261.
42. Kim DH, Park H, Choi YJ, Im K, Lee CW, Kim D-S, et al. Identification of exosomal microRNA panel as diagnostic and prognostic biomarker for small cell lung cancer. Biomarker Research. 2023;11(1):80.
43. Wen Z, Li Y, Zhao Z, Li R, Li X, Lu C, et al. A serum panel of three microRNAs may serve as possible biomarkers for kidney renal clear cell carcinoma. Cancer Cell International. 2024;24(1):18.
44. Pigati L, Yaddanapudi SC, Iyengar R, Kim DJ, Hearn SA, Danforth D, et al. Selective release of microRNA species from normal and malignant mammary epithelial cells. PLoS One. 2010;5(10):e13515.
45. Yang QY, Yang KP, Li ZZ. MiR-22 restrains proliferation of rheumatoid arthritis by targeting IL6R and may be concerned with the suppression of NF-κB pathway. Kaohsiung J Med Sci. 2020;36(1):20-6.
46. Zhang C, Fang L, Liu X, Nie T, Li R, Cui L, et al. miR-22 inhibits synovial fibroblasts proliferation and proinflammatory cytokine production in RASF via targeting SIRT1. Gene. 2020;724:144144.
47. Chang L, Zhou R. Histone methyltransferase EZH2 in proliferation, invasion, and migration of fibroblast-like synoviocytes in rheumatoid arthritis. J Bone Miner Metab. 2022;40(2):262-74.
48. Liu F, Wang Y, Huang D, Sun Y. LncRNA HOTAIR regulates the PI3K/AKT pathway via the miR-126-3p/PIK3R2 axis to participate in synovial angiogenesis in rheumatoid arthritis. Immun Inflamm Dis. 2023;11(10):e1064.
49. Liu W, Song J, Feng X, Yang H, Zhong W. LncRNA XIST is involved in rheumatoid arthritis fibroblast-like synoviocytes by sponging miR-126-3p via the NF-κB pathway. Autoimmunity. 2021;54(6):326-35.
| Files | ||
| Issue | Articles in Press | |
| Section | Original Article(s) | |
| Keywords | ||
| Biomarker MicroRNA miR-126-3p miR-22-3p miR-760 Rheumatoid arthritis | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
Raghibi A, Mohajeri Khorasani A, Damavandi E, Rostamian A, Abjoushak Mahmoudabad M, Choobineh H, Ghadami M, Kabuli M. Integrated Analysis of Circulating miR-22-3p, miR-126-3p, and miR-760 as Non-invasive Biomarkers for Rheumatoid Arthritis. Iran J Allergy Asthma Immunol. 2026;:1-20.
