Study of The Correlation between miR-106a, miR-125b, and miR-330 on Multiple Sclerosis Patients by Targeting TNFSF4 and SP1 in NF-кb/TNF-α Pathway: A Case-Control Study

Document Type : Original Article


1 Medical Biotechnology Research Center, Ashkezar Branch, Islamic Azad University, Ashkezar, Yazd, Iran

2 Medical Genetics Research Center of Genome, Isfahan University of Medical Sciences, Isfahan, Iran

3 Cellular, Molecular, and Genetics Research Center, Isfahan University of Medical Sciences, Isfahan, Iran

4 Department of Biology, Faculty of Science, Shahid Chamran University of Ahvaz, Ahvaz, Iran

5 Department of Neurology, Isfahan Neurosciences Research Center, Isfahan University of Medical Sciences, Isfahan, Iran

6 Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran


Objective: Multiple sclerosis (MS) is a complex multifactorial neuro-inflammatory disorder. This complexity arises from the evidence suggesting that MS is developed by interacting with environmental and genetic factors. This study aimed to evaluate the miR-106a, miR-125b, and miR330- expression levels in relapsing-remitting multiple sclerosis (RRMS) patients. The miRNAs' impact on TNFSF4 and Sp1 genes through the NF-кB/TNF-α signaling pathway was analyzed by measuring the expression levels in case and controls.
Materials and Methods: In this in silico-experimental study, we evaluated the association of miR-106a, miR- 125b, and miR330- with TNFSF4 and SP1 gene expression levels in 60 RRMS patients and 30 healthy controls by real-time polymerase chain reaction (PCR).
Results: The expression levels of miR-330, miR-106a, and miR125-b in blood samples of RRMS patients were predominantly reduced. The expression of TNFSF4 in patients demonstrated a significant enhancement, in contrast to the diminishing Sp1 gene expression level in controls.
Conclusion: Our findings indicated an association between miR-106a and miR-330 and miR125-b expression and RRMS in our study population. Our data suggested that the miR106-a, miR125-b, and mir330- expression are correlated with TNFSF4 and Sp1 gene expression levels.


1. Lassmann H. Multiple sclerosis pathology. Cold Spring Harb Perspect Med. 2018; 8(3): a028936.
2. Montalban X, Hauser SL, Kappos L, Arnold DL, Bar-Or A, Comi G, et al. Ocrelizumab versus placebo in primary progressive multiple sclerosis. N Engl J Med. 2017; 376(3): 209-220.
3. Goldenberg MM. Multiple sclerosis review. P T. 2012; 37(3): 175-84.
4. Stangel M, Penner IK, Kallmann BA, Lukas C, Kieseier BC. Towards the implementation of ‘no evidence of disease activity’ in multiple sclerosis treatment: the multiple sclerosis decision model. Ther Adv Neurol Disord. 2015; 8(1): 3-13.
5. Patsopoulos NA. Genetics of multiple sclerosis: an overview and new directions. Cold Spring Harb Perspect Med. 2018; 8(7): a028951.
6. Kahl KG, Kruse N, Faller H, Weiss H, Rieckmann P. Expression of tumor necrosis factor-alpha and interferon-gamma mRNA in blood cells correlates with depression scores during an acute attack in patients with multiple sclerosis. Psychoneuroendocrinology. 2002; 27(6): 671-681.
7. Ribeiro CM, Oliveira SR, Alfieri DF, Flauzino T, Kaimen-Maciel DR, Simão ANC, et al. Tumor necrosis factor alpha (TNF-α) and its soluble receptors are associated with disability, disability progression and clinical forms of multiple sclerosis. Inflamm Res. 2019; 68(12): 1049-1059.
8. Fiedler SE, George JD, Love HN, Kim E, Spain R, Bourdette D, et al. Analysis of IL-6, IL-1β and TNF-α production in monocytes isolated from multiple sclerosis patients treated with disease modifying drugs. J Syst Integr Neurosci. 2017; 3(3): 10.
9. Yang G, Parkhurst CN, Hayes S, Gan WB. Peripheral elevation of TNF-α leads to early synaptic abnormalities in the mouse somatosensory cortex in experimental autoimmune encephalomyelitis. Proc Natl Acad Sci USA. 2013; 110(25): 10306-10311.
10. Dong Y, Dekens DW, De Deyn PP, NaudéPJW, Eisel ULM. Targeting of tumor necrosis factor alpha receptors as a therapeutic strategy for neurodegenerative disorders. Antibodies. 2015; 4(4): 369-408.
11. An J, Ding S, Hu X, Sun L, Gu Y, Xu Y, et al. Preparation, characterization and application of anti-human OX40 ligand (OX40L) monoclonal antibodies and establishment of a sandwich ELISA for autoimmune diseases detection. Int Immunopharmacol. 2019; 67: 260-267.
12. van Wanrooij EJ, van Puijvelde GH, de Vos P, Yagita H, van Berkel TJ, Kuiper J. Interruption of the Tnfrsf4/Tnfsf4 (OX40/OX40L) pathway attenuates atherogenesis in low-density lipoprotein receptor-deficient mice. Arterioscler Thromb Vasc Biol. 2007; 27(1): 204-210.
13. Takahashi Y, Tanaka R, Yamamoto N, Tanaka Y. Enhancement of OX40-induced apoptosis by TNF coactivation in OX40-expressing T cell lines in vitro leading to decreased targets for HIV type 1 production. AIDS Res Hum Retroviruses. 2008; 24(3): 423-435.
14. Ping D, Boekhoudt G, Zhang F, Morris A, Philipsen S, Warren ST, et al. Sp1 binding is critical for promoter assembly and activation of the MCP-1 gene by tumor necrosis factor. J Biol Chem. 2000; 275(3): 1708-1714.
15. Hirano F, Tanaka H, Hirano Y, Hiramoto M, Handa H, Makino I, et al. Functional interference of Sp1 and NF-kappaB through the same DNA binding site. Mol Cell Biol. 1998; 18(3): 1266-1274.
16. Shende VR, Kim SM, Neuendorff N, Earnest DJ. MicroRNAs function as cis- and trans-acting modulators of peripheral circadian clocks. FEBS Lett. 2014; 588(17): 3015-3022.
17. Tufekci KU, Oner MG, Genc S, Genc K. MicroRNAs and multiple sclerosis. Autoimmune Diseases. 2011; 807426.
18. Honardoost MA, Naghavian R, Ahmadinejad F, Hosseini A, Ghaedi K. Integrative computational mRNA-miRNA interaction analyses of the autoimmune-deregulated miRNAs and wellknown Th17 differentiation regulators: An attempt to discover new potential miRNAs involved in Th17 differentiation. Gene. 2015; 572(2): 153-162.
19. Sanctuary MR, Huang RH, Jones AA, Luck ME, Aherne CM, Jedlicka P, et al. miR-106a deficiency attenuates inflammation in murine IBD models. Mucosal Immunol. 2019; 12(1): 200-211.
20. Kim BK, Yoo HI, Choi K, Yoon SK. miR-330-5p inhibits proliferation and migration of keratinocytes by targeting Pdia3 expression. FEBS J. 2015; 282(24): 4692-4702.
21. Mao Y, Chen H, Lin Y, Xu X, Hu Z, Zhu Y, et al. microRNA-330 inhibits cell motility by downregulating Sp1 in prostate cancer cells. Oncol Rep. 2013; 30(1): 327-333.
22. Sticht C, De La Torre C, Parveen A, Gretz N. miRWalk: An online resource for prediction of microRNA binding sites. PLoS One. 2018; 13(10): e0206239.
23. Piñero J, Ramírez-Anguita JM, Saüch-Pitarch J, Ronzano F, Centeno E, Sanz F, et al. The DisGeNET knowledge platform
for disease genomics: 2019 update. Nucleic Acids Res. 2020; 48(D1): D845-D855.
24. Jassal B, Matthews L, Viteri G, Gong C, Lorente P, Fabregat A, et al. The reactome pathway knowledgebase. Nucleic Acids Res. 2020; 48(D1): D498-D503.
25. 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): 2498-2504.
26. Lu Q. The critical importance of epigenetics in autoimmunity. J Autoimmun. 2013; 41: 1-5.
27. Fathullahzadeh S, Mirzaei H, Honardoost MA, Sahebkar A, Salehi M. Circulating microRNA-192 as a diagnostic biomarker in human chronic lymphocytic leukemia. Cancer Gene Ther. 2016; 23(10): 327-332.
28. Hauser SL, Bar-Or A, Comi G, Giovannoni G, Hartung HP, Hemmer B, et al. Ocrelizumab versus interferon beta-1a in relapsing multiple sclerosis. N Engl J Med. 2017; 376(3): 221-234.
29. Regev K, Healy BC, Khalid F, Paul A, Chu R, Tauhid S, et al. Association between serum micrornas and magnetic resonance imaging measures of multiple sclerosis severity. JAMA Neurol. 2017; 74(3): 275-285.
30. Choi I, Woo JH, Jou I, Joe EH. PINK1 deficiency decreases expression levels of mir-326, mir-330, and mir-3099 during brain development and neural stem cell differentiation. Exp Neurobiol. 2016; 25(1): 14-23.
31. Cox MB, Cairns MJ, Gandhi KS, Carroll AP, Moscovis S, Stewart GJ, et al. MicroRNAs miR-17 and miR-20a inhibit T cell
activation genes and are under-expressed in MS whole blood. PLoS One. 2010; 5(8): e12132.
32. Tan L, Yu JT, Liu QY, Tan MS, Zhang W, Hu N, et al. Circulating miR-125b as a biomarker of Alzheimer’s disease. J Neurol Sci. 2014; 336(1-2): 52-56.
33. Sicotte NL, Voskuhl RR. Onset of multiple sclerosis associated with anti-TNF therapy. Neurology. 2001; 57(10): 1885-1888.
34. Sonar S, Lal G. Role of tumor necrosis factor superfamily in neuroinflammation and autoimmunity. Front Immunol.  2015; 6: 364.
35. Kemanetzoglou E, Andreadou E. CNS demyelination with TNF-α blockers. Curr Neurol Neurosci Rep. 2017; 17(4): 36.