- Poster presentation
- Open Access
Association of microRNA-221/222 and -323-3p with rheumatoid arthritis via predictions using the human TNF transgenic mouse model
Arthritis Research & Therapyvolume 14, Article number: P59 (2012)
MicroRNAs (miRs), a class of small non-coding RNA molecules, act as posttranscriptional regulators and are involved in a plethora of cellular functions. miRs have attracted a great deal of attention as potential therapeutic targets, as the sequence-specific mode in which they act, allows the simultaneous targeting of multiple target genes, often members of the same biological pathway(s) . Previous studies have demonstrated that miRs are dysregulated and functionally involved in rheumatoid arthritis (RA) [2–9]. In this study we sought to identify novel miR associations in synovial fibroblasts (SFs), a key pathogenic cell type in RA [10, 11], by performing miR expression profiling on cells isolated from the human TNF transgenic mouse model (TghuTNF, Tg197)  and patients biopsies.
Materials and methods
miR expression in SFs from TghuTNF and WT control mice were determined by deep sequencing and the arthritic profile was established by pairwise comparisons. qRT-PCR analysis was utilised for profile validation, miR and gene quantitation in patient SFs. Dysregulated miR target genes and pathways were predicted via bioinformatic algorithms.
Deep sequencing demonstrated that TghuTNF-SFs exhibit a distinct pathogenic profile with 22 significantly upregulated and 30 significantly downregulated miRs (fold change>1.5, p-value<0.05). qRT-PCR validation assays confirmed the dysregulation of miR-223, miR-146a and miR-155 previously associated with human RA pathology, as well as that of miR-221/222 and miR-323-3p. Notably, the latter were also found significantly upregulated in patient RASFs, suggesting their association with human RA pathology. Bioinformatic analysis suggested Wnt/Cadherin signaling as the most significant pathway targets of miR-221/222 and miR-323-3p and CSNK1A1 and BTRC, the negative regulators of β-catenin, amongst predicted gene targets. qRT-PCR assays confirmed the downregulation of these genes in RASFs, validating our hypothesis that the newly identified miRs may function to modulate Wnt/Cadherin signaling.
In this study, by performing comparative analyses between an established mouse model of arthritis and RA patient biopsies, we identified novel dysregulated miRs in RASFs potentially involved in pathways important for the pathogenic phenotype of these cells and highlighting the value of such cross-species comparative approaches .
Montgomery RL, van Rooij E: microRNA Regulation as a Therapeutic Strategy for Cardiovascular Disease. Curr Drug Targets. 2010, 11 (8): 936-942. 10.2174/138945010791591368.
Murata K, Yoshitomi H, Tanida S, Ishikawa M, Nishitani K, Ito H, Nakamura T: Plasma and synovial fluid microRNAs as potential biomarkers of rheumatoid arthritis and osteoarthritis. Arthritis Res Ther. 2010, 12 (3): R86-10.1186/ar3013.
Pauley KM, Satoh M, Chan AL, Bubb MR, Reeves WH, Chan EK: Upregulated miR-146a expression in peripheral blood mononuclear cells from rheumatoid arthritis patients. Arthritis Res Ther. 2008, 10 (4): R101-
Kawano S, Nakamachi Y: miR-124a as a key regulator of proliferation and MCP-1 secretion in synoviocytes from patients with rheumatoid arthritis. Ann Rheum Dis. 2011, 70: I88-I91. 10.1136/ard.2010.138669.
Stanczyk J, Ospelt C, Karouzakis E, Filer A, Raza K, Kolling C, Gay R, Buckley CD, Tak PP, Gay S, et al: Altered expression of MicroRNA-203 in rheumatoid arthritis synovial fibroblasts and its role in fibroblast activation. Arthritis Rheum. 2011, 63 (2): 373-381. 10.1002/art.30115.
Stanczyk J, Pedrioli DM, Brentano F, Sanchez-Pernaute O, Kolling C, Gay RE, Detmar M, Gay S, Kyburz D: Altered expression of MicroRNA in synovial fibroblasts and synovial tissue in rheumatoid arthritis. Arthritis Rheum. 2008, 58 (4): 1001-1009. 10.1002/art.23386.
Bluml S, Bonelli M, Niederreiter B, Puchner A, Mayr G, Hayer S, Koenders MI, van den Berg WB, Smolen J, Redlich K: Essential role for micro-RNA 155 in the pathogenesis of autoimmune arthritis. Arthritis Rheum. 2011, 63 (5): 1281-8. 10.1002/art.30281.
Kurowska-Stolarska M, Alivernini S, Ballantine LE, Asquith DL, Millar NL, Gilchrist DS, Reilly J, Ierna M, Fraser AR, Stolarski B, et al: MicroRNA-155 as a proinflammatory regulator in clinical and experimental arthritis. Proc Natl Acad Sci USA. 2011, 108 (27): 11193-8. 10.1073/pnas.1019536108.
Nakasa T, Shibuya H, Nagata Y, Niimoto T, Ochi M: The inhibitory effect of microRNA-146 expression on bone destruction in arthritis. Arthritis Rheum. 2011, 63 (6): 1582-90. 10.1002/art.30321.
Armaka M, Apostolaki M, Jacques P, Kontoyiannis DL, Elewaut D, Kollias G: Mesenchymal cell targeting by TNF as a common pathogenic principle in chronic inflammatory joint and intestinal diseases. J Exp Med. 2008, 205 (2): 331-337. 10.1084/jem.20070906.
Lefevre S, Knedla A, Tennie C, Kampmann A, Wunrau C, Dinser R, Korb A, Schnaker E-M, Tarner IH, Robbins PD, et al: Synovial fibroblasts spread rheumatoid arthritis to unaffected joints. Nat Med. 2009, 15 (12): 1414-1420. 10.1038/nm.2050.
Keffer J, Probert L, Cazlaris H, Georgopoulos S, Kaslaris E, Kioussis D, Kollias G: Transgenic mice expressing human tumour necrosis factor: a predictive genetic model of arthritis. EMBO J. 1991, 10 (13): 4025-4031.
Kollias G, Papadaki P, Apparailly F, Vervoordeldonk MJ, Holmdahl R, Baumans V, Desaintes C, Di Santo J, Distler J, Garside P, et al: Animal models for arthritis: innovative tools for prevention and treatment. Ann Rheum Dis. 2011, 70 (8): 1357-1362. 10.1136/ard.2010.148551.
This project was funded by the Masterswitch Project (HEALTH-F2-2008-223404), EURO-RA RTN (HPRN-CT-2002-00255) and IMI BtCure (grant agreement No 115142) grants to GK and SG. JR was supported by the Wellcome Trust grant 075491/Z/04. In SG also received funding from IAR-EPALINGES.