Long noncoding RNA LERFS negatively regulates rheumatoid synovial aggression and proliferation
Yaoyao Zou, … , Song Guo Zheng, Hanshi Xu
Yaoyao Zou, … , Song Guo Zheng, Hanshi Xu
Published September 10, 2018
Citation Information: J Clin Invest. 2018;128(10):4510-4524. https://doi.org/10.1172/JCI97965.
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Research Article Immunology

Long noncoding RNA LERFS negatively regulates rheumatoid synovial aggression and proliferation

Abstract

Fibroblast-like synoviocytes (FLSs) are critical to synovial aggression and joint destruction in rheumatoid arthritis (RA). The role of long noncoding RNAs (lncRNAs) in RA is largely unknown. Here, we identified a lncRNA, LERFS (lowly expressed in rheumatoid fibroblast-like synoviocytes), that negatively regulates the migration, invasion, and proliferation of FLSs through interaction with heterogeneous nuclear ribonucleoprotein Q (hnRNP Q). Under healthy conditions, by binding to the mRNA of RhoA, Rac1, and CDC42 — the small GTPase proteins that control the motility and proliferation of FLSs — the LERFS–hnRNP Q complex decreased the stability or translation of target mRNAs and downregulated their protein levels. But in RA FLSs, decreased LERFS levels induced a reduction of the LERFS–hnRNP Q complex, which reduced the binding of hnRNP Q to target mRNA and therefore increased the stability or translation of target mRNA. These findings suggest that a decrease in synovial LERFS may contribute to synovial aggression and joint destruction in RA and that targeting the lncRNA LERFS may have therapeutic potential in patients with RA.

Authors

Yaoyao Zou, Siqi Xu, Youjun Xiao, Qian Qiu, Maohua Shi, Jingnan Wang, Liuqin Liang, Zhongping Zhan, Xiuyan Yang, Nancy Olsen, Song Guo Zheng, Hanshi Xu

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Figure 1

Decreased levels of LERFS lncRNA in FLSs and STs from patients with RA.

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Decreased levels of LERFS lncRNA in FLSs and STs from patients with RA. ...
(A) Total RNA harvested from RA FLSs (n = 3) and HC FLSs (n = 3) was screened by microarray analysis. Microarray heatmap of differentially expressed lncRNAs. (B) Volcano plot shows differentially expressed lncRNAs between RA FLSs and HC FLSs. P < 0.05, by Student’s t test. (C) The expression level of LERFS was validated by RT-qPCR in HC FLSs (n = 29) and RA FLSs (n = 34). Ct values were normalized to GAPDH. Data are presented as the mean ± SEM. **P < 0.01 versus HCs, by Student’s t test. (D) RA FLSs were stimulated with IL-1β (10 ng/ml), TNF-α (10 ng/ml), PDGF-BB (10 ng/ml), IL-17 (10 ng/ml), LPS (10 ng/ml), synovial fluid (SF), or synovial fluid containing IgG (SF + IgG) or a PDGF-neutralizing antibody (SF + anti-PDGF) (50 ng/ml) for 24 hours. (E and F) RA FLSs were treated with MTX (E) or DXM (F) for 24 hours. (D–F) *P < 0.05, **P < 0.01, and ***P < 0.001 versus untreated control, by 1-way ANOVA with Bonferroni’s post hoc comparison (n = 5). (G) Localization of LERFS was evaluated by RNA FISH assay. For silencing of LERFS, HC FLSs were transfected with a specific mixture of siRNA and ASO for LERFS (siLERFS). Shown are representative images of LERFS (green) and nuclei (blue). Graph shows the quantification of staining intensity for 5 different RA patients and HCs. Original magnification, ×630. **P < 0.01 versus HCs, by Student’s t test. (H) LERFS expression, detected by ISH staining, on STs from HCs and RA patients. Shown are representative images and quantification of the percentage of LERFS-positive cells for 5 different RA patients or HCs. Also shown is a representative image of RA in remission from 2 remitted patients treated with MTX and TNF-α inhibitor. A scrambled probe was used as a NC. Red arrows indicate LERFS-positive (blue) cells. Original magnification, ×400. ***P < 0.001 versus HCs, by Student’s t test. C, control. Data are presented as the mean ± SEM.