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 4

LERFS functions by interacting with hnRNP Q.

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LERFS functions by interacting with hnRNP Q. (A) Experimental design for...
(A) Experimental design for pulldown assays and identification of LERFS-associated cellular proteins. LERFS RNA was biotinylated by in vitro transcription, refolded, and incubated with lysates of RA FLSs. (B) Silver staining of biotinylated LERFS-associated proteins. A LERFS-specific band was excised and analyzed by MS, which identified hnRNP Q. (C) Western blot of proteins from LERFS-pulldown assays. (D) RIP evaluation of the interaction between hnRNP Q and LERFS within RA FLSs using an anti–hnRNP Q antibody (5 μg), with IgG (5 μg) as a NC. SnRNP70 was used as a positive control (right). **P < 0.01 and ***P < 0.001 versus IgG, by Student’s t test.U1, U1 small nuclear RNA (snRNA). (E) Comparison of LERFS binding with hnRNP Q between RA FLSs and HC FLSs. Data are shown as the mean ± SEM of 3 independent experiments involving 3 different RA patients and HCs. ***P < 0.001 versus HC FLSs, by Student’s t test. (F–H) Effect of hnRNP Q knockdown on the migration, invasion, and proliferation of RA FLSs. Representative images are shown (original magnification, ×200). Data for relative migration (F), invasion (G), and proliferation (H) are shown as the mean ± SEM of 5 independent experiments involving 5 different RA patients. *P < 0.05, **P < 0.01, and *** P < 0.001 versus siControl (siC) or vector, by Student’s t test. (I–K) Overexpression of hnRNP Q suppressed the migration (I), invasion (J), and proliferation (K) of RA FLSs. Representative images are shown. Original magnification, ×100 (I and J); ×200 (K). Data in I–K were normalized to the control group (vector) and are presented as the mean ± SEM of 5 independent experiments involving 5 different RA patients. **P < 0.01 and *** P < 0.001 versus siC or vector, by Student’s t test.