Serum-derived exosomes containing NEAT1 promote the occurrence of rheumatoid arthritis through regulation of miR-144-3p/ROCK2 axis

Abstract

Background: Evidence has demonstrated that non-coding RNAs (ncRNAs) could be delivered efficiently to recipient cells using exosomes as a carrier. Additionally, long ncRNA nuclear enriched abundant transcript 1 (NEAT1) is emerging as a vital regulatory molecule in the progression of rheumatoid arthritis (RA). The aim of this study was to identify the NEAT1/miR-144-3p/Rho-associated protein kinase 2 (ROCK2) functional network regulating the WNT signaling pathway in RA. Methods: In vivo, a collagen-induced arthritis (CIA) model was established to analyze the effects of blood exosomes on the incidence, clinical score, and bone degradation of RA. In vitro, the CD4+T cells were characterized by flow cytometry and the cell activities were analyzed in the presence of exosome treatment alone or in combination with altered expression of NEAT1, miR-144-3p or Rho-associated protein kinase 2 (ROCK2). The expression of NEAT1, miR-144-3p, ROCK2, and corresponding proteins in the WNT signaling pathway was detected by RT-qPCR and western blot techniques. The binding profile of NEAT1 to miR-144-3p was evaluated via a combination approach of luciferase activity assay, RNA immunoprecipitation, and RNA pull-down experiments. Results: Blood exosomes extracted from RA patients increased the incidence of RA and bone destruction significantly. Overexpression of NEAT1 or ROCK2 promoted immune cell (CD4+T cells) proliferation, Th17 cell differentiation, and cell migration in response to stimulus, whereas knockout of the NEAT1 gene induced the expression of miR-144-3p in CD4+T cells. ROCK2 exogenous expression inhibited the expression of miR-144-3p, inducing activation of the WNT signaling pathway. Conclusion: A novel regulatory pathway NEAT1/miR-144-3p/ROCK2/WNT in RA was investigated as a potential target for RA therapy.

Long non‐coding RNA NEAT1 and its targets (microRNA‐21 and microRNA‐125a) in rheumatoid arthritis: Altered expression and potential to monitor disease activity and treatment outcome

“…The possible reasons might be that: (a) elevated lnc‐NEAT1 and decreased miR‐21 and miR‐125a could promote inflammatory cytokines and immune response, which consequently resulted in elevated inflammation in RA patients 7 , 19 , 20 , 23 ; and (b) increased lnc‐NEAT1 and declined miR‐21 and miR‐125a might inhibit new bone formation and accelerate bone degradation through regulating several signaling pathways (such as Janus kinase 2/signal transducer, activator of transcription‐3, and E26 transformation‐specific‐1 pathways), which could lead to high disease activity in RA patients. 24 , 25 , 26 Therefore, elevated lnc‐NEAT1 but declined miR‐21 and miR‐125a were correlated with inflammation and disease activity in RA.…”

Role of Exosomal Non-Coding RNAs in Bone-Related Diseases

“…That study provides new ideas for the osteogenic effects of exosomes in BMSCs carrying specific tsRNAs on SONFH. Besides, there is evidence that exosomal lncRNA nuclear enriched abundant transcript (NEAT)1 isolated from the serum of patients with rheumatoid arthritis can promote the development of rheumatoid arthritis through the miR-144-3p/ROCK2 axis ( Liu et al, 2021 ). This NEAT1/ miR-144–3p/ROCK2 regulatory pathway may become a new treatment target for rheumatoid arthritis.…”

LncRNA Expression Profiles in Systemic Lupus Erythematosus and Rheumatoid Arthritis: Emerging Biomarkers and Therapeutic Targets

“…Subsequent studies have also shown that, compared with the exosomes from normal controls (n=20), there was a significant increase in the expression of NEAT1 in the exosomes of patients with RA (n=68). Also, NEAT1 might act as a ceRNA for miR144-3p to restrict its function, and thus increase the expression of the miR144-3p-targeted gene (Rho associated coiled-coil containing protein kinase 2, ROCK2) in CD4 + T cells, promoting the progression of RA ( 129 ). Another study showed that the levels of a set of lncRNAs, HOTAIR, Luca-15 Specific Transcript (LUST), anti-NOS2A, MEG, TUG1, NEAT1, Small Nucleolar RNA Host Gene 4 (SNHG4), Highly Accelerated Region 1B (HAR1B), and GAS5, have higher expression levels in seral exosomes of patients with RA (n=28) than in the seral exosomes of normal controls (n=10) ( 89 ).…”