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Downregulation of lncRNA ITSN1-2 correlates with decreased disease risk and activity of rheumatoid arthritis (RA), and reduces RA fibroblast-like synoviocytes proliferation and inflammation via inhibiting NOD2/RIP2 signaling pathway.

ABSTRACT: This study aimed to investigate the effect of lnc-ITSN1-2 knockdown on cell proliferation, apoptosis, inflammation and mRNA expression patterns in rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS), and the correlation of its synovium tissue expression with disease risk, inflammatory cytokines and disease activity of RA. Control shRNA plasmids and lnc-ITSN1-2 shRNA plasmids were transfected into RA FLS, and then cell proliferation, apoptosis, inflammatory cytokines expressions were evaluated. Subsequently, mRNA sequencing and bioinformatics analyses were conducted, and rescue experiment of nucleotide-binding oligomerization domain 2 (NOD2) mRNA overexpression on alleviating the functions of lnc-ITSN1-2 was performed. Additionally, lnc-ITSN1-2 and NOD2 mRNA expressions in synovial tissue in 30 RA patients and 15 controls were measured. Lnc-ITSN1-2 expression was increased in RA FLS compared with normal FLS. Lnc-ITSN1-2 knockdown inhibited RA FLS proliferation and inflammation while promoted RA FLS apoptosis. mRNA sequencing and bioinformatics analyses revealed 144 upregulated and 98 downregulated genes by lnc-ITSN1-2 knockdown, which were enriched in regulating inflammatory responses and cytokines related pathways. NOD2 was selected for rescue experiment, which disclosed that upregulating NOD2 alleviated the effect of lnc-ITSN1-2 knockdown on cell proliferation, apoptosis and inflammation in RA FLS. In addition, synovial tissue lnc-ITSN1-2 positively associated with NOD2 mRNA, and both of them positively correlated with disease risk, inflammation and activity of RA. Downregulation of lnc-ITSN1-2 correlates with decreased disease risk and activity of RA, and reduces RA FLS proliferation and inflammation via regulating NOD2/RIP2 signaling pathway.

SUBMITTER: Yue T

PROVIDER: S-EPMC6731396 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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Downregulation of lncRNA ITSN1-2 correlates with decreased disease risk and activity of rheumatoid arthritis (RA), and reduces RA fibroblast-like synoviocytes proliferation and inflammation via inhibiting NOD2/RIP2 signaling pathway.

Yue Tao T Fan Xiaolei X Zhang Zhanming Z Liu Zhaoyi Z Guo Mengru M Bai Fengmin F Gong Xumin X Gao Chenxin C Xiao Lianbo L

American journal of translational research 20190815 8

This study aimed to investigate the effect of lnc-ITSN1-2 knockdown on cell proliferation, apoptosis, inflammation and mRNA expression patterns in rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS), and the correlation of its synovium tissue expression with disease risk, inflammatory cytokines and disease activity of RA. Control shRNA plasmids and lnc-ITSN1-2 shRNA plasmids were transfected into RA FLS, and then cell proliferation, apoptosis, inflammatory cytokines expressions were eva ...[more]

PMID: 31497189

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Project description:BackgroundFibroblast-like synoviocytes (FLSs), which can migrate and directly invade the cartilage and the bone, are crucial players in joint damage in rheumatoid arthritis (RA). Nevertheless, the detailed mechanisms underlying the aberrant activation of RA FLSs remain unclear. Several studies have attempted to explore the relationship between long non-coding RNAs (lncRNAs) and RA pathology; however, the role of lncRNAs in RA is unknown. The present study aimed to determine the functions of tumor necrosis factor-α and heterogeneous nuclear ribonucleoprotein L-related immunoregulatory lincRNA (THRIL) in RA FLSs migration and invasion.MethodsSmall interfering RNA targeting THRIL or lentivirus overexpressing THRIL was used to knockdown or overexpress THRIL. Quantitative reverse transcription polymerase chain reaction (PCR) was employed for the detection of RNA expression. The proliferation rate of RA FLSs was measured using a 5-ethynyl-2'-deoxyuridine (EdU) incorporation assay. Migration and invasion were detected using a transwell chamber. Downstream targets were identified using a human cell cycle real-time PCR array and a human cell motility real-time PCR array.ResultsA significant decrease in THRIL expression was found in RA FLSs compared with cells from healthy control (HC)patients. THRIL is mainly localized in the nucleus. Knockdown of THRIL increased the proliferation, migration, and invasion of RA FLSs. In contrast, THRIL overexpression had the opposite effect. THRIL knockdown increased interleukin-1β (IL-1β)-triggered expression of matrix metalloproteinase (MMP)-1, MMP-3, and MMP-13. THRIL overexpression led to a significant decrease in MMP-13 expression in response to stimulation with IL-1β. Furthermore, we observed that the expression levels of cyclin-dependent kinase 1 (CDK1) and G2 and S phase-expressed-1 (GTSE1), both of which are associated with cellular mobility and proliferation, were downregulated with THRIL overexpression.ConclusionsReduced expression of lncRNA THRIL represses the proliferation, migration, and invasion of RA FLSs, suggesting that lncRNA THRIL might be a potential target for RA therapy.

Project description:ObjectiveUp-regulation of glucose metabolism has been implicated not only in tumor cell growth but also in immune cells upon activation. However, little is known about the metabolite profile in rheumatoid arthritis (RA), particularly in fibroblast-like synoviocytes (FLS). This study was undertaken to evaluate whether changes in glucose metabolism in RA FLS could play a role in inflammation and joint damage.MethodsSynovium and FLS were obtained from patients with RA and patients with osteoarthritis (OA). The rate of glycolysis after stimulation of FLS with lipopolysaccharide and platelet-derived growth factor BB was measured using glycolysis stress test technology. FLS function was evaluated using a glycolysis inhibitor, 2-deoxy-d-glucose (2-DG). After stimulation of the FLS, a migration scratch assay, MTT assay, and enzyme-linked immunosorbent assay were performed to measure the effect of 2-DG on FLS migration, viability of the FLS, and cytokine secretion, respectively. IRDye 800CW 2-DG was used to assess glucose uptake in the arthritic joints and stromal cells of mice after K/BxN mouse serum transfer. The mice were injected daily, intraperitoneally, with 3-bromopyruvate (BrPa; 5 mg/kg) to assess the effect of inhibition of glycolysis in vivo.ResultsCompared to human OA FLS, the balance between glycolysis and oxidative phosphorylation was shifted toward glycolysis in RA FLS. Glucose transporter 1 (GLUT1) messenger RNA (mRNA) expression correlated with baseline functions of the RA FLS. Glucose deprivation or incubation of the FLS with glycolytic inhibitors impaired cytokine secretion and decreased the rate of proliferation and migration of the cells. In a mouse model of inflammatory arthritis, GLUT1 mRNA expression in the synovial lining cells was observed, and increased levels of glucose uptake and glycolytic gene expression were detected in the stromal compartment of the arthritic mouse joints. Inhibition of glycolysis by BrPa, administered in vivo, significantly decreased the severity of arthritis in this mouse model.ConclusionTargeting metabolic pathways is a novel approach to understanding the mechanisms of disease. Inhibition of glycolysis may directly modulate synoviocyte-mediated inflammatory functions and could be an effective treatment strategy for arthritis.

Dataset Information

Downregulation of lncRNA ITSN1-2 correlates with decreased disease risk and activity of rheumatoid arthritis (RA), and reduces RA fibroblast-like synoviocytes proliferation and inflammation via inhibiting NOD2/RIP2 signaling pathway.

Publications

Downregulation of lncRNA ITSN1-2 correlates with decreased disease risk and activity of rheumatoid arthritis (RA), and reduces RA fibroblast-like synoviocytes proliferation and inflammation via inhibiting NOD2/RIP2 signaling pathway.

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