Department of Infection Immunology, Tianjin Hospital, No 406 South Jiefang Road, Hexi District, Tianjin, 300221, China.
J Bioenerg Biomembr. 2024 Oct;56(5):563-571. doi: 10.1007/s10863-024-10035-w. Epub 2024 Aug 27.
Rheumatoid arthritis (RA) is a chronic condition characterized by inflammation and an abnormal immune response. N6-methyladenosine (m6A) methylation has altered nucleotide-binding oligomerization domain, leucine-rich repeat, and pyrin domain-containing (NLRP) 3. This change is implicated in the regulation of cell pyroptosis and inflammation. WTAP has a crucial role in regulating NLRP3 m6A. In this work, we used a rat model of collagen-induced arthritis (CIA) to investigate the involvement of WTAP in the evolution of inflammation in RA. The purpose of silencing or overexpressing WTAP in RA-fibroblast-like synoviocytes (RA-FLSs) treated with TNF-α was to identify its impact on pyroptosis, NLRP3 inflammasome-related proteins, the secretion of pro-inflammatory cytokines and migration. Bioinformatics techniques were used to pinpoint the exact target controlled by WTAP. To assess WTAP and NLRP3's role in RA-FLSs, we used methylated RNA immunoprecipitation, LDH test, flow cytometry, RT-qPCR, Western blotting, and Transwell. Our results show that WTAP expression is upregulated in both RA rats and cell models. Cell pyroptosis, NLRP3-related pro-inflammatory cytokines, and migration were reduced in TNF-α-treated RA-FLSs when WTAP was knocked down, whereas overexpression of WTAP displayed the opposite effect in RA-FLSs. WTAP mediated m6A modification in the NLRP3 mRNA and enhanced its mRNA stability. These results suggested that WTAP promoted FLSs pyroptosis and related inflammatory response via NLRP3 and identified WTAP as a potential target for treating RA.
类风湿关节炎(RA)是一种以炎症和异常免疫反应为特征的慢性疾病。N6-甲基腺苷(m6A)甲基化改变了核苷酸结合寡聚化结构域、富含亮氨酸重复和吡咯啉域包含(NLRP)3。这种变化与细胞焦亡和炎症的调节有关。WTAP 在调节 NLRP3 m6A 中起着至关重要的作用。在这项工作中,我们使用胶原诱导性关节炎(CIA)大鼠模型来研究 WTAP 在 RA 炎症演变中的作用。沉默或过表达 TNF-α 处理的 RA 成纤维样滑膜细胞(RA-FLS)中的 WTAP,以确定其对细胞焦亡、NLRP3 炎性体相关蛋白、促炎细胞因子分泌和迁移的影响。生物信息学技术用于确定 WTAP 控制的确切靶点。为了评估 WTAP 和 NLRP3 在 RA-FLS 中的作用,我们使用了甲基化 RNA 免疫沉淀、LDH 试验、流式细胞术、RT-qPCR、Western blot 和 Transwell。我们的结果表明,WTAP 在 RA 大鼠和细胞模型中均上调表达。当 WTAP 被敲低时,TNF-α 处理的 RA-FLS 中的细胞焦亡、NLRP3 相关促炎细胞因子和迁移减少,而 RA-FLS 中 WTAP 的过表达则表现出相反的效果。WTAP 介导 NLRP3 mRNA 的 m6A 修饰并增强其 mRNA 稳定性。这些结果表明,WTAP 通过 NLRP3 促进 FLSs 焦亡和相关炎症反应,并将 WTAP 确定为治疗 RA 的潜在靶点。