类风湿性关节炎（RA）是一种以免疫稳态失衡为特征的自身免疫性炎症疾病。 p53 突变体通常被描述为癌细胞的守护者，赋予癌细胞耐药性和免疫逃避能力。重要的是，在 RA 患者中也发现了 p53 突变，这促使人们研究其在 RA 发病机制中的作用。缓解疾病抗风湿药物 (DMARD) 对 p53 野生型 (WT)/突变体转染的 RA 的细胞毒性通过MTT测定评估成纤维细胞样滑膜细胞（RAFLS）。采用腺相关病毒（AAV）建立p53 WT/R211*佐剂诱导性关节炎（AIA）大鼠模型。通过显微 CT 分析等各种参数评估大鼠的关节炎状况。分离膝关节样本进行总RNA测序分析。通过qPCR、ELISA检测和免疫荧光检测细胞因子和免疫相关基因的表达。通过体外和体内免疫沉淀和蛋白质印迹确定其机制途径。在p53突变体中，p53R213*在RAFLS中表现出显着的DMARD抗性。然而，AAV 诱导的 p53R211* 过表达可改善不具有甲氨蝶呤 (MTX) 耐药性的 AIA 大鼠的炎症性关节炎，我们的结果发现 p53R211* 通过抑制大鼠中 T 细胞活化和 T 辅助 17 细胞 (Th17) 浸润发挥免疫调节作用联合，最终下调促炎细胞因子的表达。总 RNA 测序分析确定了 p53R211* 与免疫相关通路的相关性。进一步的机制研究表明，p53R213*/R211*（而不是野生型 p53）与 TANK 结合激酶 1 (TBK1) 相互作用，并抑制先天免疫 TBK1-干扰素调节因子 3 (IRF3)-干扰素基因刺激剂 (STING) 级联。这项研究揭示了 p53R213* 突变体在 RA 发病机制中的作用，并将 TBK1 确定为潜在的抗炎靶点。© 2023。作者。

Rheumatoid arthritis (RA) is an autoimmune inflammation disease characterized by imbalance of immune homeostasis. p53 mutants are commonly described as the guardian of cancer cells by conferring them drug-resistance and immune evasion. Importantly, p53 mutations have also been identified in RA patients, and this prompts the investigation of its role in RA pathogenesis.The cytotoxicity of disease-modifying anti-rheumatic drugs (DMARDs) against p53 wild-type (WT)/mutant-transfected RA fibroblast-like synoviocytes (RAFLSs) was evaluated by MTT assay. Adeno-associated virus (AAV) was employed to establish p53 WT/R211* adjuvant-induced arthritis (AIA) rat model. The arthritic condition of rats was assessed by various parameters such as micro-CT analysis. Knee joint samples were isolated for total RNA sequencing analysis. The expressions of cytokines and immune-related genes were examined by qPCR, ELISA assay and immunofluorescence. The mechanistic pathway was determined by immunoprecipitation and Western blotting in vitro and in vivo.Among p53 mutants, p53R213* exhibited remarkable DMARD-resistance in RAFLSs. However, AAV-induced p53R211* overexpression ameliorated inflammatory arthritis in AIA rats without Methotrexate (MTX)-resistance, and our results discovered the immunomodulatory effect of p53R211* via suppression of T-cell activation and T helper 17 cell (Th17) infiltration in rat joint, and finally downregulated expressions of pro-inflammatory cytokines. Total RNA sequencing analysis identified the correlation of p53R211* with immune-related pathways. Further mechanistic studies revealed that p53R213*/R211* instead of wild-type p53 interacted with TANK-binding kinase 1 (TBK1) and suppressed the innate immune TBK1-Interferon regulatory factor 3 (IRF3)-Stimulator of interferon genes (STING) cascade.This study unravels the role of p53R213* mutant in RA pathogenesis, and identifies TBK1 as a potential anti-inflammatory target.© 2023. The Author(s).