Cacciapaglia Fabio, Perniola Simone, Stano Stefano, Venerito Vincenzo, Natuzzi Dorotea, Bizzoca Rita, Iannone Florenzo
Rheumatology Unit, Department of Precision and Regenerative Medicine and Ionian Area (DiMePre-J), University of Bari, Bari, Italy; Department of Medicine and Surgery, LUM University "Giuseppe De Gennaro" Casamassima & Rheumatology Service "Miulli" General Hospital Acquaviva delle Fonti, Bari, Italy.
Rheumatology Unit, Department of Precision and Regenerative Medicine and Ionian Area (DiMePre-J), University of Bari, Bari, Italy.
Exp Mol Pathol. 2025 Mar;141:104951. doi: 10.1016/j.yexmp.2024.104951. Epub 2024 Dec 25.
Interleukin-6 (IL-6) is a relevant cytokine in rheumatoid arthritis (RA) pathogenesis, potentially activating Janus kinases (JAK)-1, -2, and tyrosine kinase 2 (TYK2), and thus, three signal transducer and activator of transcription (STAT)-1, -3 or - 5 pathways. This pilot study aims to explore differences in phosphorylated (p)STAT3 levels among patients with RA, those not classified as RA (nRA), and healthy donors (HD), providing some clues on the relative contribution of each JAK protein to the downstream of the IL-6-induced STAT3 pathway. Clinical data and blood samples from 80 subjects (41 RA, 14 nRA, and 25 HD) were collected. The activity of the JAK-STAT3 pathway was assessed by Western Blot and Real Time-PCR analysis for the quantification of STAT3 in peripheral blood mononuclear cells (PBMC). Furthermore, the impact of JAK-1, -2, and TYK2 inhibitors on pSTAT3 was assessed in vitro by FACS, with and without IL-6 stimulation in RA patients naïve to treatment with DMARD and steroids. The pSTAT3 (%) was significantly higher in PBMC from RA compared to nRA patients and HD. Furthermore, pSTAT3 (%) was significantly associated with inflammation and disease activity (ESR, CRP, and DAS28). The JAK-1 inhibitor was more effective in reducing pSTAT3 expression in CD14 cells of RA patients, while the JAK-2 selective compound was more effective in CD4 cells of RA patients. On the contrary, the TYK2 selective agent showed no significant effects. This study highlights the importance of the JAK/STAT3 pathway in RA. Some differences among various JAK proteins have been pointed out, with JAK1 and JAK2 standing as the most relevant mediators of the STAT3 pathway in this in-vitro model after IL-6R activation.
白细胞介素-6（IL-6）是类风湿关节炎（RA）发病机制中的一种相关细胞因子，可能激活Janus激酶（JAK）-1、-2和酪氨酸激酶2（TYK2），进而激活三种信号转导和转录激活因子（STAT）-1、-3或-5信号通路。这项初步研究旨在探讨RA患者、未分类为RA的患者（nRA）和健康供体（HD）之间磷酸化（p）STAT3水平的差异，为每种JAK蛋白对IL-6诱导的STAT3信号通路下游的相对贡献提供一些线索。收集了80名受试者（41名RA患者、14名nRA患者和25名HD）的临床数据和血液样本。通过蛋白质免疫印迹法（Western Blot）和实时定量聚合酶链反应（Real Time-PCR）分析评估JAK-STAT3信号通路的活性，以定量外周血单个核细胞（PBMC）中的STAT3。此外，在未接受过改善病情抗风湿药（DMARD）和类固醇治疗的RA患者中，通过流式细胞术（FACS）在有或无IL-6刺激的情况下，体外评估JAK-1、-2和TYK2抑制剂对pSTAT3的影响。与nRA患者和HD相比，RA患者PBMC中的pSTAT3（%）显著更高。此外，pSTAT3（%）与炎症和疾病活动（红细胞沉降率、C反应蛋白和疾病活动评分28 [DAS28]）显著相关。JAK-1抑制剂在降低RA患者CD14细胞中pSTAT3表达方面更有效，而JAK-2选择性化合物在RA患者CD4细胞中更有效。相反，TYK2选择性药物未显示出显著效果。这项研究突出了JAK/STAT3信号通路在RA中的重要性。指出了各种JAK蛋白之间的一些差异，在该体外模型中，IL-6受体激活后，JAK1和JAK2是STAT3信号通路最相关的介质。
