Vomero M, Caliste M, Barbati C, Speziali M, Celia A I, Ucci F, Ciancarella C, Putro E, Colasanti T, Buoncuore G, Corsiero E, Bombardieri M, Spinelli F R, Ceccarelli F, Conti F, Alessandri C
Arthritis Center, Dipartimento di Scienze Cliniche Internistiche, Anestesiologiche e Cardiovascolari, Sapienza University of Rome, Rome, Italy.
Rheumatology, Immunology and Clinical Medicine Unit, Università Campus Bio-Medico di Roma, Rome, Italy.
Front Pharmacol. 2022 Mar 3;13:852802. doi: 10.3389/fphar.2022.852802. eCollection 2022.
The pathway of Janus tyrosine kinases (JAKs) has a central role in the pathogenesis of Rheumatoid Arthritis (RA) by regulating multiple immune functions and cytokine production. The JAK inhibitor tofacitinib is effective in RA patients not responding to methotrexate or TNF-inhibitors. Since hyperactive autophagy has been associated with impaired apoptosis of RA fibroblast-like synoviocytes (FLS), we aimed to investigate the role of tofacitinib in modulating autophagy and apoptosis in these cells. FLS isolated from RA biopsies were cultured with tofacitinib in presence of autophagy inducer rapamycin and in serum deprivation condition. Levels of autophagy, apoptosis, and citrullinated proteins were analyzed by western blot, flow cytometry, immunocytofluorescence, and Real-Time PCR. Rapamycin induced an increase in RA-FLS autophagy while the levels of autophagy marker LC3-II were reduced after treatment with tofacitinib. The analysis of autophagic flux by specific fluorescence dye confirmed the reduction of autophagy in RA FLS. The treatment with tofacitinib did not influence apoptosis of RA FLS. Modulation of the autophagic process by tofacitinib did not significantly change citrullination. The results of this study demonstrate that tofacitinib is able to modulate autophagy of FLS contributing to its effectiveness in RA patients.
Janus酪氨酸激酶（JAK）通路通过调节多种免疫功能和细胞因子产生，在类风湿关节炎（RA）的发病机制中起核心作用。JAK抑制剂托法替布对甲氨蝶呤或肿瘤坏死因子（TNF）抑制剂治疗无效的RA患者有效。由于自噬过度活跃与RA成纤维细胞样滑膜细胞（FLS）凋亡受损有关，我们旨在研究托法替布在调节这些细胞自噬和凋亡中的作用。从RA活检组织中分离出的FLS在自噬诱导剂雷帕霉素存在及血清饥饿条件下，用托法替布进行培养。通过蛋白质免疫印迹法、流式细胞术、免疫细胞荧光法和实时聚合酶链反应分析自噬、凋亡和瓜氨酸化蛋白水平。雷帕霉素诱导RA - FLS自噬增加，而用托法替布治疗后自噬标志物LC3 - II水平降低。通过特异性荧光染料分析自噬通量证实RA FLS中自噬减少。托法替布治疗不影响RA FLS的凋亡。托法替布对自噬过程的调节未显著改变瓜氨酸化。本研究结果表明，托法替布能够调节FLS的自噬，这有助于其对RA患者的疗效。
