Center for Integrative Rheumatoid Transcriptomics and Dynamics, The Catholic University of Korea, Seoul 06591, Korea.
Division of Urology, Department of Surgery and Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, CA 90048.
J Immunol. 2018 Jul 15;201(2):359-370. doi: 10.4049/jimmunol.1701097. Epub 2018 May 23.
Fibroblast-like synoviocytes (FLSs) play a key role in the progression of rheumatoid arthritis (RA) as a primary component of invasive hypertrophied pannus. FLSs of RA patients (RA-FLSs) exhibit cancer-like features, including promigratory and proinvasive activities that largely contribute to joint cartilage and bone destruction. In this study, we hypothesized that the NF of activated T cell 5 (NFAT5), a transcription factor involving tumor invasiveness, would control the migration and invasion of RA-FLSs. Analyses of transcriptomes demonstrated the significant involvement of NFAT5 in locomotion of RA-FLSs and that tissue factor (TF; also known as coagulation factor III) and CCL2 were the major downstream target genes of NFAT5 involving FLS migration and invasion. In cultured RA-FLSs, IL-1β and TGF-β increased TF and CCL2 expression by upregulating NFAT5 expression via p38 MAPK. Functional assays demonstrated that NFAT5- or TF-deficient RA-FLSs displayed decreased lamellipodia formation, cell migration, and invasion under IL-1β- or TGF-β-stimulated conditions. Conversely, factor VIIa, a specific activator of TF, increased migration of RA-FLSs, which was blocked by NFAT5 knockdown. Recombinant CCL2 partially restored the decrease in migration and invasion of NFAT5-deficient RA-FLSs stimulated with IL-1β. NFAT5-knockout mouse FLSs also showed decreased expressions of TF and CCL2 and reduced cell migration. Moreover, KRN2, a specific inhibitor of NFAT5, suppressed migration of FLSs stimulated with TGF-β. Conclusively, to our knowledge, this is the first study to provide evidence of a functional link between osmoprotective NFAT5 and TF in the migration and invasion of RA-FLSs and supports a role for NFAT5 blockade in the treatment of RA.
成纤维样滑膜细胞（FLS）作为侵袭性肥大血管翳的主要组成部分，在类风湿关节炎（RA）的进展中起着关键作用。RA 患者的 FLS（RA-FLS）表现出类似癌症的特征，包括促迁移和促侵袭活性，这在很大程度上导致关节软骨和骨破坏。在这项研究中，我们假设活化 T 细胞核因子 5（NFAT5），一种涉及肿瘤侵袭性的转录因子，将控制 RA-FLS 的迁移和侵袭。转录组分析表明 NFAT5 显着参与 RA-FLS 的运动，组织因子（TF；也称为凝血因子 III）和 CCL2 是涉及 FLS 迁移和侵袭的 NFAT5 的主要下游靶基因。在培养的 RA-FLS 中，IL-1β 和 TGF-β 通过上调 p38 MAPK 来增加 NFAT5 表达，从而增加 TF 和 CCL2 的表达。功能测定表明，在 IL-1β 或 TGF-β 刺激下，NFAT5 或 TF 缺陷型 RA-FLS 的片状伪足形成、细胞迁移和侵袭减少。相反，TF 的特异性激活物因子 VIIa 增加了 RA-FLS 的迁移，而 NFAT5 敲低则阻断了这种迁移。重组 CCL2 部分恢复了 NFAT5 缺陷型 RA-FLS 在 IL-1β 刺激下迁移和侵袭的减少。NFAT5 敲除小鼠 FLS 也表现出 TF 和 CCL2 的表达减少和细胞迁移减少。此外，NFAT5 的特异性抑制剂 KRN2 抑制了 TGF-β 刺激的 FLS 迁移。总之，据我们所知，这是第一项提供证据表明在 RA-FLS 的迁移和侵袭中，具有渗透保护作用的 NFAT5 和 TF 之间存在功能联系，并支持 NFAT5 阻断在 RA 治疗中的作用的研究。
