Sportology Center, Juntendo University, Tokyo, Japan.
Department of Pathophysiology for Locomotive and Neoplastic Diseases, Juntendo University Graduate School of Medicine, Tokyo, Japan.
Lab Invest. 2022 Jan;102(1):102-111. doi: 10.1038/s41374-021-00685-4. Epub 2021 Oct 30.
The ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) family includes nine members with aggrecan-degrading activity, i.e., ADAMTS1, 4, 5, 8, 9, 15, 16, 18, and 20. However, their systematic expression profile in knee osteoarthritis (OA) synovium and effects of cytokines and growth factors on the expression in OA synovial fibroblasts remain elusive. In this study, expression of all nine aggrecanolytic ADAMTS species was assessed by quantitative real-time PCR in OA and control normal synovial tissues. OA synovial fibroblasts were treated with interleukin-1α (IL-1α), IL-1β, tumor necrosis factor-α (TNF-α), transforming growth factor-β (TGF-β), vascular endothelial growth factor, and heparin-binding epidermal growth factor, and analyzed for the expression of the ADAMTS species. The signaling pathways and inhibition of ADAMTS4 expression by high-molecular-weight hyaluronan, adalimumab, tocilizumab, and signaling molecule inhibitors were studied. ADAMTS1, 4, 5, 9, and 16 were expressed in OA synovium, but only ADAMTS4 expression was significantly higher in OA as compared to normal synovium. IL-1α, TNF-α, and TGF-β markedly increased ADAMTS4 expression, while their effects were minimal for the other ADAMTS species. ADAMTS4 was synergistically upregulated by treatment with IL-1α and TNF-α, IL-1α and TGF-β, or IL-1α, TNF-α and TGF-β. The signaling molecules' inhibitors demonstrated that IL-1α-induced ADAMTS4 expression is predominantly through TGF-β-associated kinase 1 (TAK1), and the TNF-α-stimulated expression is via TAK1 and nuclear factor-κB (NF-κB). The TGF-β-promoted expression was through the activin receptor-like kinase 5 (ALK5)/Smad2/3, TAK1, and non-TAK1 pathways. Adalimumab blocked TNF-α-stimulated expression. ADAMTS4 expression co-stimulated with IL-1α, TNF-α and TGF-β was abolished by treatment with adalimumab, TAK1 inhibitor, and ALK5/Smad2/3 inhibitor. These data demonstrate marked and synergistic upregulation of ADAMTS4 by IL-1α, TNF-α and TGF-β in OA synovial fibroblasts, and suggest that concurrent therapy with an anti-TNF-α drug and inhibitor(s) may be useful for prevention against aggrecan degradation in OA.
ADAMTS（解整合素和金属蛋白酶与凝血酶反应蛋白域）家族包括 9 种具有聚集蛋白降解活性的成员，即 ADAMTS1、4、5、8、9、15、16、18 和 20。然而，它们在膝骨关节炎（OA）滑膜中的系统表达谱以及细胞因子和生长因子对 OA 滑膜成纤维细胞表达的影响仍不清楚。在这项研究中，通过定量实时 PCR 评估了 OA 和对照正常滑膜组织中所有 9 种聚集蛋白降解 ADAMTS 种的表达。用白细胞介素 1α（IL-1α）、白细胞介素 1β（IL-1β）、肿瘤坏死因子-α（TNF-α）、转化生长因子-β（TGF-β）、血管内皮生长因子和肝素结合表皮生长因子处理 OA 滑膜成纤维细胞，并分析 ADAMTS 种的表达。研究了 ADAMTS4 表达的信号通路和高相对分子质量透明质酸、阿达木单抗、托珠单抗和信号分子抑制剂的抑制作用。ADAMTS1、4、5、9 和 16 在 OA 滑膜中表达，但只有 ADAMTS4 的表达在 OA 中明显高于正常滑膜。IL-1α、TNF-α 和 TGF-β 显著增加 ADAMTS4 的表达，而对其他 ADAMTS 种的影响最小。用 IL-1α 和 TNF-α、IL-1α 和 TGF-β 或 IL-1α、TNF-α 和 TGF-β 联合处理可协同上调 ADAMTS4 的表达。信号分子抑制剂表明，IL-1α 诱导的 ADAMTS4 表达主要通过 TGF-β 相关激酶 1（TAK1），而 TNF-α 刺激的表达则通过 TAK1 和核因子-κB（NF-κB）。TGF-β 促进的表达是通过激活素受体样激酶 5（ALK5）/Smad2/3、TAK1 和非 TAK1 途径。阿达木单抗阻断 TNF-α 刺激的表达。用阿达木单抗、TAK1 抑制剂和 ALK5/Smad2/3 抑制剂处理可消除 ADAMTS4 与 IL-1α、TNF-α 和 TGF-β 共刺激的表达。这些数据表明，IL-1α、TNF-α 和 TGF-β 在 OA 滑膜成纤维细胞中显著协同上调 ADAMTS4 的表达，并提示同时使用抗 TNF-α 药物和（或）抑制剂可能有助于预防 OA 中聚集蛋白的降解。
