Department of Orthopedic Surgery, Kobe University Graduate School of Medicine, 7-5-1, Kusunoki-Cho, Chuo-Ku, Kobe, Hyogo, 650-0017, Japan.
Department of Orthopedic Surgery, Matsubara Mayflower Hospital, 944-25, Fujita, Katō, Hyogo, Japan.
Clin Rheumatol. 2024 Nov;43(11):3525-3536. doi: 10.1007/s10067-024-07142-9. Epub 2024 Sep 20.
INTRODUCTION/OBJECTIVES: JAK/STAT signaling inhibition exerts therapeutic effects on angiogenesis in rheumatoid arthritis (RA). However, whether the inhibitory effect differs among JAK inhibitors because of differing selectivity is unknown. Therefore, we compared the inhibitory effects of tofacitinib, baricitinib, peficitinib, upadacitinib, and filgotinib on angiogenesis.
RA-derived fibroblast-like synoviocytes (RA-FLS) were seeded on type I collagen gel, and human umbilical vein endothelial cells (HUVECs) were directly added. The control and aforementioned JAK inhibitors were added to the medium, followed by stimulation with interleukin (IL)-6 and soluble IL-6 receptor (sIL-6R). Each JAK inhibitor's concentration was determined based on estimated blood concentrations. The vascular endothelial growth factor (VEGF) concentration was evaluated with an enzyme-linked immunosorbent assay using the medium from the first exchange. A migration assay was performed, and HUVEC migration was evaluated using CD31 fluorescence immunostaining.
Hematoxylin-eosin staining showed that compared with the non-JAKi treatment group, the JAKi treatment group markedly degenerated in the sub-lining and deep lining, with decreased lymphocyte infiltration and neovascularization [Rooney's score subscale, non-JAKi vs JAKi (median, 6.5 vs 2.5, p = 0.005)]. In vitro, IL-6 and sIL-6R administration increased VEGF production from RA-FLS and promoted neovascularization in HUVECs, and JAK-inhibitor administration, which decreased VEGF production from RA-FLS and suppressed HUVEC migration, inhibited neovascularization in RA-FLS and HUVEC co-cultures.
The JAK inhibitors suppressed IL-6-induced angiogenesis via decreased VEGF production and HUVEC migration in RA-FLS and HUVEC co-cultures. No significant differences were observed among the JAK inhibitors, whose anti-angiogenic effect may be an important mechanism for RA treatment. Key Points • JAK inhibitors inhibit angiogenesis in RA by reducing VEGF production from RA-derived fibroblast-like synoviocytes. • Our study provides new insights into RA treatment by elucidating the anti-angiogenic effect of JAK inhibitors.
简介/目的：JAK/STAT 信号抑制对类风湿关节炎（RA）中的血管生成具有治疗作用。然而，由于选择性不同，不同 JAK 抑制剂的抑制效果是否不同尚不清楚。因此，我们比较了托法替尼、巴瑞替尼、培非替尼、乌帕替尼和菲戈替尼对血管生成的抑制作用。
将 RA 衍生的成纤维样滑膜细胞（RA-FLS）接种在 I 型胶原凝胶上，并直接添加人脐静脉内皮细胞（HUVEC）。将对照和上述 JAK 抑制剂添加到培养基中，然后用白细胞介素（IL）-6 和可溶性 IL-6 受体（sIL-6R）刺激。根据估计的血液浓度确定每种 JAK 抑制剂的浓度。用酶联免疫吸附试验（ELISA）评估培养基中的血管内皮生长因子（VEGF）浓度，第一次交换时使用。进行迁移试验，并用 CD31 荧光免疫染色评估 HUVEC 迁移。
苏木精-伊红染色显示，与非 JAKi 治疗组相比，JAKi 治疗组在衬里下层和深层衬里明显退化，淋巴细胞浸润和新生血管减少[罗尼评分亚量表，非 JAKi 与 JAKi（中位数，6.5 与 2.5，p=0.005)]。在体外，IL-6 和 sIL-6R 给药增加了 RA-FLS 中 VEGF 的产生，并促进了 HUVEC 中的新生血管形成，而 JAK 抑制剂给药减少了 RA-FLS 中 VEGF 的产生并抑制了 HUVEC 的迁移，抑制了 RA-FLS 和 HUVEC 共培养物中的新生血管形成。
JAK 抑制剂通过减少 RA-FLS 和 HUVEC 共培养物中 VEGF 的产生和 HUVEC 迁移来抑制 IL-6 诱导的血管生成。JAK 抑制剂之间没有观察到显著差异，其抗血管生成作用可能是 RA 治疗的一个重要机制。关键点：JAK 抑制剂通过减少 RA 衍生的成纤维样滑膜细胞中 VEGF 的产生来抑制 RA 中的血管生成。我们的研究通过阐明 JAK 抑制剂的抗血管生成作用，为 RA 治疗提供了新的见解。
