Targeting non-canonical nuclear factor-κB signalling attenuates neovascularization in a novel 3D model of rheumatoid arthritis synovial angiogenesis
Targeting non-canonical nuclear factor-κB signalling attenuates neovascularization in a novel 3D model of rheumatoid arthritis synovial angiogenesis
复制标题
在一种新的类风湿性关节炎滑膜血管生成3D模型中，靶向非典型核因子-κB信号可减弱新生血管
DOI：
10.1093/rheumatology/kew393
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发表时间：
2017-02-01
期刊：
影响因子：
5.5
通讯作者：
Tas, Sander W.
中科院分区：
文献类型：
--
作者：
Maracle, Chrissta X.;Kucharzewska, Paulina;Tas, Sander W.
Objective.血管生成在RA疾病进展中至关重要。在此过程中涉及到非典型核因子-κ B（NF-κ B）途径通过NF-κ B-诱导激酶（NIK）的光敏素β受体（LT β R）诱导的活化。因此，抑制该途径可能在RA中具有治疗潜力。我们描述了一种新的三维（3D）滑膜血管生成模型，包括内皮细胞（EC）、RA成纤维细胞样滑膜细胞（RAFLS）和RA滑液（RASF），以进一步研究NF-κ B在此过程中的作用。用RASF、LT β R配体LT β和LIGHT或生长因子bFGF和VEGF刺激由RAFLS和EC组成的球体，然后使用共聚焦显微镜和数字图像分析定量EC发芽。接下来，研究了anginex、NIK靶向siRNA（siNIK）、LT beta R-Ig融合蛋白（baminercept）和新型药理学NIK抑制剂的作用。RASF能显著促进芽的形成，而血管生成抑制剂anginex能显著抑制芽的形成（P < 0.05）。LT β和LIGHT诱导了显著的芽生（P < 0.05），bFGF/ VEGF也诱导了显著的芽生（P < 0.01）。siNIK预处理EC可显著降低LT β R诱导的血管形成（P < 0.05）。LT β R -IG不仅能阻断LT β或LIGHT诱导的芽生，而且能阻断RASF诱导的芽生（P < 0.05）。NIK抑制剂可阻断LT β、LIGHT、生长因子（P< 0.05）和RASF诱导的血管生成（P < 0.01）。我们提出了一种新的三维模型，滑膜血管生成RAFLSs，ECs和RASF，模仿体内的情况。使用这个系统，我们证明了非经典的NF-κ B信号促进新血管形成，并表明，该模型是有用的解剖在特定的细胞类型的血管生成反应的信号通路的相对贡献和测试血管生成的药理学抑制剂。
Objective. Angiogenesis is crucial in RA disease progression. Lymphotoxin beta receptor (LT beta R)-induced activation of the non-canonical nuclear factor-kappa B (NF-kappa B) pathway via NF-kappa B-inducing kinase (NIK) has been implicated in this process. Consequently, inhibition of this pathway may hold therapeutic potential in RA. We describe a novel three-dimensional (3D) model of synovial angiogenesis incorporating endothelial cells (ECs), RA fibroblast-like synoviocytes (RAFLSs) and RA synovial fluid (RASF) to further investigate the contributions of NF-kappa B in this process.Methods. Spheroids consisting of RAFLSs and ECs were stimulated with RASF, the LT beta R ligands LT beta and LIGHT, or growth factor bFGF and VEGF, followed by quantification of EC sprouting using confocal microscopy and digital image analysis. Next, the effects of anginex, NIK-targeting siRNA (siNIK), LT beta R-Ig fusion protein (baminercept) and a novel pharmacological NIK inhibitor were investigated.Results. RASF significantly promoted sprout formation, which was blocked by the established angiogenesis inhibitor anginex (P < 0.05). LT beta and LIGHT induced significant sprouting (P < 0.05), as did bFGF/ VEGF (P < 0.01). siNIK pre-treatment of ECs led to reductions in LT beta R-induced vessel formation (P < 0.05). LT beta R -Ig not only blocked LT beta-or LIGHT-induced sprouting, but also RASF-induced sprouting (P < 0.05). The NIK inhibitor blocked angiogenesis induced by LT beta, LIGHT, growth factors (P< 0.05) and RASF (P < 0.01).Conclusion. We present a novel 3D model of synovial angiogenesis incorporating RAFLSs, ECs and RASF that mimics the in vivo situation. Using this system, we demonstrate that non-canonical NF-kB signalling promotes neovascularization and show that this model is useful for dissecting relative contributions of signalling pathways in specific cell types to angiogenic responses and for testing pharmacological inhibitors of angiogenesis.