Targeting non-canonical nuclear factor-&kgr;B signalling attenuates neovascularization in a novel 3D model of rheumatoid arthritis synovial angiogenesis
Targeting non-canonical nuclear factor-&kgr;B signalling attenuates neovascularization in a novel 3D model of rheumatoid arthritis synovial angiogenesis
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针对非典型核因子-
作者：
C. Maracle;P. Kucharzewska;B. Helder;C. van der Horst;P. Correa de Sampaio;A. Noort;K. V. van Zoest;A. Griffioen;H. Olsson;S. Tas
目标。血管生成是RA疾病进展的关键。淋巴毒素bgr;受体（LT&bgr；R）诱导的非规范核因子-&kgr的激活；B （NF-&kgr；B）途径通过NF-&kgr；b诱导激酶（NIK）参与了这一过程。因此，抑制这一途径可能具有治疗RA的潜力。我们描述了一个新的三维（3D）滑膜血管生成模型，包括内皮细胞（ECs）、RA成纤维细胞样滑膜细胞（RAFLSs）和RA滑膜液（RASF），以进一步研究NF-&kgr的作用；B在这个过程中。方法。由RAFLSs和ECs组成的球体被RASF刺激，LT&bgr；R配体LT&bgr；和LIGHT，或生长因子bFGF和VEGF，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，anginex、靶向nik的siRNA （siNIK）、ltp的作用；研究了R-Ig融合蛋白（baminerept）和一种新型药理NIK抑制剂。结果。RASF显著促进了血管生成抑制剂anginex对芽形成的抑制作用（P < 0.05）。LT&bgr;光诱导显著发芽（P < 0.05）， bFGF/VEGF诱导显著发芽（P < 0.01）。对ECs进行siNIK预处理导致LT&bgr降低；r诱导血管形成（P < 0.05）。LT&bgr;R-Ig不仅阻断了LT&bgr；-或光诱导发芽，但rasf诱导发芽也有差异（P < 0.05）。NIK抑制剂阻断LT&bgr诱导的血管生成；、LIGHT、生长因子（P < 0.05）和RASF （P < 0.01）。结论。我们提出了一个新的三维滑膜血管生成模型，包括RAFLSs， ECs和RASF，模拟体内情况。利用该系统，我们证明了非正则NF-&kgr；B信号传导促进新生血管形成，并表明该模型可用于解剖特定细胞类型中信号传导途径对血管生成反应的相对贡献，并用于测试血管生成的药理学抑制剂。
Objective. Angiogenesis is crucial in RA disease progression. Lymphotoxin &bgr; receptor (LT&bgr;R)-induced activation of the non-canonical nuclear factor-&kgr;B (NF-&kgr;B) pathway via NF-&kgr;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-&kgr;B in this process. Methods. Spheroids consisting of RAFLSs and ECs were stimulated with RASF, the LT&bgr;R ligands LT&bgr; 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&bgr;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&bgr; 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&bgr;R-induced vessel formation (P < 0.05). LT&bgr;R–Ig not only blocked LT&bgr;- or LIGHT-induced sprouting, but also RASF-induced sprouting (P < 0.05). The NIK inhibitor blocked angiogenesis induced by LT&bgr;, 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-&kgr;B 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.
DOI：
10.1073/pnas.92.10.4342
发表时间：
1995-05-09
影响因子：
11.1
作者：
KARMANN, K;HUGHES, CCW;POBER, JS
通讯作者：
POBER, JS