Actin remodeling by Nck regulates endothelial lumen formation.
Actin remodeling by Nck regulates endothelial lumen formation.
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DOI：
10.1091/mbc.e15-06-0338
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发表时间：
2015-09-01
影响因子：
3.3
通讯作者：
Rivera GM
中科院分区：
文献类型：
--
作者：
Chaki SP;Barhoumi R;Rivera GM
NCK依赖的肌动蛋白重塑通过促进细胞伸长和VE-钙粘蛋白细胞间连接的适当组织来实现内皮形态发生。Nck决定Cdc 42/aPKC激活的时空模式，以调节内皮细胞的顶部-底部极性和管腔形成。多种血管生成信号调节磷酸酪氨酸信号以促进血管发生和血管生成。尽管其功能和临床的重要性，血管细胞如何整合磷酸酪氨酸依赖性信号，以引起内皮形态发生所需的细胞骨架的变化仍然知之甚少。Nck衔接子家族将磷酸酪氨酸信号与肌动蛋白动力学偶联，因此能够很好地协调血管形成和重塑所需的细胞过程。在VEGF刺激存在下，在三维胶原基质中培养内皮细胞与分子遗传学、光学成像和生物化学相结合，以显示NCK依赖性肌动蛋白重塑促进内皮细胞伸长和VE-钙粘蛋白细胞间连接的适当组织。主要形态发生缺陷所造成的废除NCK信号包括损失的内皮细胞的顶部-基底极性和受损的管腔化。使用Förster共振能量转移生物传感器的延时成像，磷酸化特异性抗体的免疫染色和GST下拉分析表明，Nck决定了内皮形态发生过程中Cdc 42/aPKC激活的时空模式。我们的研究结果表明，NCK作为一个重要的枢纽整合血管生成线索与细胞骨架的变化，使内皮细胞的顶端-基底极化和管腔形成。这些发现指出Nck是有效抗血管生成治疗的紧急靶点。
Nck-dependent actin remodeling enables endothelial morphogenesis by promoting cell elongation and proper organization of VE-cadherin intercellular junctions. Nck determines spatiotemporal patterns of Cdc42/aPKC activation to regulate endothelial apical-basal polarity and lumen formation. Multiple angiogenic cues modulate phosphotyrosine signaling to promote vasculogenesis and angiogenesis. Despite its functional and clinical importance, how vascular cells integrate phosphotyrosine-dependent signaling to elicit cytoskeletal changes required for endothelial morphogenesis remains poorly understood. The family of Nck adaptors couples phosphotyrosine signals with actin dynamics and therefore is well positioned to orchestrate cellular processes required in vascular formation and remodeling. Culture of endothelial cells in three-dimensional collagen matrices in the presence of VEGF stimulation was combined with molecular genetics, optical imaging, and biochemistry to show that Nck-dependent actin remodeling promotes endothelial cell elongation and proper organization of VE-cadherin intercellular junctions. Major morphogenetic defects caused by abrogation of Nck signaling included loss of endothelial apical-basal polarity and impaired lumenization. Time-lapse imaging using a Förster resonance energy transfer biosensor, immunostaining with phospho-specific antibodies, and GST pull-down assays showed that Nck determines spatiotemporal patterns of Cdc42/aPKC activation during endothelial morphogenesis. Our results demonstrate that Nck acts as an important hub integrating angiogenic cues with cytoskeletal changes that enable endothelial apical-basal polarization and lumen formation. These findings point to Nck as an emergent target for effective antiangiogenic therapy.