The neuronal transcription factor NPAS4 is a strong inducer of sprouting angiogenesis and tip cell formation
The neuronal transcription factor NPAS4 is a strong inducer of sprouting angiogenesis and tip cell formation
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DOI：
10.1093/cvr/cvw248
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发表时间：
2017-02-01
影响因子：
10.8
通讯作者：
Moser, Martin
中科院分区：
文献类型：
--
作者：
Esser, Jennifer Susanne;Charlet, Anne;Moser, Martin
基本原理关于分支形态发生，神经发生和血管生成具有共同的原理机制并利用相同的分子。因此，参与血管形态发生的神经元分子的调查提供了新的可能性，在cardiovascular diseases.Objective促血管生成的方法在这项研究中，我们调查了神经元转录因子NPAS 4在angiogenesis.Methods and results中的作用在这里，我们证明了神经元转录因子NPAS 4在不同来源的内皮细胞中表达，使用反转录PCR和Western blot分析。为了研究NPAS 4如何影响内皮细胞功能，NPAS 4通过质粒转染过表达或通过特异性siRNA从人脐静脉内皮细胞（HUVECs）中去除。体外HUVEC发芽试验表明，NPAS 4过表达增强了内皮细胞的发芽和分支。因此，NPAS 4的沉默导致HUVEC发芽和分支减少。从机制上讲，我们确定了靶基因血管内皮粘附分子VE-钙粘蛋白参与NPAS 4的促血管生成功能。在内皮细胞镶嵌球体发芽试验中，NPAS 4参与尖端细胞的形成。小鼠和斑马鱼的体内实验证实了我们的体外研究结果。NPAS 4缺陷小鼠在基质胶塞测定中显示内皮细胞向内生长减少。与NPAS 4在内皮细胞功能中的调节作用一致，通过特异性morpholinos沉默斑马鱼中的NPAS 4导致了段间血管生长的干扰。结论NPAS 4在内皮细胞中表达，调节VE-钙粘蛋白的表达，并调节出芽血管生成。
Rationale Regarding branching morphogenesis, neurogenesis and angiogenesis share common principle mechanisms and make use of the same molecules. Therefore, the investigation of neuronal molecules involved in vascular morphogenesis provides new possibilities for pro-angiogenic approaches in cardiovascular diseases.Objective In this study, we investigated the role of the neuronal transcription factor NPAS4 in angiogenesis.Methods and results Here, we demonstrate that the neuronal transcription factor NPAS4 is expressed in endothelial cells of different origin using reverse transcription PCR and western blot analysis. To investigate how NPAS4 affects endothelial cell function, NPAS4 was overexpressed by plasmid transfection or depleted from human umbilical vein endothelial cells (HUVECs) by specific siRNAs. In vitro HUVEC sprouting assays showed that sprouting and branching of endothelial cells was enhanced by NPAS4 overexpression. Consistently, silencing of NPAS4 resulted in reduced HUVEC sprouting and branching. Mechanistically, we identified as target gene vascular endothelial adhesion molecule VE-cadherin to be involved in the pro-angiogenic function of NPAS4. In endothelial cell mosaic spheroid sprouting assays, NPAS4 was involved in tip cell formation. In vivo experiments in mouse and zebrafish confirmed our in vitro findings. NPAS4-deficient mice displayed reduced ingrowth of endothelial cells in the Matrigel plug assay. Consistent with a regulatory role of NPAS4 in endothelial cell function silencing of NPAS4 in zebrafish by specific morpholinos resulted in perturbed intersegmental vessels growth.Conclusions NPAS4 is expressed in endothelial cells, regulates VE-cadherin expression and regulates sprouting angiogenesis.