Defining an Upstream VEGF (Vascular Endothelial Growth Factor) Priming Signature for Downstream Factor-Induced Endothelial Cell-Pericyte Tube Network Coassembly.
Defining an Upstream VEGF (Vascular Endothelial Growth Factor) Priming Signature for Downstream Factor-Induced Endothelial Cell-Pericyte Tube Network Coassembly.
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DOI：
10.1161/atvbaha.120.314517
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发表时间：
2020-12
期刊：
影响因子：
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通讯作者：
Davis GE
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文献类型：
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作者：
Bowers SLK;Kemp SS;Aguera KN;Koller GM;Forgy JC;Davis GE
在这项工作中，我们试图确定生长因子的需求和信号基础的不同阶段的人血管形态发生和成熟。使用依赖于SCF、IL-3、SDF-1α、FGF-2和胰岛素（因子）五种生长因子组合的3D胶原基质中EC管形态发生的无血清模型，我们证明了VEGF预处理EC 8小时（即VEGF引发）导致EC对因子的应答显著增加，所述因子包括：EC尖端细胞、EC小管发生、周细胞募集和增殖以及基底膜沉积。VEGF致敏需要VEGFR 2，VEGFR 2的作用对致敏反应具有选择性，并且在不存在致敏的情况下不影响因子依赖性小管发生。VEGF引发的关键分子和信号传导要求包括RhoA、Rock 1、PKCα和PKD 2。这些分子和信号传导途径的siRNA抑制或药理学阻断干扰VEGF作为下游因子依赖性EC管形成以及周细胞募集的上游引物的能力。VEGF引发还与肌动蛋白应力纤维的形成、粘着斑成分的活化、EC因子受体、c-Kit、IL-3Rα和CXCR 4的上调以及EC衍生的PDGF-BB、PDGF-DD和HB-EGF的上调相关，这些共同影响周细胞募集和增殖。总的来说，这项研究定义了一个可分离的上游VEGF引发步骤的信号传导特征，该步骤可以激活EC以响应下游因子，这些下游因子是形成具有相关壁细胞的分支管网所必需的。
In this work, we have sought to define growth factor requirements and the signaling basis for different stages of human vascular morphogenesis and maturation. Using a serum-free model of EC tube morphogenesis in 3D collagen matrices that depends on a five growth factor combination, SCF, IL-3, SDF-1α, FGF-2, and insulin (Factors), we demonstrate that VEGF pre-treatment of ECs for 8 hr (i.e. VEGF priming) leads to marked increases in the EC response to the Factors which includes; EC tip cells, EC tubulogenesis, pericyte recruitment and proliferation, and basement membrane deposition. VEGF priming requires VEGFR2 and the effect of VEGFR2 is selective to the priming response and does not affect Factor-dependent tubulogenesis in the absence of priming. Key molecule and signaling requirements for VEGF priming include RhoA, Rock1, PKCα, and PKD2. siRNA suppression or pharmacologic blockade of these molecules and signaling pathways interfere with the ability of VEGF to act as an upstream primer of downstream Factor-dependent EC tube formation as well as pericyte recruitment. VEGF priming was also associated with the formation of actin stress fibers, activation of focal adhesion components, upregulation of the EC Factor receptors, c-Kit, IL-3Rα, and CXCR4, and upregulation of EC-derived PDGF-BB, PDGF-DD, and HB-EGF which collectively affect pericyte recruitment and proliferation. Overall, this study defines a signaling signature for a separable upstream VEGF priming step, which can activate ECs to respond to downstream Factors that are necessary to form branching tube networks with associated mural cells.