Novel growth factor and signaling requirements for human capillary tube assembly

人体毛细管组装的新型生长因子和信号传导要求

• 批准号: 8942261
• 负责人: George E Davis
• 金额: $ 39.7万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8942261
• 关键词: Address; Affect; Automobile Driving; Basement membrane; Biological Assay; Blood; Blood Vessels; Blood capillaries; CCL2 gene; Cardiovascular Diseases; Cell Culture Techniques; Cell Line; Cells; Coupling; DTR gene; Data; Deposition; Development; Diabetes Mellitus; Disease; Embryo; Endothelial Cells; Event; Exogenous Factors; Family; Fibroblast Growth Factor; Fibroblast Growth Factor 2; Fibroblast Growth Factor Receptor 2; Galium; Growth Factor; Growth Factor Receptors; Human; In Vitro; Insulin; Interleukin-3; Laboratories; Lead; Maintenance; Malignant Neoplasms; Maps; Mediating; Membrane Protein Traffic; Modeling; Molecular; Monomeric GTP-Binding Proteins; Morphogenesis; Mus; Pathway interactions; Perfusion; Pericytes; Phosphotransferases; Play; Positioning Attribute; Process; Quail; Recruitment Activity; Reporting; Role; Serum; Serum-Free Culture Media; Signal Pathway; Signal Transduction; Signaling Molecule; Small Interfering RNA; Stem Cell Factor; System; Time Factors; Tissues; Tube; Vascular Endothelial Growth Factor Receptor-2; Vascular Endothelial Growth Factor Receptor-3; Vascular Endothelial Growth Factors; Work; blood vessel development; capillary; cell type; chemokine; connective tissue growth factor; in vivo; insight; member; membrane assembly; novel; novel strategies; platelet-derived growth factor BB; public health relevance; receptor; response

 DESCRIPTION (provided by applicant): In this new proposal, we investigate our findings defining novel growth factor combinations and signaling pathways that control EC tubulogenesis and EC-pericyte tube co-assembly, which are necessary to create capillary networks, which critically support tissue perfusion, development and functional maintenance. Capillaries consist of two major cell types, ECs and pericytes, which co-assemble to form polarized EC-lined tubes with abluminally positioned pericytes and an intervening basement membrane matrix. The Davis lab has pioneered the development of in vitro systems using human ECs and pericytes, which model these events in 3D matrices under serum-free defined conditions. Remarkably, we have identified that addition of five defined growth factors, SCF, IL-3, SDF-1a, FGF-2, and insulin (Factors), allows for human EC tubulogenesis and recruitment of pericytes with accompanying vascular basement membrane matrix assembly, a finding first reported by our laboratory. Novel preliminary data reveals that these Factors and their receptors control tubulogenesis through activation of three key synergistic signaling pathways; PI3K/Akt/mTor, Raf/Mek/Erk, and Jak/Stat. These pathways are strongly activated and sustained over time by the Factors in a manner that is unique to them, and which are necessary for EC tubulogenesis; and this important signaling is not manifest following VEGF, FGF-2 or VEGF+FGF-2 addition. Furthermore, our defined system led to another major insight demonstrating a novel role for VEGF as an upstream primer of Factor-induced EC tubulogenesis and EC-pericyte tube co-assembly, while it fails to stimulate these processes directly. We also reported that EC- derived PDGF-BB and HB-EGF play a key role in mediating pericyte recruitment, proliferation and basement membrane formation, an observation that was confirmed in vivo using developing quail embryos. Finally, we have obtained novel evidence for a role for EC-derived TGFß1, activinB, CTGF, and CCL2 family chemokines in EC-pericyte tube co-assembly. Together, these new insights provide a molecular road map to dissect how defined growth factors, receptors, their common adapters, and overlapping signaling pathways control EC tubulogenesis and EC-pericyte tube co-assembly using both in vitro and in vivo experimental approaches. We propose three specific aims to further investigate these novel insights into the fundamental process of capillary tube assembly in vitro and in vivo in response to defined growth factors and they are; Aim1: To identify and characterize required SCF, IL-3, SDF-1a and FGF-2- and receptor-dependent signaling events that control EC tubulogenesis in vitro and in vivo. Aim2: To elucidate the underlying mechanisms how Factor receptors and adapters synergistically work together to activate the signaling pathways, Akt/mTor, Raf/Erk, and Jak/Stat, leading to EC tubulogenesis. Aim3: To elucidate how SCF, IL-3, SDF-1a and FGF-2- and receptor-dependent signaling leads to EC- pericyte tube co-assembly and vascular basement membrane formation.

 描述（由申请人提供）：在这项新提案中，我们研究了我们的发现，定义了控制 EC 管发生和 EC 周细胞管共组装的新型生长因子组合和信号通路，这些是创建毛细血管网络所必需的，毛细血管网络对组织灌注至关重要毛细血管由两种主要细胞类型组成，即 EC 和周细胞，它们共同组装形成位于管腔外的极化 EC 衬里管。戴维斯实验室率先开发了使用人类 EC 和周细胞的体外系统，该系统在无血清限定条件下在 3D 矩阵中模拟了这些事件。值得注意的是，我们已经确定了五个确定的生长的添加。因子、SCF、IL-3、SDF-1a、FGF-2 和胰岛素（因子），允许人类 EC 管发生和募集周细胞以及伴随的血管基底膜基质我们实验室首次报道的一项新发现表明，这些因子及其受体通过激活 PI3K/Akt/mTor、Raf/Mek/Erk 和 Jak/Stat 三个关键的协同信号通路来控制肾小管发生。随着时间的推移，这些因子以其独​​特的方式激活和维持，这对于 EC 肾小管发生是必需的，并且这种重要的信号传导在 VEGF、FGF-2 或此外，我们定义的系统产生了另一个重要见解，证明 VEGF 作为因子诱导的 EC 管发生和 EC 周细胞管共组装的上游引物的新作用，但它无法直接刺激这些过程。我们还报道了 EC 衍生的 PDGF-BB 和 HB-EGF 在介导周细胞募集、增殖和基底膜形成中发挥着关键作用，这一观察结果在体内发育中的鹌鹑胚胎中得到了证实。我们已经获得了 EC 衍生的 TGFβ1、activinB、CTGF 和 CCL2 家族趋化因子在 EC 周细胞管共组装中的作用的新证据，这些新见解为剖析如何定义生长因子、受体、我们使用体外和体内实验方法来进一步研究它们的共同适配器和重叠信号通路控制 EC 管发生和 EC 周细胞管共组装。这些对体外和体内毛细管组装响应特定生长因子的基本过程的新颖见解是：识别和表征所需的 SCF、IL-3、SDF-1a 和 FGF-2- 以及受体-体外和体内控制 EC 管发生的依赖性信号事件 目的2：阐明因子受体和接头如何协同作用以激活信号通路 Akt/mTor 的潜在机制。 Raf/Erk 和 Jak/Stat，导致 EC 管发生 目标 3：阐明 SCF、IL-3、SDF-1a 和 FGF-2 以及受体依赖性信号传导如何导致 EC 周细胞管共组装和血管基底。膜的形成。