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: 9102169
• 负责人: George E Davis
• 金额: $ 38.37万
• 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/9102169
• 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; FGF2 gene; Family; Fibroblast Growth Factor; Fibroblast Growth Factor Receptor 2; Galium; Growth; Growth Factor Receptors; Health; Human; In Vitro; Insulin; Interleukin-3; KDR gene; 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-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; 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（由应用程序提供）：在这项新提案中，我们研究了定义新的生长因子组合和信号通路的发现，这些发现控制EC结核病和Ec-Pricyte Tube Tube共同组装，这对于建立毛细管网络是必要的，这些网络非常支持组织灌注，开发，发育，功能维护。毛细血管由两种主要细胞类型组成，即EC和周细胞，它们共组装，形成具有绝对位置周细胞的偏振EC衬管和一个中间的基底膜基质。戴维斯实验室（Davis Lab）使用人类EC和周细胞开发了体外系统的开发，在无血清定义的条件下，这些事件在3D物质中进行了建模。值得注意的是，我们已经确定，添加了五个定义的生长因子，SCF，IL-3，SDF-1A，FGF-2和胰岛素（因子），允许使用参与血管基底膜基质组装的人类EC结核和募集周细胞，这是我们的实验室首次报道的。新的初步数据表明，这些因素及其接收器通过激活三个关键协同信号通路来控制肾变生成。 PI3K/AKT/MTOR，RAF/MEK/ERK和JAK/STAT。这些途径随着时间的推移而被其独有的方式强烈激活和持续，这对于ec tubegeNesis是必不可少的。在VEGF，FGF-2或VEGF+FGF-2添加之后，这种重要的信号传导并未体现。此外，我们确定的系统导致了另一个主要见解，证明了VEGF作为因子诱导的EC tubegenesis和Ec-Pricyte Tube Tube的上游底漆的新作用，同时它无法直接刺激这些过程。我们还报道说，衍生的PDGF-BB和HB-EGF在介导周细胞募集，增殖和基底膜形成方面起着关键作用，这种观察结果在体内证实，使用发育的鹌鹑胚胎在体内证实。最后，我们获得了EC-磨牙管共组装中EC衍生的TGFß1，Activinb，CTGF和CCL2家族趋化因子的作用的新证据。这些新见解共同提供了一个分子路线图，以剖析定义的生长因子，受体，它们的常见适配器以及重叠的信号通路控制EC tubexenesis和Ec-Gricyte Tube Tube Tube Tube Tube Tube均使用体外和体内实验方法共同组装。我们提出了三个特定的目的，以进一步研究这些新颖的见解，以响应定义的生长因子，在体外和体内的毛细管组件的基本过程中进行。 AIM1：识别和表征所需的SCF，IL-3，SDF-1A和FGF-2-以及接收器依赖的信号传导事件，这些事件在体外和体内控制EC结核。 AIM2：为了阐明基本机制如何协同激活信号通路，AKT/MTOR，RAF/ERK和JAK/STAT，导致EC结节发生。 AIM3：阐明SCF，IL-3，SDF-1A和FGF-2-以及受体依赖性信号如何导致Ec-Pricyte Tube Tube聚管和血管基底膜形成。