Hematopoietic stem cell cytokine control of developmental vascularization

造血干细胞细胞因子控制发育血管化

• 批准号: 8402619
• 负责人: George E Davis
• 金额: $ 36.06万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-03 至 2014-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8402619
• 关键词: Abdomen; Address; Affect; Angiopoietins; Aorta; Basement membrane; Binding; Biological Assay; Blocking Antibodies; Blood Cells; Blood Vessels; Brain; CD80 gene; CXCR4 gene; Cardiovascular Diseases; Cell Line; Cell Proliferation; Cell Survival; Collagen; Complex; Cytokine Receptors; Data; Defect; Development; Developmental Process; Diabetes Mellitus; Dorsal; Embryo; Endothelial Cells; Event; Extracellular Matrix; Fibrin; Growth; Growth Factor; Hematopoiesis; Hematopoietic; Hematopoietic stem cells; Hemorrhage; Human; In Vitro; Integrins; Interleukin-3; Laboratories; Ligands; Link; Malignant Neoplasms; Mediator of activation protein; Mesenchyme; Modeling; Molecular; Morphogenesis; Pericytes; Phenotype; Phorbol Esters; Phosphotransferases; Platelet-Derived Growth Factor; Process; Protein Isoforms; Proto-Oncogene Protein c-kit; Quail; Serum; Serum-Free Culture Media; Signal Transduction; Stem Cell Factor; Stromal Cell-Derived Factor 1; Supporting Cell; System; Transforming Growth Factor beta; Tube; Vascular Endothelial Growth Factors; Vascularization; Work; base; bone morphogenic protein; cell behavior; cofactor; cytokine; fascinate; human disease; in vivo; inhibitor/antagonist; interest; monolayer; novel; precursor cell; receptor; response; screening; stem; synergism; vasculogenesis

In this new proposal, we investigate our novel finding that a unique combination of hematopoietic stem cell cytokines (i.e. stem cell factor-SCF; interleukin 3- IL-3; and stromal-derived factor-1 alpha- SDF-1¿) control human endothelial cell (EC) tube morphogenesis and EC-pericyte tube coassembly under defined serum-free conditions (and in the absence of phorbol ester) in 3D collagen matrices. Interestingly, VEGF has no influence by itself and requires the synergistic action of these hematopoietic factors in order to exert an effect on EC sprouting, its major morphogenic influence. We propose the hypothesis that SCF, IL-3 and SDF-1¿ are critical regulators of vascular morphogenesis which act in a synergistic manner and are necessary cofactors for the action of known mediators of developmental vascularization (e.g. VEGF and BMP-4). Combined administration of c-Kit (the SCF receptor) and CXCR4 (the SDF-1¿ receptor) inhibitors or blocking antibodies to SCF and IL-3 leads to vascular hemorrhage phenotypes in developing quail embryos that relates to defects in vessel formation and remodeling. Also, we show that the three factors synergize to activate Src, B-Raf and Erk kinases and induce the expression of the ¿2¿1 integrin and Pak2 which are known regulators of vascular morphogenesis. We will elucidate the molecular basis for the signaling synergism of the three cytokines in ECs, how these factors interface with VEGF, BMP-4, and pericytes to control EC sprouting and how they affect both ECs and pericytes during tube coassembly and maturation events in vitro and in vivo. We propose three specific aims which are; Specific Aim #1. To elucidate the signaling mechanisms by which SCF, IL-3 and SDF-1¿ synergize to stimulate vasculogenic EC tube assembly in 3D extracellular matrices. Specific Aim #2. To determine how SCF, IL-3 and SDF-1¿ affect VEGF and BMP-4 signaling to control EC sprouting responses in 3D extracellular matrices. Specific Aim #3. To determine how SCF, IL-3 and SDF-1¿ act to regulate pericyte-induced EC sprouting responses and vasculogenic EC-pericyte tube coassembly in 3D extracellular matrices.

在这个新提案中，我们调查了我们的小说发现，即造血茎的独特组合 细胞细胞因子（即干细胞因子-SCF；白细胞介素3- IL-3；以及基质衍生的因子-1 alpha-SDF-1） 控制人内皮细胞（EC）管形形态发生和Ec-Pricyte Tube管结合下 在3D胶原蛋白材料中定义了无血清条件（并且在没有佛波酯的情况下）。 有趣的是，VEGF本身没有影响，需要这些造血因素的协同作用 为了对EC发芽产生影响，其主要的形态学影响。我们提出了以下假设 SCF，IL-3和SDF-1…是血管形态发生的关键调节剂，以协同的方式起作用，并且 对于已知的发育血管化介体的作用是必要的辅助因子（例如VEGF和 BMP-4）。 C-KIT（SCF受体）和CXCR4（SDF-1听受体）抑制剂的联合给药或 阻断对SCF和IL-3的抗体导致发育中的鹌鹑胚胎的血管出血表型 这与血管形成和重塑的缺陷有关。另外，我们表明这三个因素与 激活SRC，B-RAF和ERK激酶，并影响已知的2¿1整合蛋白和PAK2的表达 血管形态发生的调节剂。我们将阐明分子基础的信号传导协同作用 EC中的三种细胞因子，这些因素如何与VEGF，BMP-4和周细胞接口以控制EC发芽 以及它们如何在体外和体内影响管结合和成熟事件期间的EC和周细胞。 我们提出了三个特定目标： 特定目标＃1。为了阐明SCF，IL-3和SDF-1的信号传导机制协同为 刺激3D细胞外物质中的血管生成EC管组件。 特定目标＃2。为了确定SCF，IL-3和SDF-1。如何影响VEGF和BMP-4信号传导 EC在3D细胞外材料中发芽反应。 特定目标＃3。确定SCF，IL-3和SDF-1€如何调节周细胞周围的EC 在3D细胞外材料中的发芽反应和血管生成的Ec-Pricyte管结合。