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: 8021934
• 负责人: George E Davis
• 金额: $ 37.88万
• 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/8021934
• 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; Screening procedure; 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; stem; synergism; vasculogenesis

DESCRIPTION (provided by applicant): 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-11) control human endothelial cell (EC) tube morphogenesis and EC-pericyte tube co assembly 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-11 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-11 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 1221 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 co assembly 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-11 synergize to stimulate vasculogenic EC tube assembly in 3D extracellular matrices. Specific Aim #2. To determine how SCF, IL-3 and SDF-11 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-11 act to regulate pericyte-induced EC sprouting responses and vasculogenic EC-pericyte tube co assembly in 3D extracellular matrices. PUBLIC HEALTH RELEVANCE: This work focuses on the ability of factors which are known to support the growth of immature blood cells to also be able to support the formation of blood vessels which carry blood cells. These factors bind to endothelial cells, which line blood vessels and to pericytes, a supporting cell that helps blood vessels form and become stable. A basic understanding of the mechanisms underlying how blood vessels form and become stabilized is critical in efforts to stimulate or inhibit the process in the context of various human diseases such as cardiovascular disease, diabetes or cancer.

描述（由申请人提供）：在这项新提案中，我们研究了我们的新发现，即造血干细胞细胞因子（即干细胞因子-SCF、白细胞介素 3-IL-3 和基质衍生因子 1 α-）的独特组合SDF-11) 在规定的无血清条件下（且不存在佛波酯）控制人内皮细胞 (EC) 管形态发生和 EC-周细胞管共组装3D 胶原蛋白基质。有趣的是，VEGF 本身没有影响，需要这些造血因子的协同作用才能对 EC 出芽产生影响，这是其主要的形态发生影响。我们提出这样的假设：SCF、IL-3 和 SDF-11 是血管形态发生的关键调节因子，它们以协同方式发挥作用，并且是已知的发育血管化介质（例如 VEGF 和 BMP-4）作用的必要辅助因子。联合使用 c-Kit（SCF 受体）和 CXCR4（SDF-11 受体）抑制剂或 SCF 和 IL-3 阻断抗体会导致发育中鹌鹑胚胎出现血管出血表型，这与血管形成和重塑缺陷有关。此外，我们还表明，这三个因子协同激活 Src、B-Raf 和 Erk 激酶，并诱导 1221 整合素和 Pak2 的表达，它们是已知的血管形态发生调节因子。我们将阐明 EC 中三种细胞因子信号协同作用的分子基础，这些因子如何与 VEGF、BMP-4 和周细胞相互作用来控制 EC 萌芽，以及它们如何在管共组装和成熟事件中影响 EC 和周细胞。体外和体内。 我们提出三个具体目标：具体目标#1。阐明 SCF、IL-3 和 SDF-11 协同刺激 3D 细胞外基质中血管生成 EC 管组装的信号传导机制。具体目标#2。确定 SCF、IL-3 和 SDF-11 如何影响 VEGF 和 BMP-4 信号传导以控制 3D 细胞外基质中的 EC 出芽反应。具体目标#3。确定 SCF、IL-3 和 SDF-11 如何发挥作用来调节 3D 细胞外基质中周细胞诱导的 EC 出芽反应和血管生成 EC-周细胞管共组装。 公共健康相关性：这项工作的重点是已知支持未成熟血细胞生长的因子也能够支持携带血细胞的血管形成的能力。这些因子与排列血管的内皮细胞和周细胞（一种帮助血管形成并变得稳定的支持细胞）结合。对血管形成和稳定的机制的基本了解对于在心血管疾病、糖尿病或癌症等各种人类疾病的背景下刺激或抑制这一过程至关重要。