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（申请人提供）：在这项新的提案中，我们研究了我们的新发现，造血干细胞细胞因子的独特组合（即干细胞因子-SCF;白细胞介素3- IL-3;和基质衍生因子-1 α- SDF-11）在限定的无血清条件下控制人内皮细胞（EC）管形态发生和EC-周细胞管共组装（并且在不存在佛波醇酯的情况下）在3D胶原基质中的组合物。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发芽产生影响，这是其主要的形态发生影响。我们提出的假设是，SCF，IL-3和SDF-11是血管形态发生的关键调节因子，它们以协同的方式起作用，并且是已知的发育血管形成介质（例如VEGF和BMP-4）的作用的必要辅助因子。联合施用c-Kit（SCF受体）和CXCR 4（SDF-11受体）抑制剂或SCF和IL-3的阻断抗体导致发育中的鹌鹑胚胎中的血管出血表型，其与血管形成和重塑中的缺陷有关。此外，我们表明，这三个因素协同激活Src，B-Raf和Erk激酶，并诱导1221整合素和Pak 2的表达，这是已知的调节血管形态发生。我们将阐明三种细胞因子在EC中的信号协同作用的分子基础，这些因子如何与VEGF，BMP-4和周细胞相互作用以控制EC发芽，以及它们如何在体外和体内影响EC和周细胞在管组装和成熟过程中的作用。 我们提出了三个具体目标：具体目标#1。阐明SCF、IL-3和SDF-11协同刺激血管生成EC管在三维细胞外基质中组装的信号机制。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EC在3D细胞外基质中的发芽反应。具体目标#3确定SCF、IL-3和SDF-11如何调节周细胞诱导的EC出芽反应和血管生成EC-周细胞管在三维细胞外基质中的共组装。 公共卫生关系：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。这些因子与内皮细胞结合，内皮细胞排列在血管和周细胞上，周细胞是一种帮助血管形成和稳定的支持细胞。对血管如何形成和稳定的基本机制的基本理解对于在各种人类疾病如心血管疾病、糖尿病或癌症的背景下刺激或抑制该过程的努力至关重要。