The Role of Adenosine as a Modulator of Angiogenesis

腺苷作为血管生成调节剂的作用

• 批准号: 6682342
• 负责人: AMY L TUCKER
• 金额: $ 29.6万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-12-04 至 2005-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6682342
• 关键词: adenosine; angiogenesis; cardiovascular pharmacology; cell growth regulation; chemical kinetics; chickens; chorioallantoic membrane; cytokine; developmental genetics; embryo /fetus cell /tissue; enzyme linked immunosorbent assay; gene expression; genetically modified animals; growth factor; histology; human subject; immunoaffinity chromatography; intermolecular interaction; laboratory mouse; laboratory rat; macrophage; monocyte; protein isoforms; purinergic receptor; species difference

Previous work on the role of adenosine receptor subtypes as pro-angiogenic factors has largely been in vitro and has implicated the A2 receptors on vascular cells including endothelial and smooth muscle cells, as the mediators for the effect. Our in vivo data in the chicken chorioallantoic membrane (CAM) assay unexpectedly indicate that the A1 receptor is among the subtypes that stimulate angiogenesis. Furthermore, we have determined that the A1 adenosine receptor likely works through inflammatory, not vascular, cells by stimulating the release of vascular endothelial growth factor (VEGF) from monocytes/macrophages. We hypothesize that adenosine promotes angiogenesis at a number of different levels. A2 receptor stimulation triggers endothelial cell proliferation and release of VEGF from smooth muscle cells, while A1 receptor stimulation promotes angiogenesis through its effects on inflammatory cells, namely monocytes/macrophages. The A3 receptor may work through effects on mast cells. We have formulated three Specific Aims to test these hypotheses: Aim I): We will use novel subtype-selective ligands and allosteric enhancers of adenosine receptors to pharmacologically characterize the chicken, rat, mouse, and human adenosine receptors. The results of Aim I will guide the selection of ligand concentrations for subsequent aims. Aim II): We will use three models of angiogenesis, the CAM, rat thoracic aortic ring, and rat mesenteric models to demonstrate in vivo modulation of angiogenesis by adenosine agonists, antagonists, and allosteric enhancers. A fourth model, the ischemic rat hindlimb model, is in development and will be used to test the ability of pro-angiogenic ligands and enhancers to improve blood flow to an ischemic muscle bed. Aim III): We will use subtype-selective ligands on cultured cells to identify mechanisms of adenosine-stimulated angiogenesis. We will identify which adenosine receptor subtypes trigger endothelial cell proliferation and which stimulate cytokine release from smooth muscle cells, mast cells, and monocytes/macrophages. Our ultimate goal is therapeutic manipulation of angiogenesis.

先前关于腺苷受体亚型作为促血管生成因子的作用的工作在很大程度上是体外的，并且已将A2受体牵涉到包括内皮和平滑肌细胞在内的血管细胞上，作为该作用的介体。 我们在鸡绒毛膜膜（CAM）测定中的体内数据出乎意料地表明，A1受体是刺激血管生成的亚型之一。 此外，我们已经确定A1腺苷受体可能通过刺激单核细胞/巨噬细胞释放血管内皮生长因子（VEGF）来通过炎症而不是血管来起作用。 我们假设腺苷在多种不同水平上促进血管生成。 A2受体刺激会触发平滑肌细胞的内皮细胞增殖和VEGF释放，而A1受体刺激通过其对炎症细胞的影响（即单核细胞/巨噬细胞）促进血管生成。 A3受体可以通过对肥大细胞的影响来起作用。 我们已经制定了三个特定的目的来检验这些假设：目标i）：我们将使用新型的亚型选择配体和腺苷受体的变构增强剂，以在药理学上表征鸡肉，大鼠，小鼠和人类腺苷受体。 AIM我将指导选择配体浓度以进行后续目标的结果。 AIM II）：我们将使用三种模型的血管生成模型，CAM，大鼠胸主动脉环和大鼠肠系膜模型来证明腺苷激动剂，拮抗剂和变构增强剂对血管生成的体内调节。 第四个模型，缺血性大鼠后肢模型，正在开发中，并将用于测试亲血管生成的配体和增强子的能力，以改善流向缺血性肌肉床的血液流动。 AIM III）：我们将在培养的细胞上使用亚型选择配体来识别腺苷刺激的血管生成的机制。 我们将确定哪种腺苷受体亚型会触发内皮细胞增殖，并刺激细胞因子从平滑肌细胞，肥大细胞和单核细胞/巨噬细胞中释放。 我们的最终目标是对血管生成的治疗操作。