Investigation of the role of Eph/Ephrin signaling in vasculogenesis and angiogenesis

Eph/Ephrin 信号在血管发生和血管生成中的作用的研究

• 批准号: 12835011
• 负责人: OIKE Yuichi
• 金额: $ 2.37万
• 依托单位: Kumamoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12835011/
• 关键词: Vasculogenesis; Angiogenesis; Eph; Ephrin Signaling; 血管生物学; 再生医学; 分子生物学; 受容体型チロシンキナーゼ

Although the cellular and molecular mechanisms governing angiogenesis are only beginning to be understood, signaling through endothelial-restricted receptors, particularly receptor tyrosine kinases, have been shown to play a pivotal role in these events. Recent reports show that EphB receptor tyrosine kinases and their transmembrane-type ephrin-B2 ligands play essential roles in the embryonic vasculature. Eph receptor tyrosine kinases and their cell-surface-bound ligands, the ephrins mediate cell migration and adhesion. They contribute to developmental vascular assembly and body patterning, including directing axon guidance and building in the developing brain. These suggest that cell-to-cell repellent effects due to bi directional EphB/ephrin-B2 signaling may be crucial for vascular development similar to the mechanism described for neuronal development. To test this hypothesis, we disrupted the precise expression pattern of EphB/ephrin-B2 in vivo by generating transgenic (CAGp-ephrin … More -B2 Tg) mice that express ephrin-B2 under the control of a ubiquitous and constitutive promoter, CAG. These mice displayed an abnormal segmental arrangement of intersomitic vessels, while such anomalies were not observed in Tie-2p-ephrin-B2 Tg mice in which ephrin-B2 was overexpressed in only vascular endothelial cells (ECs). This finding suggests that non ECs expressing ephrin-B2 alter the migration of ECs expressing EphB receptors into the intersomitic region where ephrin-B2 expression is normally absent. CAGp-ephrin-B2 Tg mice show sudden death at neonatal stages from aortic dissecting aneurysms due to defective recruitment of vascular smooth muscle cells to the ascending aorta. Next, we investigate the role of ephrinB2 on mesenchymal cells in vascular development. To do so, we analyzed the vascular bed and network formation by para-aortic splanchnopleural mesoderm (P-Sp) explants which are rich of endothelial precursor cells from mouse embryos (E9.5) using a co-culture system with OP9 stromal cells transfected with vector or ephrin B2. Interestingly, OP9/ephrinB2 stromal cells promote both proliferation and sprouting/remodeling of ephrinB2-positive endothelial cells and smooth muscle actin-positive pericytes. These findings demonstrate that EphB/ephrin-B2 signaling between endothelial cells and surrounding mesenchymal cells plays an essential role in vasculogenesis, angiogenesis and vessel maturation. As vessel projection is initial event in many pathological condition, it may provide new strategies in (anti)-angiogenic treatment. Less

虽然控制血管生成的细胞和分子机制才刚刚开始被理解，通过内皮限制性受体，特别是受体酪氨酸激酶的信号转导，已被证明在这些事件中发挥关键作用。最近的报道表明，EphB受体酪氨酸激酶和它们的跨膜型ephrin-B2配体在胚胎血管系统中发挥重要作用。Eph受体酪氨酸激酶及其细胞表面结合配体肝配蛋白介导细胞迁移和粘附。它们有助于发育中的血管组装和身体模式，包括指导轴突引导和在发育中的大脑中构建。这些表明，由于双向EphB/肝配蛋白-B2信号传导引起的细胞对细胞的排斥作用可能对血管发育至关重要，类似于神经元发育所描述的机制。为了验证这一假设，我们通过产生转基因（CAGp-ephrin）来破坏EphB/ephrin-B2在体内的精确表达模式 ...更多信息 -B2 Tg）小鼠，其在普遍存在的组成型启动子CAG的控制下表达肝配蛋白-B2。这些小鼠显示异常的体节间血管节段性排列，而在Tie-2 p-ephrin-B2 Tg小鼠中未观察到此类异常，其中ephrin-B2仅在血管内皮细胞（EC）中过表达。这一发现表明，不表达肝配蛋白-B2的内皮细胞改变表达EphB受体的内皮细胞迁移到通常不表达肝配蛋白-B2的体间区。CAGp-ephrin-B2 Tg小鼠在新生期由于血管平滑肌细胞向升主动脉的募集缺陷而死于主动脉夹层动脉瘤。接下来，我们研究ephrinB 2在血管发育中对间充质细胞的作用。为此，我们分析了血管床和网络形成的腹主动脉旁内脏胸膜中胚层（P-Sp）外植体，这是丰富的内皮前体细胞从小鼠胚胎（E9.5）使用共培养系统与OP 9基质细胞转染载体或肝配蛋白B2。有趣的是，OP 9/ephrinB 2基质细胞促进ephrinB 2阳性内皮细胞和平滑肌肌动蛋白阳性周细胞的增殖和发芽/重塑。这些发现表明内皮细胞和周围间充质细胞之间的EphB/ephrin-B2信号传导在血管发生、血管生成和血管成熟中起重要作用。由于血管投射是许多病理状态下的初始事件，它可能为（抗）血管生成治疗提供新的策略。少

项目成果

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