Functional analysis of the Hey bHLH factors in vascular development

Hey bHLH因子在血管发育中的功能分析

• 批准号: 94041408
• 负责人: Professor Dr. Manfred Gessler
• 金额: --
• 依托单位: Lehrstuhl für Entwicklungsbiochemie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2009
• 资助国家: 德国
• 起止时间: 2008-12-31 至 无数据
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/94041408?language=en
• 关键词: Functional; analysis; Hey; bHLH; factors

The Notch signalling pathway with Hey proteins as bHLH transducers is now firmly established as a key signalling system for embryonic cardiovascular development and adult pathologies in vertebrates. We and others have shown that these genes are essential for establishing arterial identity, suppression of venous fate and angiogenic sprouting and remodelling of developing arteries. Hey1/2 also modulate the cellular hypoxia response. However, there is little insight into the mechanisms whereby Hey proteins influence differentiation and accomplish cell fate changes. We will utilize mouse ES cell differentiation with genetic selection to generate embryonic endothelia in 2D and 3D culture conditions. Expression profiling of wild type and Heyless mutant cells will identify potential downstream mediators of angiogenic fate decisions. In parallel we will expand our whole genome chromatin IP analyses (ChIPseq) to define the full complement of binding sites for Hey proteins in endothelial cells. This will provide us with a systems biology view of Hey targets that we will further validate using forced expression/RNAi and our knockout mice. One focus will be to decipher the reciprocal antagonistic action of Hey and Coup-TFII in artery/vein decisions. We expect these data to greatly enhance our understanding of endothelial differentiation, arterial fate decision, and tip/stalk cell communication in angiogenic sprouting.

以Hey蛋白作为bHLH转换器的Notch信号通路现已牢固确立为脊椎动物胚胎心血管发育和成年病理学的关键信号系统。我们和其他人已经表明，这些基因是建立动脉身份，抑制静脉命运和血管生成发芽和重塑发展中的动脉必不可少的。Hey 1/2还调节细胞缺氧反应。然而，很少有深入了解的机制，使Hey蛋白影响分化和完成细胞命运的变化。 我们将利用小鼠ES细胞分化和遗传选择，在二维和三维培养条件下产生胚胎内皮细胞。野生型和Heyless突变体细胞的表达谱将鉴定血管生成命运决定的潜在下游介质。同时，我们将扩大我们的全基因组染色质IP分析（ChIPseq），以确定内皮细胞中Hey蛋白结合位点的完整补充。这将为我们提供Hey靶点的系统生物学观点，我们将使用强制表达/RNAi和我们的敲除小鼠进一步验证。重点之一是破译Hey和Coup-TFII在动脉/静脉决策中的相互对抗作用。我们希望这些数据能大大增强我们对血管生成萌芽中内皮分化、动脉命运决定和尖端/柄细胞通讯的理解。