Adrenomedullin/cyclic AMP pathway induces notch activation and differentiation of arterial endothelial cells from vascular progenitors
Adrenomedullin/cyclic AMP pathway induces notch activation and differentiation of arterial endothelial cells from vascular progenitors
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DOI：
10.1161/01.atv.0000234978.10658.41
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发表时间：
2006-09-01
影响因子：
8.7
通讯作者：
Yamashita, Jun K.
中科院分区：
文献类型：
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作者：
Yurugi-Kobayashi, Takami;Itoh, Hiroshi;Yamashita, Jun K.
目的：动脉或静脉特性的获得在血管发育中被强调。此前，我们已经报道了一种胚胎干细胞分化系统，该系统使用血管内皮生长因子受体2(VEGFR2)阳性细胞作为常见的血管前体细胞，显示出早期的血管发育。在本研究中，我们在体外对动脉和静脉内皮细胞(ECs)进行了建设性的诱导分化，以阐明动静脉分化的分子机制。方法和结果-从VEGFR2(+)祖细胞在不同的条件下诱导ECs。血管内皮细胞生长因子是诱导内皮细胞的必需因子。8溴cAMP或肾上腺髓质素(AM)是一种内源性配体升高的cAMP，可促进血管内皮生长因子诱导的EC分化。而单独使用血管内皮生长因子主要诱导静脉内皮细胞，8溴-环磷酸腺苷(或AM)与血管内皮生长因子联合使用可显著促进动脉内皮细胞的诱导。刺激cAMP通路可诱导内皮细胞Notch信号的激活。在缺乏Notch信号激活的J kappa位点缺陷的ES细胞中，重组识别序列结合蛋白中的血管内皮生长因子和cAMP的动脉化作用被取消，或者在用γ-分泌酶抑制剂处理的ES细胞中。结论肾上腺髓质素/cAMP是一种新的信号通路，在内皮细胞分化过程中激活Notch信号是一种新的途径。血管内皮生长因子、Notch和cAMP信号的协同作用是诱导血管祖细胞分化为血管内皮细胞所必需的。
Objective - The acquisition of arterial or venous identity is highlighted in vascular development. Previously, we have reported an embryonic stem ( ES) cell differentiation system that exhibits early vascular development using vascular endothelial growth factor ( VEGF) receptor-2 ( VEGFR2)- positive cells as common vascular progenitors. In this study, we constructively induced differentiation of arterial and venous endothelial cells ( ECs) in vitro to elucidate molecular mechanisms of arterial- venous specification.Methods and Results - ECs were induced from VEGFR2(+) progenitor cells with various conditions. VEGF was essential to induce ECs. Addition of 8bromo-cAMP or adrenomedullin ( AM), an endogenous ligand-elevating cAMP, enhanced VEGF-induced EC differentiation. Whereas VEGF alone mainly induced venous ECs, 8bromo-cAMP ( or AM) with VEGF supported substantial induction of arterial ECs. Stimulation of cAMP pathway induced Notch signal activation in ECs. The arterializing effect of VEGF and cAMP was abolished in recombination recognition sequence binding protein at the J kappa site deficient ES cells lacking Notch signal activation or in ES cells treated with gamma-secretase inhibitor. Nevertheless, forced Notch activation by the constitutively active Notch1 alone did not induce arterial ECs.Conclusions - Adrenomedullin/cAMP is a novel signaling pathway to activate Notch signaling in differentiating ECs. Coordinated signaling of VEGF, Notch, and cAMP is required to induce arterial ECs from vascular progenitors.