Molecular mechanism of vascular endothelial/mural cell differentiation by receptor tyrosine kinases

受体酪氨酸激酶分化血管内皮细胞/壁细胞的分子机制

基本信息

批准号：
15570110
负责人：
MIYAZAWA Keiji
金额：
$ 2.37万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

项目摘要

In the present study, we examined the roles of receptor tyrosine kinases in differentiation of mouse embryonic stem (ES) cell-derived VEGFR2^+ cells into endothelial cells and mural cells.(1)VEGFR3 signaling in endothelial differentiationVEGFR3 is a receptor for VEGF-C and VEGF-D. In order to examine the role of VEGFR3 in endothelial differentiation, we established ES cells expressing VEGFR3 by Supertransfection technique and then prepared VEGFR2^+ cells through in vitro differentiation and MACS. These cells differentiated into endothelial cells and expressed lymphatic endothelial marker LYVE-1 in the presence of VEGF-C. VEGF-C stimulates VEGFR3 as well as VEGFR3, whereas VEGF-C (C152S) mutant exclusively stimulates VEGFR3. VEGF-C (C152S) failed to induce endothelial differentiation. Upon stimulation with VEGF-C, VEGFR2 signaling is indispensable for endothelial differentiation, and VEGFR3 signaling additionally confers lymphatic endothelial-like phenotypes to endothelial cells.(2)FGF signaling in endothelial/smooth muscle differentiationWe examined the differentiation of VEGFR2^+ cells in the presence of FGF-2 for 2 days. 45% of the cells differentiated PECAM1^+ endothelial cells, 20% of them differentiated into SMA^+ mural cells, and 35% of them remained negative for both markers. FGF-2 thus does not direct cell differentiation to one specific lineage. We also examined the co-stimulatory effect of VEGF-A and FGF-2. Whereas VEGF-A caused exclusive endothelial differentiation, VEGF-A/FGF-2 co-stimulation induced 95% endothelial cells and 5% mural cells. Notably mural cells attached to the entdothelial sheets. We found that the cell-cell communication was mediated through endogenonus PDGF-B/PDGFRβ signaling.

在本研究中，我们检查了受体酪氨酸激酶在小鼠胚胎干（ES）细胞衍生的VEGFR2^+细胞分化中的作用。（1）vegfr3信号传导在内皮分化中的VEGFR3信号传导是VEGGF-C和VEGGF-D的受体。为了检查VEGFR3在内皮分化中的作用，我们通过超级转染技术建立了ES细胞，然后通过体外分化和MAC制备了VEGFR2^+细胞。这些细胞分化为内皮细胞，并在VEGF-C存在下表达淋巴内皮标记Lyve-1。 VEGF-C刺激VEGFR3和VEGFR3，而VEGF-C（C152S）突变体仅刺激VEGFR3。 VEGF-C（C152S）未能诱导内皮分化。在用VEGF-C刺激后，VEGFR2信号传导对于内皮分化是必不可少的，VEGFR3信号另外将类似淋巴的内皮表型与内皮细胞结合在一起的。 45％的细胞分化了PECAM1^+内皮细胞，其中20％分化为SMA^+壁细胞，其中35％的细胞对这两个标记物仍为阴性。因此，FGF-2不会将细胞分化为一个特定的谱系。我们还研究了VEGF-A和FGF-2的共刺激作用。 VEGF-A引起独家内皮分化，而VEGF-A/FGF-2共刺激诱导了95％的内皮细胞和5％的壁细胞。特别是附着在entdophelial片上的壁细胞。我们发现细胞 - 细胞通信是通过pDGF-B/pDGFRβ信号传导介导的。