IGF2 and IGF1R identified as novel tip cell genes in primary microvascular endothelial cell monolayers.
IGF2 and IGF1R identified as novel tip cell genes in primary microvascular endothelial cell monolayers.
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DOI：
10.1007/s10456-018-9627-4
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发表时间：
2018-11
期刊：
影响因子：
9.8
通讯作者：
Schlingemann RO
中科院分区：
文献类型：
--
作者：
Dallinga MG;Yetkin-Arik B;Kayser RP;Vogels IMC;Nowak-Sliwinska P;Griffioen AW;van Noorden CJF;Klaassen I;Schlingemann RO
使用 CD34 作为标记物，在人脐静脉内皮细胞 (HUVEC) 培养物中鉴定出尖端细胞，即血管生成芽的主导细胞。在这里，我们发现尖端细胞也存在于原代人微血管内皮细胞（hMVEC）中，这是一种与血管生成更相关的内皮细胞类型。通过流式细胞术、免疫细胞化学和 qPCR，显示内皮细胞培养物含有动态的 CD34+ 细胞群，具有尖端细胞的许多特征，包括丝状伪足样延伸、已知尖端细胞基因的 mRNA 水平升高以及对 VEGF 刺激和 DLL4 抑制的反应。此外，我们证明我们的体外尖端细胞模型可用于研究尖端细胞的细胞和分子机制，并发现患者抗血管生成治疗的新靶点。使用小干扰 RNA (siRNA) 敲低已知尖端细胞基因血管生成素 2 (ANGPT2) 和具有免疫球蛋白样和 EGF 样结构域 1 (TIE1) 的酪氨酸激酶的基因表达，与体内抑制尖端细胞相比，这对尖端细胞和出芽产生类似的影响。最后，我们在体外鉴定了 CD34+ 尖端细胞中的两个新的尖端细胞特异性基因：胰岛素样生长因子 2 (IGF2) 和 IGF-1 受体 (IGF1R)。这些基因的敲除导致尖端细胞的比例以及体外和体内的发芽程度显着降低。总之，这项研究表明，通过使用我们的体外尖端细胞模型，鉴定出了两个新的必需尖端细胞基因。本文的在线版本 (10.1007/s10456-018-9627-4) 包含补充材料，可供授权用户使用。
Tip cells, the leading cells of angiogenic sprouts, were identified in cultures of human umbilical vein endothelial cells (HUVECs) by using CD34 as a marker. Here, we show that tip cells are also present in primary human microvascular endothelial cells (hMVECs), a more relevant endothelial cell type for angiogenesis. By means of flow cytometry, immunocytochemistry, and qPCR, it is shown that endothelial cell cultures contain a dynamic population of CD34+ cells with many hallmarks of tip cells, including filopodia-like extensions, elevated mRNA levels of known tip cell genes, and responsiveness to stimulation with VEGF and inhibition by DLL4. Furthermore, we demonstrate that our in vitro tip cell model can be exploited to investigate cellular and molecular mechanisms in tip cells and to discover novel targets for anti-angiogenesis therapy in patients. Small interfering RNA (siRNA) was used to knockdown gene expression of the known tip cell genes angiopoietin 2 (ANGPT2) and tyrosine kinase with immunoglobulin-like and EGF-like domains 1 (TIE1), which resulted in similar effects on tip cells and sprouting as compared to inhibition of tip cells in vivo. Finally, we identified two novel tip cell-specific genes in CD34+ tip cells in vitro: insulin-like growth factor 2 (IGF2) and IGF-1-receptor (IGF1R). Knockdown of these genes resulted in a significant decrease in the fraction of tip cells and in the extent of sprouting in vitro and in vivo. In conclusion, this study shows that by using our in vitro tip cell model, two novel essential tip cells genes are identified. The online version of this article (10.1007/s10456-018-9627-4) contains supplementary material, which is available to authorized users.
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