Department of Pathology and Pathophysiology, Zhejiang University School of Medicine, Hangzhou, China.
Department of Gynecology, Women's Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
FASEB J. 2019 Jan;33(1):1124-1137. doi: 10.1096/fj.201800284R. Epub 2018 Aug 13.
Vascular endothelial (VE)-cadherin junctional localization is known to play a central role in vascular development, endothelial barrier integrity, and homeostasis. The sarcoma homology domain containing protein tyrosine phosphatase (SHP)2 has been shown to be involved in regulating endothelial barrier function; however, the mechanisms remain largely unknown. In this work SHP2 knockdown in an HUVEC monolayer increased VE-cadherin internalization and endothelial barrier permeability. Loss of SHP2 specifically augmented the GTPase activity of ADP-ribosylation factor (ARF)-1. ARF1 knockdown or inhibition of its guanine nucleotide exchange factors (GEFs) markedly attenuated VE-cadherin internalization and barrier hyperpermeability induced by SHP2 deficiency. SHP2 knockdown increased the total and phosphorylated levels of MET, whose activity was necessary for ARF1 activation and VE-cadherin internalization. Furthermore, constitutive endothelium-specific deletion of Shp2 in mice led to disrupted endothelial cell junctions, massive hemorrhage, and lethality in embryos. Induced and endothelium-specific deletion of Shp2 in adult mice resulted in lung hyperpermeability. Inhibitors for ARF1-GEF or MET used in pregnant mice prevented the vascular leakage in endothelial Shp2-deleted embryos. Together, our findings define a novel role of SHP2 in stabilizing junctional VE-cadherin in the resting endothelial barrier through suppressing MET and ARF1 activation.-Zhang, J., Huang, J., Qi, T., Huang, Y., Lu, Y., Zhan, T., Gong, H., Zhu, Z., Shi, Y., Zhou, J., Yu, L., Zhang, X., Cheng, H., Ke, Y. SHP2 protects endothelial cell barrier through suppressing VE-cadherin internalization regulated by MET-ARF1.
血管内皮 (VE)-钙黏蛋白连接定位被认为在血管发育、内皮屏障完整性和稳态中发挥核心作用。已经表明肉瘤同源结构域包含蛋白酪氨酸磷酸酶 (SHP)2 参与调节内皮屏障功能；然而，机制在很大程度上仍然未知。在这项工作中，在 HUVEC 单层中敲低 SHP2 会增加 VE-钙黏蛋白内化和内皮屏障通透性。SHP2 的缺失特异性增强了 ADP-核糖基化因子 (ARF)-1 的 GTPase 活性。ARF1 敲低或抑制其鸟嘌呤核苷酸交换因子 (GEF) 显著减弱了 SHP2 缺乏引起的 VE-钙黏蛋白内化和屏障通透性增加。SHP2 敲低增加了 MET 的总水平和磷酸化水平，其活性对于 ARF1 激活和 VE-钙黏蛋白内化是必要的。此外，在小鼠中内皮细胞特异性敲除 Shp2 导致内皮细胞连接中断、胚胎大量出血和死亡。在成年小鼠中诱导和内皮细胞特异性敲除 Shp2 导致肺通透性增加。在妊娠小鼠中使用 ARF1-GEF 或 MET 的抑制剂可防止内皮 Shp2 缺失胚胎中的血管渗漏。总之，我们的研究结果定义了 SHP2 通过抑制 MET 和 ARF1 激活在静息内皮屏障中稳定连接 VE-钙黏蛋白的新作用。-Zhang, J., Huang, J., Qi, T., Huang, Y., Lu, Y., Zhan, T., Gong, H., Zhu, Z., Shi, Y., Zhou, J., Yu, L., Zhang, X., Cheng, H., Ke, Y. SHP2 通过抑制 MET-ARF1 调节的 VE-钙黏蛋白内化来保护内皮细胞屏障。
