VEGF Induces Expression of Genes That Either Promote or Limit Relaxation of the Retinal Endothelial Barrier

The purpose of this study was to identify genes that mediate VEGF-induced permeability. We performed RNA-Seq analysis on primary human retinal endothelial cells (HRECs) cultured in normal (5 mM) and high glucose (30 mM) conditions that were treated with vehicle, VEGF, or VEGF then anti-VEGF. We filtered our RNA-Seq dataset to identify genes with the following four characteristics: (1) regulated by VEGF, (2) VEGF regulation reversed by anti-VEGF, (3) regulated by VEGF in both normal and high glucose conditions, and (4) known contribution to vascular homeostasis. Of the resultant 18 genes, members of the Notch signaling pathway and ANGPT2 (Ang2) were selected for further study. Permeability assays revealed that while the Notch pathway was dispensable for relaxing the barrier, it contributed to maintaining an open barrier. In contrast, Ang2 limited the extent of barrier relaxation in response to VEGF. These findings indicate that VEGF engages distinct sets of genes to induce and sustain barrier relaxation. Furthermore, VEGF induces expression of genes that limit the extent of barrier relaxation. Together, these observations begin to elucidate the elegance of VEGF-mediated transcriptional regulation of permeability.

Automated summary

The goal of this study was to identify genes involved in VEGF-induced permeability. RNA-Seq analysis was performed on primary human retinal endothelial cells treated with VEGF or VEGF plus anti-VEGF under normal and high glucose conditions. Genes that met specific criteria related to VEGF regulation and vascular homeostasis were identified, including members of the Notch signaling pathway and ANGPT2. The findings indicate that VEGF activates different sets of genes to induce and sustain barrier relaxation, with Notch pathway contributing to maintaining an open barrier and Ang2 limiting the extent of barrier relaxation.