Jo Dong Hyun, Bae Jingi, Chae Sehyun, Kim Jin Hyoung, Han Jong-Hee, Hwang Daehee, Lee Sang-Won, Kim Jeong Hun
From the ‡Fight Against Angiogenesis-related Blindness Laboratory, Clinical Research Institute, Seoul National University Hospital, Seoul 03080, Korea; §Department of Biomedical Sciences and Protein Metabolism Medical Research Center, College of Medicine, Seoul National University, Seoul 03080, Korea;
‖Department of Chemistry, Research Institute for Natural Sciences, Korea University, Seoul 02841, Korea;
Mol Cell Proteomics. 2016 May;15(5):1681-91. doi: 10.1074/mcp.M115.053249. Epub 2016 Mar 11.
Retinal vascular hyperpermeability causes macular edema, leading to visual deterioration in retinal diseases such as diabetic retinopathy and retinal vascular occlusion. Dysregulation of junction integrity between endothelial cells by vascular endothelial growth factor (VEGF) was shown to cause retinal vascular hyperpermeability. Accordingly, anti-VEGF agents have been used to treat retinal vascular hyperpermeability. However, they can confer potential toxicity through their deleterious effects on maintenance and survival of neuronal and endothelial cells in the retina. Thus, it is important to identify novel therapeutic targets for retinal vascular hyperpermeability other than VEGF. Here, we prepared murine retinas showing VEGF-induced vascular leakage from superficial retinal vascular plexus and prevention of VEGF-induced leakage by anti-VEGF antibody treatment. We then performed comprehensive proteome profiling of these samples and identified retinal proteins for which abundances were differentially expressed by VEGF, but such alterations were inhibited by anti-VEGF antibody. Functional enrichment and network analyses of these proteins revealed the β2 integrin pathway, which can prevent dysregulation of junction integrity between endothelial cells through cytoskeletal rearrangement, as a potential therapeutic target for retinal vascular hyperpermeability. Finally, we experimentally demonstrated that inhibition of the β2 integrin pathway salvaged VEGF-induced retinal vascular hyperpermeability, supporting its validity as an alternative therapeutic target to anti-VEGF agents.
视网膜血管高通透性会导致黄斑水肿,进而致使糖尿病视网膜病变和视网膜血管阻塞等视网膜疾病的视力下降。血管内皮生长因子(VEGF)导致内皮细胞间连接完整性失调,进而引发视网膜血管高通透性。因此,抗VEGF药物已被用于治疗视网膜血管高通透性。然而,它们可能通过对视网膜中神经元和内皮细胞的维持与存活产生有害影响而带来潜在毒性。所以,确定除VEGF之外的视网膜血管高通透性新治疗靶点很重要。在此,我们制备了显示VEGF诱导视网膜浅表层血管丛血管渗漏以及抗VEGF抗体治疗可预防VEGF诱导渗漏的小鼠视网膜。然后,我们对这些样本进行了全面的蛋白质组分析,鉴定出VEGF使其丰度差异表达但这种改变被抗VEGF抗体抑制的视网膜蛋白。对这些蛋白质的功能富集和网络分析揭示,β2整合素途径可通过细胞骨架重排防止内皮细胞间连接完整性失调,是视网膜血管高通透性的潜在治疗靶点。最后,我们通过实验证明抑制β2整合素途径可挽救VEGF诱导的视网膜血管高通透性,支持其作为抗VEGF药物替代治疗靶点的有效性。