Inhibition of EZH2 alleviates angiogenesis in a model of corneal neovascularization by blocking FoxO3a‐mediated oxidative stress

• Published in: The FASEB journal 2020-08, Vol.34 (8), p.10168-10181
• Main Authors: Wan, Shan‐Shan, Pan, Yu‐Miao, Yang, Wan‐Ju, Rao, Zhuo‐Qun, Yang, Yan‐Ning
• Format: Article
• Language: English
• Subjects: Adenosine - analogs & derivatives; Adenosine - pharmacology; angiogenesis; Animals; Cells, Cultured; cornea; Corneal Neovascularization - drug therapy; Corneal Neovascularization - metabolism; Disease Models, Animal; enhancer of zeste homolog 2; Enhancer of Zeste Homolog 2 Protein - antagonists & inhibitors; Enhancer of Zeste Homolog 2 Protein - metabolism; Forkhead Box Protein O3 - metabolism; Human Umbilical Vein Endothelial Cells; Humans; Hypoxia - metabolism; Male; Mice; Mice, Inbred BALB C; Neovascularization, Pathologic - drug therapy; Neovascularization, Pathologic - metabolism; oxidative stress; Oxidative Stress - drug effects; Oxidative Stress - physiology; Phosphatidylinositol 3-Kinases - metabolism; Proto-Oncogene Proteins c-akt - metabolism; Reactive Oxygen Species - metabolism; RNA, Small Interfering - metabolism; Signal Transduction - drug effects; Signal Transduction - physiology; Transcription, Genetic - drug effects; Transcription, Genetic - physiology
• ISSN: 0892-6638; 1530-6860
• DOI: 10.1096/fj.201902814RRR

Enhancer of zeste homolog 2 (EZH2), a well‐known methyltransferase, mediates histone H3 lysine 27 trimethylation (H3K27me3) and plays a vital role in ophthalmological disease. However, its role in corneal neovascularization (CoNV) remains unclear. In vitro and in vivo models were assessed in hypoxia‐stimulated angiogenesis and in a mouse model of alkali burn‐induced CoNV. Human umbilical vein endothelial cells (HUVECs) were cultured under hypoxic conditions and different reoxygenation times to identify the molecular mechanisms involved in this process. In this study, we found that EZH2 was positively related to corneal alkali burn‐induced injury. Inhibition of EZH2 with 3‐Deazaneplanocin A (DZNeP) alleviated corneal injury, including oxidative stress and neovascularization in vivo. Similarly, inhibition of EZH2 with either DZNeP or small interfering RNA (siRNA) exerted an inhibitory effect on hypoxia/reoxygenation (H/R)‐induced oxidative stress and angiogenesis in HUVECs. Moreover, our study revealed that ablation of reactive oxygen species (ROS) with N‐acetyl‐cysteine suppressed angiogenesis in HUVECs exposed to H/R stimulation. Furthermore, Forkhead‐box protein O3a (FoxO3a), which was positively associated with ROS production and angiogenesis, was elevated during H/R. This effect could be reversed through the suppression of the transcription activity of EZH2 with DZNeP or siRNA. In addition, the PI3K/Akt pathway, which is the upstream of FoxO3a, was activated in both DZNeP‐treated mice and EZH2‐inhibited HUVECs. Collectively, our results demonstrated that the inhibition of EZH2 alleviated corneal angiogenesis by inhibiting FoxO3a‐dependent ROS production through the PI3K/Akt signaling pathway. These findings indicate that EZH2 may be a valuable therapeutic target for CoNV.