C-CBL is required for inhibition of angiogenesis through modulating JAK2/STAT3 activity in ROP development

• Published in: Biomedicine & pharmacotherapy 2020-12, Vol.132, p.110856-110856, Article 110856
• Main Authors: Chen, Shimei, Sun, Qiao, Sun, Dandan, Willette-Brown, Jami, Anderson, Matthew J., Gu, Qing, Lewandoski, Mark, Hu, Yinling, Zhu, Feng, Wei, Fang, Zhang, Jian
• Format: Article
• Language: English
• Subjects: Angiogenesis; Animals; c-Cbl; Cells, Cultured; Disease Models, Animal; Endothelial Cells - pathology; Gene Knockdown Techniques; JAK2/STAT3/VEGF signaling pathway; Janus Kinase 2 - metabolism; Mice; Mice, Inbred C57BL; Oxygen; Proto-Oncogene Proteins c-cbl - genetics; Retinal Neovascularization - genetics; Retinal Neovascularization - pathology; Retinal Vessels - cytology; Retinal Vessels - pathology; Retinopathy of prematurity; Retinopathy of Prematurity - genetics; Retinopathy of Prematurity - pathology; Signal Transduction; STAT3 Transcription Factor - metabolism; Vascular Endothelial Growth Factor A - metabolism
• ISSN: 0753-3322; 1950-6007
• DOI: 10.1016/j.biopha.2020.110856

[Display omitted] •Hypoxia exposure reduces C-CBL protein level in both mouse cells and retinas.•C-CBL controls JAK2/STAT3/VEGF-mediated angiogenesis.•JAK2 inhibitor can reverse neovascularization in mRMECs with c-Cbl knockdown.•Overexpression of c-Cbl blocks neovascularization in the retinas of OIR mice. The incidence of retinopathy of prematurity (ROP) has increased continuously in recent years. However, the therapeutic effects of current treatments still remain undesired. This study aims to investigate the role of C-CBL in retinal angiogenesis in ROP and its potential as a therapeutic target. Mouse retina microvascular endothelial cells (mRMECs) and induced experimental ROP/ oxygen-induced retinopathy (OIR) mice were employed to investigate the role of C-CBL in angiogenesis with combined molecular and cellular approaches, and histopathology methods. OIR mouse pups at postnatal day 12 (P12) were either injected intravitreally with adenovirus overexpressing c-Cbl or c-Cbl siRNA. Retinal neovascularization and avascular status were evaluated by retinal immunofluorescence (IF) staining, whole-mounts and hematoxylin and eosin (H&E) staining. C-CBL inhibits neovascularization by negatively regulating JAK2/STAT3/VEGF signaling axis in a ubiquitination-dependent manner. Knockdown of c-Cbl by siRNA reduced ubiquitin-mediated JAK2 degradation and increased levels of p-JAK2, p-STAT3, VEGF, and neovascularization in mRMECs, which can be reversed by JAK2 inhibitor treatment. While knockdown of c-Cbl significantly increased neovascular (NV) zone in the retinas, c-Cbl overexpression inhibited neovascularization in the retinal tissues in OIR mice. We found that C-CBL is required for anti-neovascularization process in ROP development by inhibiting JAK2/STAT3-dependent angiogenesis. Thus, our finding strongly suggest that C-CBL may be a potential novel therapeutic target for treating ROP.