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

Shimei Chen; Qiao Sun; Dandan Sun; Jami Willette-Brown; Matthew J Anderson; Qing Gu; Mark Lewandoski; Yinling Hu; Feng Zhu; Fang Wei; Jian Zhang

doi:10.1016/j.biopha.2020.110856

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

Biomed Pharmacother. 2020 Dec:132:110856. doi: 10.1016/j.biopha.2020.110856. Epub 2020 Oct 28.

Authors

Shimei Chen¹, Qiao Sun¹, Dandan Sun¹, Jami Willette-Brown², Matthew J Anderson³, Qing Gu⁴, Mark Lewandoski³, Yinling Hu², Feng Zhu⁵, Fang Wei⁶, Jian Zhang⁷

Affiliations

¹ Department of Ophthalmology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China; Shanghai Key Laboratory of Ocular Fundus Diseases, Shanghai, 200080, China; Shanghai Engineering Center for Visual Science and Photomedicine, Shanghai, 200080, China; National Clinical Research Center for Eye Diseases, Shanghai, 20080, China; Shanghai Engineering Center for Precise Diagnosis and Treatment of Eye Diseases, Shanghai, 20080, China.
² Laboratory of Cancer Immunometabolism, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, MD, 21702, USA.
³ Cancer and Developmental Biology Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, MD, 21702, USA.
⁴ Department of Ophthalmology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China; Shanghai Key Laboratory of Ocular Fundus Diseases, Shanghai, 200080, China; Shanghai Engineering Center for Visual Science and Photomedicine, Shanghai, 200080, China.
⁵ Laboratory of Cancer Immunometabolism, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, MD, 21702, USA. Electronic address: [email protected].
⁶ Department of Ophthalmology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China; Shanghai Key Laboratory of Ocular Fundus Diseases, Shanghai, 200080, China; Shanghai Engineering Center for Visual Science and Photomedicine, Shanghai, 200080, China; National Clinical Research Center for Eye Diseases, Shanghai, 20080, China; Shanghai Engineering Center for Precise Diagnosis and Treatment of Eye Diseases, Shanghai, 20080, China. Electronic address: [email protected].
⁷ Department of Ophthalmology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China; Shanghai Key Laboratory of Ocular Fundus Diseases, Shanghai, 200080, China; Shanghai Engineering Center for Visual Science and Photomedicine, Shanghai, 200080, China; National Clinical Research Center for Eye Diseases, Shanghai, 20080, China; Shanghai Engineering Center for Precise Diagnosis and Treatment of Eye Diseases, Shanghai, 20080, China. Electronic address: [email protected].

Abstract

Purpose: 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.

Methods: 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.

Results: 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.

Conclusion: 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.

Keywords: Angiogenesis; JAK2/STAT3/VEGF signaling pathway; Retinopathy of prematurity; c-Cbl.

MeSH terms

Animals
Cells, Cultured
Disease Models, Animal
Endothelial Cells / pathology
Gene Knockdown Techniques
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 / genetics
Retinopathy of Prematurity / pathology*
STAT3 Transcription Factor / metabolism
Signal Transduction
Vascular Endothelial Growth Factor A / metabolism

Substances

STAT3 Transcription Factor
Stat3 protein, mouse
Vascular Endothelial Growth Factor A
vascular endothelial growth factor A, mouse
Proto-Oncogene Proteins c-cbl
Jak2 protein, mouse
Janus Kinase 2
Cbl protein, mouse
Oxygen

Abstract

MeSH terms

Substances

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