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Suppression of choroidal neovascularization through inhibition of APE1/Ref-1 redox activity
Yue Li1, Xiuli Liu2, Tongrong Zhou2
1Department of Ophthalmology, Henry Ford Health System, Detroit, Michigan, United States Department of Ophthalmology, Shaanxi Maternity and Child Healthcare Hospital, Xi'an, Shaanxi, People's Republic of China.
Purpose:
The redox function of APE1/Ref-1 is a key regulator in pathological angiogenesis, such as retinal neovascularization and tumor growth. In this study, we examined whether inhibition of APE1/Ref-1 redox function by a small molecule inhibitor E3330 suppresses experimental choroidal neovascularization (CNV) in vitro and in vivo.
Methods:
Primate choroid endothelial cells (CECs) received treatment of 0 to 100 μM E3330 alone or cotreatment of E3330 and 500 μg/mL anti-VEGF antibody bevacizumab. Choroid endothelial cell angiogenic function was examined by cell proliferation, migration, and tube formation assays. The effects of E3330 on NF-κB and STAT3 signaling pathways were determined by reporter gene assay, Western blot, and ELISA. Laser-induced CNV mouse model was used to test the effects of E3330 in vivo. Potential toxicity of E3330 was evaluated by TUNEL assay.
Results:
The E3330 of 25 to 100 μM dose-dependently suppressed CEC proliferation, migration, and tube formation, in the absence of noticeable cell toxicity. Lower doses of E3330 (10-20 μM) reduced the transcriptional activity of NF-κB and STAT3 without affecting protein phosphorylation of both molecules. At the same time, E3330 downregulated MCP-1 production in CECs. The antiangiogenic effect of E3330 was comparable and additive to bevacizumab. The E3330 effectively attenuated the progression of laser-induced CNV in mice after a single intravitreal injection.
Conclusions:
The APE1/Ref-1 redox function regulates multiple transcription factors and inflammatory molecules, and is essential for CEC angiogenesis. Specific inhibition of APE1/Ref-1 redox function with E3330 may represent a promising novel treatment for wet AMD.
Insights
The small molecule E3330 inhibits APE1/Ref-1 redox function, suppressing pathological angiogenesis in choroidal neovascularization (CNV) models. This novel therapeutic approach shows promise for treating wet age-related macular degeneration (AMD).
Area of Science:
- Molecular biology
- Ophthalmology
- Angiogenesis research
Background:
- The redox function of AP endonuclease 1/redox factor 1 (APE1/Ref-1) is crucial in pathological angiogenesis, including retinal neovascularization and tumor growth.
- APE1/Ref-1 regulates key transcription factors and inflammatory molecules involved in cellular processes like proliferation and migration.
Purpose of the Study:
- To investigate the efficacy of E3330, a small molecule inhibitor of APE1/Ref-1 redox function, in suppressing experimental choroidal neovascularization (CNV).
- To evaluate the in vitro and in vivo effects of E3330 on choroid endothelial cell (CEC) angiogenic functions and signaling pathways.
Main Methods:
- Primate choroid endothelial cells (CECs) were treated with E3330 and/or bevacizumab to assess proliferation, migration, and tube formation.
- Reporter gene assays, Western blots, and ELISA were used to determine the effects of E3330 on NF-κB and STAT3 signaling.
- A laser-induced CNV mouse model was employed for in vivo efficacy testing, with toxicity evaluated by TUNEL assay.
Main Results:
- E3330 dose-dependently suppressed CEC proliferation, migration, and tube formation without significant toxicity.
- Low-dose E3330 reduced NF-κB and STAT3 transcriptional activity and downregulated MCP-1 production.
- E3330 demonstrated comparable and additive antiangiogenic effects to bevacizumab and attenuated CNV progression in vivo.
Conclusions:
- APE1/Ref-1 redox function is essential for CEC angiogenesis, regulating multiple transcription factors and inflammatory molecules.
- Specific inhibition of APE1/Ref-1 redox function using E3330 presents a potential novel therapeutic strategy for wet age-related macular degeneration (AMD).
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