title: Suppression of choroidal neovascularization through inhibition of APE1/Ref-1 redox activity.
abstract: OBJECTIVE
  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.
authors: Department 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.
authors: Yue
authors: Y
authors: Li
authors: Department of Ophthalmology, Henry Ford Health System, Detroit, Michigan, United States.
authors: Xiuli
authors: X
authors: Liu
authors: Department of Ophthalmology, Henry Ford Health System, Detroit, Michigan, United States.
authors: Tongrong
authors: T
authors: Zhou
authors: Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, Indiana, United States.
authors: Mark R
authors: MR
authors: Kelley
authors: Department of Ophthalmology, Henry Ford Health System, Detroit, Michigan, United States.
authors: Paul A
authors: PA
authors: Edwards
authors: Department of Ophthalmology, Henry Ford Health System, Detroit, Michigan, United States.
authors: Hua
authors: H
authors: Gao
authors: Department of Ophthalmology, Henry Ford Health System, Detroit, Michigan, United States.
authors: Xiaoxi
authors: X
authors: Qiao
mesh_terms: D000818:Animals; D061067:Antibodies, Monoclonal, Humanized / Q000494:pharmacology; D016227:Benzoquinones / Q000494:pharmacology*; D000068258:Bevacizumab; D015153:Blotting, Western; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D018932:Chemokine CCL2 / Q000378:metabolism; D002829:Choroid / Q000098:blood supply; D020256:Choroidal Neovascularization / Q000473:pathology / Q000517:prevention & control*; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase / Q000037:antagonists & inhibitors*; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D020287:In Situ Nick-End Labeling; D058449:Intravitreal Injections; D008253:Macaca mulatta; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D016328:NF-kappa B / Q000378:metabolism; D011422:Propionates / Q000494:pharmacology*; D050796:STAT3 Transcription Factor / Q000378:metabolism
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D016227:Benzoquinones; D018932:Chemokine CCL2; D016328:NF-kappa B; D011422:Propionates; D050796:STAT3 Transcription Factor; C494087:Stat3 protein, mouse; C075569:E 3330; D000068258:Bevacizumab; C486216:Apex1 protein, mouse; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase
keywords: APE1/Ref-1; antiangiogenesis; choroidal neovascularization; redox function; transcription factors