Ref-1 redox activity modulates canonical Wnt signaling in endothelial cells
Ischemic retinopathies, including proliferative diabetic retinopathy (PDR) and retinopathy of prematurity (ROP), are characterized by abnormal retinal neovascularization and can lead to blindness in children and adults. Current treatments, such as intravitreal anti-VEGF injections, face limitations...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-06-01
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| Series: | Redox Biology |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2213231725001594 |
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| author | Gabriella D. Hartman Kamakshi Sishtla Eyram K. Kpenu Mahmut Mijit Anbukkarasi Muniyandi Ha-Neul Jo Harald J. Junge Aaron Shaw Daniela Bischof Sheng Liu Jun Wan Mark R. Kelley Timothy W. Corson |
| author_facet | Gabriella D. Hartman Kamakshi Sishtla Eyram K. Kpenu Mahmut Mijit Anbukkarasi Muniyandi Ha-Neul Jo Harald J. Junge Aaron Shaw Daniela Bischof Sheng Liu Jun Wan Mark R. Kelley Timothy W. Corson |
| author_sort | Gabriella D. Hartman |
| collection | DOAJ |
| description | Ischemic retinopathies, including proliferative diabetic retinopathy (PDR) and retinopathy of prematurity (ROP), are characterized by abnormal retinal neovascularization and can lead to blindness in children and adults. Current treatments, such as intravitreal anti-VEGF injections, face limitations due to high treatment burden and variable efficacy, as multiple signaling pathways, beyond VEGF, contribute to retinal neovascularization. Previous studies demonstrate that targeting the redox-mediated transcriptional regulatory function of APE1/Ref-1 reduces pathological neovascularization. We aimed to identify novel signaling pathways regulated by Ref-1 redox activity utilizing RNA sequencing of human retinal endothelial cells (HRECs) treated with a Ref-1 redox inhibitor. We found that Wnt/β-catenin signaling was significantly downregulated after Ref-1 inhibition. Given the role of Wnt signaling in vascular pathologies, we investigated how Ref-1 regulates Wnt/β-catenin signaling. Ref-1 inhibition downregulated Wnt co-receptors LRP5/6 at both the mRNA and protein levels in endothelial cells, suggesting transcriptional regulation. Ref-1 redox inhibitors APX3330 and APX2009 reduced Wnt3a-induced nuclear β-catenin levels, decreased Wnt transcriptional activity by TOPFlash luciferase assay, and blocked hypoxia-induced Wnt/β-catenin activation in HRECs. In the oxygen-induced retinopathy mouse model of retinal neovascularization, Ref-1 specific inhibitor APX2009 reduced the expression of Wnt-related genes at sites of neovascularization. These findings reveal a novel role for Ref-1 redox activity in modulating Wnt/β-catenin signaling in endothelial cells and highlight the potential of Ref-1 redox activity targeted inhibitors as a novel therapeutic approach for retinal neovascular diseases by modulating multiple disease-relevant pathways. |
| format | Article |
| id | doaj-art-d1ae4fa0a5eb4b9cb0cae5bac50de27b |
| institution | OA Journals |
| issn | 2213-2317 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Redox Biology |
| spelling | doaj-art-d1ae4fa0a5eb4b9cb0cae5bac50de27b2025-08-20T02:31:54ZengElsevierRedox Biology2213-23172025-06-018310364610.1016/j.redox.2025.103646Ref-1 redox activity modulates canonical Wnt signaling in endothelial cellsGabriella D. Hartman0Kamakshi Sishtla1Eyram K. Kpenu2Mahmut Mijit3Anbukkarasi Muniyandi4Ha-Neul Jo5Harald J. Junge6Aaron Shaw7Daniela Bischof8Sheng Liu9Jun Wan10Mark R. Kelley11Timothy W. Corson12Department of Ophthalmology, Indiana University School of Medicine, Indianapolis, IN, USA; Stark Neurosciences Research Institute, Indiana University School of Medicine, Indianapolis, IN, USADepartment of Ophthalmology, Indiana University School of Medicine, Indianapolis, IN, USA; Department of Pharmacology and Toxicology, Indiana University School of Medicine, Indianapolis, IN, USA; Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, ON, CanadaHerman B Wells Center for Pediatric Research, Department of Pediatrics, Indiana University School of Medicine, Indianapolis, IN, USA; Indiana University Simon Comprehensive Cancer Center, Indianapolis, IN, USA; Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, IN, USAHerman B Wells Center for Pediatric Research, Department of Pediatrics, Indiana University School of Medicine, Indianapolis, IN, USA; Indiana University Simon Comprehensive Cancer Center, Indianapolis, IN, USADepartment of Ophthalmology, Indiana University School of Medicine, Indianapolis, IN, USA; Department of Pharmacology and Toxicology, Indiana University School of Medicine, Indianapolis, IN, USADepartment of Ophthalmology and Visual Neuroscience, University of Minnesota, Minneapolis, MN, USADepartment of Ophthalmology and Visual Neuroscience, University of Minnesota, Minneapolis, MN, USADepartment of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, IN, USADepartment of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, IN, USACenter for Computational Biology and Bioinformatics, Indiana University, Indianapolis, IN, USADepartment of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, IN, USA; Center for Computational Biology and Bioinformatics, Indiana University, Indianapolis, IN, USADepartment of Ophthalmology, Indiana University School of Medicine, Indianapolis, IN, USA; Department of Pharmacology and Toxicology, Indiana University School of Medicine, Indianapolis, IN, USA; Herman B Wells Center for Pediatric Research, Department of Pediatrics, Indiana University School of Medicine, Indianapolis, IN, USA; Indiana University Simon Comprehensive Cancer Center, Indianapolis, IN, USA; Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, IN, USA; Corresponding author. Department of Pediatrics, Walther Hall R3, C218F 980 W. Walnut Street, Indianapolis, IN, 46202, USA.Department of Ophthalmology, Indiana University School of Medicine, Indianapolis, IN, USA; Stark Neurosciences Research Institute, Indiana University School of Medicine, Indianapolis, IN, USA; Department of Pharmacology and Toxicology, Indiana University School of Medicine, Indianapolis, IN, USA; Indiana University Simon Comprehensive Cancer Center, Indianapolis, IN, USA; Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, IN, USA; Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, ON, Canada; Department of Ophthalmology and Vision Sciences, Temerty Faculty of Medicine, University of Toronto, Toronto, ON, Canada; Corresponding author. Leslie Dan Faculty of Pharmacy, University of Toronto, 144 College Street, Toronto, Ontario, M5S 3M2, Canada.Ischemic retinopathies, including proliferative diabetic retinopathy (PDR) and retinopathy of prematurity (ROP), are characterized by abnormal retinal neovascularization and can lead to blindness in children and adults. Current treatments, such as intravitreal anti-VEGF injections, face limitations due to high treatment burden and variable efficacy, as multiple signaling pathways, beyond VEGF, contribute to retinal neovascularization. Previous studies demonstrate that targeting the redox-mediated transcriptional regulatory function of APE1/Ref-1 reduces pathological neovascularization. We aimed to identify novel signaling pathways regulated by Ref-1 redox activity utilizing RNA sequencing of human retinal endothelial cells (HRECs) treated with a Ref-1 redox inhibitor. We found that Wnt/β-catenin signaling was significantly downregulated after Ref-1 inhibition. Given the role of Wnt signaling in vascular pathologies, we investigated how Ref-1 regulates Wnt/β-catenin signaling. Ref-1 inhibition downregulated Wnt co-receptors LRP5/6 at both the mRNA and protein levels in endothelial cells, suggesting transcriptional regulation. Ref-1 redox inhibitors APX3330 and APX2009 reduced Wnt3a-induced nuclear β-catenin levels, decreased Wnt transcriptional activity by TOPFlash luciferase assay, and blocked hypoxia-induced Wnt/β-catenin activation in HRECs. In the oxygen-induced retinopathy mouse model of retinal neovascularization, Ref-1 specific inhibitor APX2009 reduced the expression of Wnt-related genes at sites of neovascularization. These findings reveal a novel role for Ref-1 redox activity in modulating Wnt/β-catenin signaling in endothelial cells and highlight the potential of Ref-1 redox activity targeted inhibitors as a novel therapeutic approach for retinal neovascular diseases by modulating multiple disease-relevant pathways.http://www.sciencedirect.com/science/article/pii/S2213231725001594Wnt signalingAPE1/Ref-1Retinal neovascularizationIschemic retinopathyLRP5Ref-1 redox activity |
| spellingShingle | Gabriella D. Hartman Kamakshi Sishtla Eyram K. Kpenu Mahmut Mijit Anbukkarasi Muniyandi Ha-Neul Jo Harald J. Junge Aaron Shaw Daniela Bischof Sheng Liu Jun Wan Mark R. Kelley Timothy W. Corson Ref-1 redox activity modulates canonical Wnt signaling in endothelial cells Redox Biology Wnt signaling APE1/Ref-1 Retinal neovascularization Ischemic retinopathy LRP5 Ref-1 redox activity |
| title | Ref-1 redox activity modulates canonical Wnt signaling in endothelial cells |
| title_full | Ref-1 redox activity modulates canonical Wnt signaling in endothelial cells |
| title_fullStr | Ref-1 redox activity modulates canonical Wnt signaling in endothelial cells |
| title_full_unstemmed | Ref-1 redox activity modulates canonical Wnt signaling in endothelial cells |
| title_short | Ref-1 redox activity modulates canonical Wnt signaling in endothelial cells |
| title_sort | ref 1 redox activity modulates canonical wnt signaling in endothelial cells |
| topic | Wnt signaling APE1/Ref-1 Retinal neovascularization Ischemic retinopathy LRP5 Ref-1 redox activity |
| url | http://www.sciencedirect.com/science/article/pii/S2213231725001594 |
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