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Exploring the Efficacy of AZD6738 in Corneal Neovascularization: Autophagy Enhancement and Angiogenesis Inhibition
Qing Wang1, Yangyang Peng1, Shuna Wang2
1The Second Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, China.
Abstract:
Purpose: This study aimed to investigate the therapeutic potential of AZD6738, an ataxia-telangiectasia and rad3-related (ATR) kinase inhibitor, in preventing corneal neovascularization (CNV) by exploring its effects on autophagy regulation and angiogenesis. Methods: Human umbilical vein endothelial cells were cultured and treated with varying concentrations of AZD6738 and vascular endothelial growth factor (VEGF) to assess cell viability, migration, and tube formation. A corneal alkali burn model in Sprague-Dawley rats was established to evaluate the in vivo effects of AZD6738 on CNV. Autophagy was assessed using monodansylcadaverine (MDC) staining, western blotting, and qRT-PCR to measure the expression of autophagy-related markers and key proteins involved in the PI3K-AKT pathway. Immunohistochemistry and immunofluorescence staining were employed to examine histological changes and the expression of markers related to neovascularization and fibrosis. Results: The study demonstrated that AZD6738 significantly inhibited cell viability in a dose-dependent manner. AZD6738 effectively reduced VEGF-induced cell migration and tube formation. Moreover, the introduction of AZD6738 enhanced autophagy, as indicated by increased MDC staining, upregulated Beclin1 expression, and an elevated LC3 II/I ratio. The inhibitor also suppressed the PI3K-AKT pathway, reducing VEGF and VEGFR2 expression, and decreasing the phosphorylation levels of AMPK and AKT. In an experimental CNV model, AZD6738 treatment resulted in a significant reduction in CNV, with fewer and shorter blood vessels observed, as well as changes in autophagy-related proteins. Conclusions: AZD6738 showed potential in preventing CNV. Its ability to enhance autophagy and inhibit PI3K-AKT-VEGF pathways in angiogenesis suggests that AZD6738 could be an effective treatment strategy for CNV.
Insights
AZD6738, an ATR kinase inhibitor, effectively prevents corneal neovascularization (CNV) by enhancing autophagy and inhibiting angiogenesis pathways. This study highlights its therapeutic potential for CNV treatment.
Area of Science:
- Ophthalmology
- Molecular Biology
- Pharmacology
Background:
- Corneal neovascularization (CNV) is a pathological process involving new blood vessel growth in the cornea.
- Current treatments for CNV have limitations, necessitating the exploration of novel therapeutic strategies.
- Understanding the molecular mechanisms underlying CNV, including autophagy and angiogenesis, is crucial for developing effective interventions.
Purpose of the Study:
- To investigate the therapeutic potential of AZD6738, an ataxia-telangiectasia and rad3-related (ATR) kinase inhibitor, in preventing corneal neovascularization (CNV).
- To explore the effects of AZD6738 on autophagy regulation and angiogenesis in the context of CNV.
- To evaluate the impact of AZD6738 on key signaling pathways involved in neovascularization.
Main Methods:
- * In vitro: Assessed AZD6738 effects on human umbilical vein endothelial cell viability, migration, and tube formation, with or without vascular endothelial growth factor (VEGF).
- * In vivo: Utilized a rat corneal alkali burn model to evaluate AZD6738's efficacy in preventing CNV.
- * Molecular analysis: Measured autophagy markers (MDC staining, Beclin1, LC3 II/I ratio) and PI3K-AKT pathway proteins (VEGF, VEGFR2, p-AMPK, p-AKT) via western blotting and qRT-PCR.
- * Histological assessment: Employed immunohistochemistry and immunofluorescence to examine CNV and fibrosis markers.
Main Results:
- AZD6738 significantly inhibited endothelial cell viability, migration, and VEGF-induced tube formation in vitro.
- The ATR inhibitor enhanced autophagy, evidenced by increased MDC staining and upregulated Beclin1 and LC3 II/I ratio.
- AZD6738 suppressed the PI3K-AKT pathway, reducing VEGF and VEGFR2 expression, and decreasing AMPK and AKT phosphorylation, leading to reduced CNV in vivo.
Conclusions:
- AZD6738 demonstrates significant potential as a therapeutic agent for preventing corneal neovascularization.
- The mechanism involves enhancing autophagy and inhibiting the PI3K-AKT-VEGF signaling pathway crucial for angiogenesis.
- AZD6738 represents a promising novel treatment strategy for CNV, warranting further clinical investigation.
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