Ref-1 in Retinal Neovascularization

Ref-1 在视网膜新生血管中的作用

• 批准号: 10679621
• 负责人: Gabriella Hartman
• 金额: $ 3.6万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-14 至 2026-06-13
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10679621
• 关键词: Adult; Age; Angiogenesis Inhibitors; Area; Biological Assay; Biological Factors; Blindness; Child; Choroidal Neovascularization; Co-Immunoprecipitations; Complex; Cysteine; Data; Deoxyribonuclease I; Diabetes Mellitus; Diabetic Retinopathy; Disease; Down-Regulation; Endothelial Cells; Exhibits; Eye; Eye diseases; FZD1 gene; FZD4 gene; Gene Expression; Genes; Genetic; Goals; HIF1A gene; Human; Immunofluorescence Immunologic; Inflammatory; Insulin-Dependent Diabetes Mellitus; Knowledge; Lasers; Lead; Link; Medical; Modeling; Molecular; Mus; Oral; Oral Administration; Oxidation-Reduction; Oxygen; Pathogenesis; Pathologic Neovascularization; Pathology; Pathway interactions; Patients; Pharmacological Treatment; Prevalence; Proteins; Quality of life; Regulation; Reporter; Research; Resistance; Retina; Retinal Diseases; Retinal Neovascularization; Retinopathy of Prematurity; Role; STAT3 gene; Severity of illness; Signal Transduction; Societies; Testing; Therapeutic; Therapeutic Effect; Tissues; Treatment Factor; Treatment Protocols; Up-Regulation; Vascular Diseases; Vascular Endothelial Growth Factors; Vision; Visual impairment; WNT Signaling Pathway; Western Blotting; angiogenesis; combat; endonuclease; human RNA sequencing; improved; in vitro Assay; in vivo; inhibitor; insight; interest; intravitreal injection; mRNA Expression; mouse model; neovascular; neovascularization; new therapeutic target; novel; novel therapeutic intervention; novel therapeutics; oxidation; pharmacologic; preterm newborn; proliferative diabetic retinopathy; protein expression; receptor; small molecule; small molecule inhibitor; spatiotemporal; therapy resistant; transcription factor; transcriptome sequencing; treatment response

PROJECT SUMMARY Retinal neovascularization impairs visual function and is a hallmark of several neovascular eye diseases, including retinopathy of prematurity (ROP) and proliferative diabetic retinopathy (PDR). Diabetic retinopathy (DR) is the leading cause of vision-loss in working age adults, and ROP is the leading cause of preventable blindness in children. Current treatments include intravitreal (IVT) injections of anti-vascular endothelial growth factor (VEGF) biologics. However, these therapeutics are often accompanied by high treatment burden and resistance to therapy. Previous research indicates that VEGF alone is not sufficient for induction of neovascularization, suggesting that multiple disease-relevant pathways may be targeted to increase therapeutic response. Thus, there is a critical need to develop novel therapies that modulate multiple disease- relevant pathways and circumvent the issues with IVT injections. Prior studies indicate that APE1/Ref-1, a multifunctional protein with both endonuclease (APE1) and redox activity (Ref-1), regulates multiple transcription factors that are linked to retinal neovascularization. Preliminary data in the laser-induced choroidal neovascularization model demonstrated upregulation of Ref-1 during neovascularization, and oral administration of a small molecule Ref-1 redox inhibitor decreased disease severity. RNA-seq of human retinal endothelial cells revealed that a Ref-1 redox inhibitor downregulated Wnt signaling genes, implying that Ref-1 redox function modulates neovascularization through the Wnt signaling. But, the underlying molecular mechanisms of the novel Ref-1 target in retinal neovascularization have yet to be elucidated. Identifying the molecular mechanism underlying Ref-1 in retinal neovascularization is critical in developing novel therapies to reduce vision loss. The hypothesis of this proposal is that Ref-1 redox activity promotes induction of retinal neovascularization via activation of the Wnt signaling pathway, and reducing Ref-1 redox activity with a novel Ref-1 redox inhibitor will exhibit potent therapeutic effects via inhibition of the Wnt signaling pathway. Aim 1 will analyze the spatiotemporal expression of Ref-1 in retinal neovascularization in the oxygen-induced retinopathy (OIR) mouse model, the Vldlr-/- mouse model, and in human PDR tissue. Aim 2 will analyze the mechanism of Ref-1 regulation of the Wnt pathway in retinal endothelial cells through functional in vitro assays. Aim 3 will assess anti-angiogenic effects of a Ref-1 inhibitor in vivo by administering oral Ref-1 inhibitor in the OIR mouse model and evaluating changes in neovascularization. Effects on the Wnt signaling pathway will also be assessed using immunofluorescence. Completion of this study will bolster understanding of Ref-1 in neovascular eye diseases and provide a novel therapeutic strategy to combat vision loss and blindness due to these diseases.

项目摘要 视网膜新生血管形成损害视功能，是几种新生血管性眼病的标志， 包括早产儿视网膜病变（ROP）和增殖性糖尿病视网膜病变（PDR）。糖尿病视网膜病变 (DR)是工作年龄成年人视力丧失的主要原因，而ROP是可预防的主要原因。 儿童失明。目前的治疗方法包括玻璃体内注射抗血管内皮生长 因子（VEGF）生物制剂。然而，这些疗法通常伴随着高治疗负担， 对治疗的抵抗先前的研究表明，单独的VEGF不足以诱导 新血管形成，这表明多种疾病相关途径可能是有针对性的，以增加 治疗反应。因此，迫切需要开发调节多种疾病的新疗法- 相关途径并规避IVT注射的问题。先前的研究表明，APE 1/Ref-1，a 具有核酸内切酶（APE 1）和氧化还原活性（Ref-1）的多功能蛋白，调节多种 与视网膜新生血管形成有关的转录因子。激光诱导脉络膜 新血管形成模型显示在新血管形成期间Ref-1上调，并且口服 施用小分子Ref-1氧化还原抑制剂降低了疾病的严重程度。人视网膜的RNA测序 内皮细胞显示Ref-1氧化还原抑制剂下调Wnt信号转导基因，这意味着Ref-1 氧化还原功能通过Wnt信号传导调节新血管形成。但是， 新的Ref-1靶点在视网膜新生血管形成中的作用机制尚未阐明。识别 Ref-1在视网膜新生血管形成中的分子机制对于开发新的治疗方法至关重要， 减少视力丧失。该提议的假设是Ref-1氧化还原活性促进了视网膜色素变性的诱导。 通过激活Wnt信号传导途径，并用新的抗氧化剂降低Ref-1的氧化还原活性， Ref-1氧化还原抑制剂将通过抑制Wnt信号传导途径表现出有效的治疗效果。目标1将 分析氧诱导视网膜病变视网膜新生血管中Ref-1的时空表达 (OIR)小鼠模型、Vldlr-/-小鼠模型和人PDR组织中。目标2将分析 通过功能性体外测定，Ref-1对视网膜内皮细胞中Wnt通路的调节。目标3将 通过在OIR小鼠中给予口服Ref-1抑制剂来评估Ref-1抑制剂的体内抗血管生成作用 模型和评估新血管形成的变化。对Wnt信号通路的影响也将被 使用免疫荧光进行评估。本研究的完成将加强对Ref-1的理解， 新生血管性眼病，并提供了一种新的治疗策略，以打击视力丧失和失明， 这些疾病。