Targeting the Ref-1 signaling node for treating ocular neovascularization

靶向 Ref-1 信号节点治疗眼部新生血管

基本信息

批准号：
10223319
负责人：
Timothy W Corson
金额：
$ 44.54万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10223319
关键词：
Address Age related macular degeneration Angiogenesis Inhibitors Angiogenic Factor Biological Assay Biology Blindness Cells Choroid Choroidal Neovascularization DNA Binding DNA Repair Data Development Disease Drug Kinetics Effectiveness Endothelial Cells Enzymes Eye Eye diseases Generations Genes Genetic Goals Growth Factor Human Hypoxia Hypoxia Inducible Factor In Vitro Inflammation Inflammatory Knowledge Lasers Longevity Mediator of activation protein Mission Modeling Mus National Eye Institute New Agents Oral Outcome Study Oxidation-Reduction Oxygen Pathway interactions Patient Care Patients Pharmaceutical Preparations Phase Play Protein Inhibition Proteins Public Health Quality of life Regulation Research Research Support Retina Retinal Diseases Retinal Neovascularization Role Safety Signal Pathway Signal Transduction Small Interfering RNA Solid Neoplasm Systemic Therapy TNF gene Testing Therapeutic Therapeutic Intervention Time Toxic effect Translations Tube Up-Regulation Vascular Endothelial Growth Factors Visual impairment WNT Signaling Pathway Work angiogenesis bench to bedside chemokine effective therapy hypoxia inducible factor 1 improved in vivo in vivo Model inhibitor/antagonist innovation knock-down migration mutant neovascular neovascularization new therapeutic target novel novel therapeutics ocular angiogenesis ocular neovascularization overexpression pre-clinical prevent proliferative diabetic retinopathy small molecule small molecule inhibitor success synergism targeted agent targeted treatment transcription factor transcriptome sequencing

项目摘要

The neovascular eye diseases proliferative diabetic retinopathy and wet age-related macular degeneration are major causes of blindness through the lifespan. There is thus a critical need to find novel cellular components that could be targeted to block ocular neovascularization. The protein Ref-1 is one such component, responsible for activating redox-dependent transcription factors important for angiogenesis, and overexpressed in neovascularization. Inhibition of Ref-1’s redox function with novel small molecules blocks proliferation of ocular endothelial cells in vitro and in vivo in the laser-induced choroidal neovascularization (L-CNV) model. Ref-1 inhibition reduces signaling through hypoxia and inflammation pathways, and preliminary data reveals a novel connection to Wnt signaling. The long-term goal is to elucidate the role of Ref-1 in ocular neovascularization, and develop novel therapies targeting this protein or its pathway(s). The rationale for this research is that Ref-1 is a significant mediator of angiogenesis and inflammation, a target of multiple antiangiogenic small molecules, and a regulator of key angiogenesis factors including hypoxia-inducible factor 1α and NF-κB. The objectives in this application are to determine how Ref-1 functions as a regulator of angiogenesis and to develop new agents targeting this enzyme. The overall hypothesis is that Ref-1 activity is required for ocular angiogenesis and inflammation and that reducing the activity of Ref-1 will prevent ocular angiogenesis. Guided by exciting preliminary data, the hypothesis will be tested via two specific aims: Aim 1. Determine the Ref-1-modulated signaling pathway(s) that are key to angiogenesis and inflammation. The expression of Ref-1 in neovascularization will be assessed, and the expression and function of downstream targets (including newly identified Wnt pathway components) will be analyzed after knockdown and inhibition of this protein in endothelial cells with or without overexpression of functional mutants. Angiogenic activity will also be assessed. Aim 2. Optimize the preclinical profile of Ref-1 inhibitors in vitro and in vivo. Two novel, highly potent Ref-1 small molecule inhibitors will be explored for efficacy in vitro, and in cell and in multiple in vivo models of neovascularization, including synergy with anti-vascular endothelial growth factor therapy, target engagement and off-target effects, effects on Ref-1 target genes, and toxicity. This work is innovative, as it is the first in-depth mechanistic study of the role of Ref-1 in ocular angiogenesis, exploring this unique signaling node as an integrator of proangiogenic, proinflammatory, and newly identified Wnt signals. It will also reveal new signaling pathways relevant to angiogenesis and inflammation in the eye, and novel therapeutic leads for neovascular eye diseases. The work is highly significant because it will define Ref-1 as an ocular angiogenic mediator and determine its downstream effects, leading to development of new ways to prevent blindness. Additionally, outcomes from these studies will be the advancement of novel, anti-Ref-1 small molecule inhibitors for translation from the bench to the clinic and patient care.

新生血管性眼病、增殖性糖尿病视网膜病变和湿性年龄相关性黄斑变性是因此，迫切需要寻找新的细胞成分。 Ref-1 蛋白就是这样的成分之一，负责激活对血管生成很重要的氧化还原依赖性转录因子，并过度表达用新型小分子抑制 Ref-1 的氧化还原功能可阻止细胞增殖。激光诱导脉络膜新生血管（L-CNV）模型中的体外和体内眼内皮细胞。 Ref-1 抑制可减少通过缺氧和炎症途径的信号传导，初步数据揭示了与 Wnt 信号传导的新颖联系长期目标是阐明 Ref-1 在眼部的作用。新生血管形成，并开发针对该蛋白质或其途径的新疗法。研究表明，Ref-1 是血管生成和炎症的重要介质，是多种疾病的靶点抗血管生成小分子，以及包括缺氧诱导因子在内的关键血管生成因子的调节剂 1α 和 NF-κB 本应用的目的是确定 Ref-1 如何发挥调节作用。总体假设是 Ref-1 活性。是眼部血管生成和炎症所必需的，降低 Ref-1 的活性将预防眼部血管生成和炎症反应。在令人兴奋的初步数据的指导下，该假设将通过两个具体目标进行检验：目标 1。确定对血管生成和炎症至关重要的 Ref-1 调节信号通路。将评估Ref-1在新生血管形成中的表达，以及下游下游的表达和功能目标（包括新鉴定的 Wnt 通路成分）将在敲低和抑制后进行分析这种蛋白在内皮细胞中过度表达或不过度表达都会产生血管生成活性。目标 2. 优化 Ref-1 抑制剂的体外和体内临床前特性。将探索高效 Ref-1 小分子抑制剂的体外、细胞内和多种研究中的功效。新血管形成的体内模型，包括与抗血管内皮生长因子治疗的协同作用，靶点参与和脱靶效应、对 Ref-1 靶基因的影响以及毒性。因为这是首次对 Ref-1 在眼部血管生成中的作用进行深入的机制研究，探索了这一独特的机制信号转导节点作为促血管生成、促炎症和新发现的 Wnt 信号的整合者。揭示与眼部血管生成和炎症相关的新信号通路以及治疗新药这项工作非常重要，因为它将 Ref-1 定义为眼部疾病。血管生成介质并确定其下游效应，从而开发预防新方法此外，这些研究的成果将是新型抗 Ref-1 小分子药物的进步。分子抑制剂从实验室转移到临床和患者护理。