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

• 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
• 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

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 失明的主要原因。因此，迫切需要找到新的细胞成分 可以靶向阻断眼部新生血管。蛋白质Ref-1就是这样一种组分， 负责激活对血管生成重要的氧化还原依赖性转录因子，并过表达 新生血管形成用新的小分子抑制Ref-1的氧化还原功能阻断了细胞增殖， 激光诱导的脉络膜新生血管（L-CNV）模型中的眼内皮细胞的体外和体内研究。 Ref-1抑制通过缺氧和炎症途径减少信号传导，初步数据显示， 与Wnt信号的新联系。长期的目标是阐明Ref-1在眼神经系统中的作用。 新血管形成，并开发靶向该蛋白质或其途径的新疗法。这样做的理由 研究表明，Ref-1是血管生成和炎症的重要介质，是多种肿瘤的靶点， 抗血管生成小分子和关键血管生成因子的调节因子，包括缺氧诱导因子 1α和NF-κB。本申请的目的是确定Ref-1如何作为 血管生成，并开发针对这种酶的新药物。总体假设是Ref-1活性 是眼部血管生成和炎症所必需的，并且降低Ref-1的活性将防止眼部血管生成和炎症。 血管生成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测试：目标1。 确定Ref-1调节的信号通路是血管生成和炎症的关键。的 将评估Ref-1在新血管形成中的表达，并且将评估下游Ref-1的表达和功能。 靶点（包括新鉴定的Wnt途径组分）将在敲低和抑制后进行分析。 这种蛋白在内皮细胞中有或没有过表达的功能突变体。血管生成活性将 也要进行评估。目标2.优化Ref-1抑制剂的体外和体内临床前特征。两部小说， 将探索高效Ref-1小分子抑制剂在体外、细胞和多个细胞中的功效， 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， 靶向接合和脱靶效应、对Ref-1靶基因的效应和毒性。这项工作是创新的， 因为这是第一个深入研究Ref-1在眼部血管生成中作用的机制，探索这种独特的 信号传导节点作为促血管生成、促炎症和新鉴定的Wnt信号的整合者。它还将 揭示了与眼内血管生成和炎症相关的新信号通路， 导致新生血管性眼病。这项工作非常重要，因为它将把Ref-1定义为目镜， 血管生成介质，并确定其下游的影响，导致开发新的方法，以防止 失明此外，这些研究的结果将是新的抗Ref-1小分子的进展。 分子抑制剂，从实验室到临床和患者护理。