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 终生致盲的主要原因。因此，迫切需要找到新的细胞成分 这可能是为了阻止眼部新生血管。蛋白质Ref-1就是这样的成分之一， 负责激活对血管生成至关重要的氧化还原依赖的转录因子，并过表达 在新生血管方面。新型小分子抑制Ref-1‘S氧化还原功能阻断细胞增殖 眼内皮细胞在激光诱导的脉络膜新生血管模型中的体内外研究。 抑制REF-1通过缺氧和炎症途径减少信号转导，初步数据显示 与WNT信号的新连接。长期目标是阐明Ref-1在眼球中的作用 新血管形成，并开发针对该蛋白或其途径的新疗法(S)。这样做的理由是 研究表明，Ref-1是血管生成和炎症的重要介质，是多个 抗血管生成小分子，以及包括低氧诱导因子在内的关键血管生成因子的调节 1α和NF-κB.本申请的目标是确定REF-1作为调节器的功能 血管生成，并开发针对这种酶的新试剂。总体假设是Ref-1活性 是眼血管生成和炎症所必需的，降低Ref-1的活性将阻止眼球 血管生成。在令人兴奋的初步数据的指导下，该假说将通过两个具体的目标进行检验：目标1。 确定Ref-1调节的信号通路(S)，这是血管生成和炎症的关键。这个 将评估Ref-1在新生血管中的表达，以及下游基因的表达和功能 靶点(包括新发现的Wnt途径成分)将在击倒和抑制后进行分析 这种蛋白在内皮细胞中有或没有过表达功能突变体。血管生成活性将 也要接受评估。目的2.优化Ref-1抑制剂的体内外临床前图谱。两本小说， 将探索高效的Ref-1小分子抑制剂在体外、细胞内和多种情况下的疗效。 新生血管的活体模型，包括与抗血管内皮生长因子治疗的协同作用， 靶参与和靶外效应，对Ref-1靶基因的影响，以及毒性。这项工作是创新的， 由于这是第一次深入研究Ref-1在眼血管生成中的作用机制，因此探索这一独特的 信号节点作为促血管生成、促炎和新发现的Wnt信号的整合者。它还将 揭示与眼部血管生成和炎症相关的新信号通路，以及新的治疗方法 新生血管眼病的线索。这项工作意义重大，因为它将把Ref-1定义为眼睛 血管生成介体及其下游作用的确定，导致开发新的预防方法 失明。此外，这些研究的结果将是新的、抗REF-1小分子的进步 将分子抑制剂从替补席转移到临床和病人护理。