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: 10647870
• 负责人: Mark R. Kelley
• 金额: $ 45.92万
• 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 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10647870
• 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; Mission; Modeling; Mus; National Eye Institute; New Agents; Oral; Outcome Study; Oxidation-Reduction; Oxygen; Pathway interactions; Patient Care; Patients; Pharmaceutical Preparations; Phase; Play; Proliferating; 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; innovation; knock-down; migration; mutant; neovascular; neovascularization; new therapeutic target; novel; novel therapeutics; ocular angiogenesis; ocular neovascularization; overexpression; oxidation; pre-clinical; prevent; proliferative diabetic retinopathy; retinal toxicity; small molecule; small molecule inhibitor; success; synergism; systemic toxicity; 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.

新血管疾病可增殖的糖尿病性视网膜病和与年龄相关的黄斑变性 在整个生命周期中失明的主要原因。因此，迫切需要找到新的细胞成分 这可能是针对阻断椭圆形新血管形成的。蛋白质ref-1是这样的组成部分， 负责激活依赖氧化还原的转录因子，对血管生成很重要，并且过表达 用新型小分子抑制Ref-1的氧化还原函数阻碍了 眼部内皮细胞体外和体内激光诱导的脉络膜新生血管形成（L-CNV）模型。 Ref-1抑制作用可通过缺氧和感染途径降低信号传导，初步数据揭示了一个 与Wnt信号传导的新型连接。长期目标是阐明Ref-1在眼中的作用 新生血管形成，并开发针对该蛋白质或其途径的新型疗法。理由 研究是Ref-1是血管生成和炎症的重要介体，一个多重目标 抗血管生成的小分子和关键血管生成因子的调节剂，包括缺氧诱导因子 1α和NF-κB。本应用程序中的对象是确定ref-1如何作为调节器的作用 血管生成并开发针对该酶的新药物。总体假设是Ref-1活动 眼血管生成和炎症需要，减少Ref-1的活性将防止眼 血管生成。在令人兴奋的初步数据的指导下，该假设将通过两个特定目的进行检验：目标1。 确定Ref-1调节的信号通路，这是血管生成和炎症的关键。这 将评估Ref-1在新血管形成中的表达，并将其表达和功能的表达和功能 敲低和抑制后将分析目标（包括新确定的Wnt途径组件） 该蛋白质在内皮细胞中具有或没有功能突变体过表达的蛋白质。血管生成活性会 也可以评估。 AIM 2。在体外和体内优化Ref-1抑制剂的临床前谱。两本小说， 将探索高潜力的Ref-1小分子抑制剂，以在体外效率，在细胞和多个中 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 目标参与和脱靶效应，对REF-1靶基因的影响以及毒性。这项工作是创新的， 因为它是Ref-1在眼血管生成中的作用的第一次深入的机械研究，探索了这种独特的 信号传导节点是促血管生成，促炎和新鉴定的Wnt信号的集成剂。它也会 揭示与眼睛的血管生成和炎症有关的新信号通路和新疗法 引导新血管疾病。这项工作非常重要，因为它将Ref-1定义为眼部 血管生成介质并确定其下游影响，从而发展新方法 失明。此外，这些研究的结果将是新型抗REF-1小的进步 分子抑制剂，用于从长凳上转换为诊所和患者护理。

项目成果

Inhibition of APE1/Ref-1 for Neovascular Eye Diseases: From Biology to Therapy.

• DOI: 10.3390/ijms221910279
• 发表时间: 2021-09-24
• 期刊: International journal of molecular sciences
• 影响因子: 5.6
• 作者: Hartman GD;Lambert-Cheatham NA;Kelley MR;Corson TW
• 通讯作者: Corson TW

An improved method for murine laser-induced choroidal neovascularization lesion quantification from optical coherence tomography images.

• DOI: 10.1016/j.mex.2022.101809
• 发表时间: 2022
• 期刊: METHODSX
• 影响因子: 1.9
• 作者: Jensen, Nathan R.;Lambert-Cheatham, Nathan;Hartman, Gabriella D.;Muniyandi, Anbukkarasi;Park, Bomina;Sishtla, Kamakshi;Corson, Timothy W.
• 通讯作者: Corson, Timothy W.

APE1/Ref-1 as a Novel Target for Retinal Diseases.

• DOI: 10.33696/signaling.2.044
• 发表时间: 2021
• 期刊: Journal of cellular signaling
• 作者: Heisel C;Yousif J;Mijiti M;Charizanis K;Brigell M;Corson TW;Kelley MR
• 通讯作者: Kelley MR

Decreased Expression of Soluble Epoxide Hydrolase Suppresses Murine Choroidal Neovascularization.

• DOI: 10.3390/ijms232415595
• 发表时间: 2022-12-09
• 期刊: International journal of molecular sciences
• 影响因子: 5.6

Cancer Research in the "Chemical Biology" Section of the Journal Molecules.

• DOI: 10.3390/molecules25225275
• 发表时间: 2020-11-12
• 期刊: Molecules (Basel, Switzerland)
• 作者: Corson TW