Role of Runx1 in Pathologic Ocular Angiogenesis

Runx1 在病理性眼血管生成中的作用

• 批准号: 10165721
• 负责人: LEO A KIM
• 金额: $ 52.81万
• 依托单位: SCHEPENS EYE RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-05 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10165721
• 关键词: Affect; Age; Age related macular degeneration; Area; Atrophic; Binding Sites; Bioinformatics; Blindness; Blood Vessels; Cell physiology; Choroidal Neovascularization; Chronic; Clinical; Data; Defect; Development; Disease; Elderly; Endothelial Cells; Exudative age-related macular degeneration; Eye; Eye diseases; Gene Expression Profile; Gene Targeting; Genes; Genetic; Genetic Models; Genetic Transcription; Glucose; Human; In Vitro; Infant; Injections; Lasers; Lead; Light Coagulation; Mediating; Mediator of activation protein; Membrane; Methodology; Methods; Modality; Modeling; Molecular; Mus; Ontology; Oxygen; Pathologic; Pathologic Neovascularization; Pathway interactions; Patients; Physiologic Neovascularization; Physiological; Premature Infant; Process; Proteins; Publishing; Research; Retina; Retinal Diseases; Retinal Neovascularization; Retinopathy of Prematurity; Risk; Role; Sequence Analysis; Testing; Therapeutic; Therapeutic Agents; Tissues; Transcriptional Regulation; Treatment Factor; United States; Validation; Vascular Endothelial Cell; Vascular Endothelial Growth Factors; Vision; acute myeloid leukemia 1 protein; angiogenesis; base; bioinformatics tool; demographics; differential expression; experimental study; gain of function; in vivo; knock-down; loss of function; man; migration; mouse model; neovascularization; novel; ocular angiogenesis; ocular neovascularization; overexpression; patient subsets; postnatal; postnatal development; preclinical efficacy; proliferative diabetic retinopathy; retina blood vessel structure; screening; therapeutic target; transcription factor; transcriptome; transcriptome sequencing

PROJECT SUMMARY Pathologic ocular angiogenesis is the underlying mechanism of a variety of sight threatening diseases of the eye affecting a wide range of patients including premature infants and the elderly. Retinopathy of prematurity (ROP), proliferative diabetic retinopathy (PDR), and exudative age-related macular degeneration (wet AMD) are common causes of blindness in infants, working age, and the elderly respectively. These conditions are primarily characterized by aberrant neovascularization, or the formation of vascular membranes on top or below the retina. We performed transcriptome analysis of vascular endothelial cells (ECs) obtained from fibrovascular membranes (FVMs) from patients with PDR, and identified Runt-related transcription factor 1 (Runx1) as a gene that is upregulated in pathologic ocular angiogenesis. We subsequently showed that Runx1 mediated EC function, and that its inhibition leads to a reduction in aberrant angiogenesis in a mouse model of oxygen-induced retinopathy (OIR). Based on these data, we hypothesize that Runx1-mediated angiogenesis is an important mechanism for aberrant angiogenesis within the eye, and that Runx1 inhibition is a potential therapeutic modality for the treatment of a wide array of pathologic angiogenic diseases. To test this hypothesis, the following research aims are proposed: Aim 1. To determine the role of Runx1 in physiological and pathological angiogenesis using genetic models of Runx1 loss- and gain-of-function: Using inducible models of Runx1 loss- and gain-of-function (LOF/GOF), we will evaluate the role of Runx1 in both physiological and pathological angiogenesis. We will look at postnatal development of the retinal vessels, as well as pathologic angiogenesis using laser-induced choroidal neovascularization (CNV). Aim 2. To determine the downstream effectors of Runx1 transcriptional activity. Using transcriptome analysis, we propose to identify genes that are potentially regulated by Runx1 using human retinal microvascular endothelial cells and Runx1 LOF/GOF models. Using a combination of bioinformatics screening methodologies (gene ontology analysis, Runx1 regulatory sequence analysis, and comparison to our published human PDR transcriptomes) and in vitro validation of gene targets, we propose to identify the direct/indirect downstream mediators that regulate Runx1-mediated angiogenesis. Aim 3. To compare anti-Runx1 versus anti-VEGF treatment in genetic and inducible models of pathologic ocular angiogenesis. Anti-Runx1 therapy has potential benefits over anti-VEGF therapy. In this aim, we propose to compare anti-Runx1 versus anti-VEGF therapy alone or in combination using multiple models of angiogenesis: OIR, Akimba, and laser-induced CNV. Thus this aim will elucidate the translational potential of anti-Runx1 therapy for the management of such diseases as ROP, PDR, and wet AMD.

项目摘要 病理性眼部血管生成是多种视力威胁性疾病的潜在机制， 影响包括早产儿和老年人在内的广泛患者的眼睛。早产儿视网膜病变 (ROP)、增殖性糖尿病视网膜病变（PDR）和渗出性年龄相关性黄斑变性（湿性AMD） 分别是婴儿、工作年龄和老年人失明的常见原因。这些条件 主要特征为异常的新血管形成，或在顶部或顶部形成血管膜， 在视网膜下面。我们进行了血管内皮细胞（EC）的转录组分析，这些细胞来自于 纤维血管膜（FVM）的PDR患者，并确定Runt相关转录因子1 （Runx1）作为在病理性眼部血管生成中上调的基因。我们随后发现Runx1 介导的EC功能，并且其抑制导致小鼠模型中异常血管生成的减少。 氧诱导视网膜病变（OIR）。基于这些数据，我们假设Runx1介导的血管生成是一个重要的机制。 这是眼内异常血管生成的重要机制，Runx1抑制是一种潜在的 用于治疗多种病理性血管生成疾病的治疗方式。为了验证这一 假设，提出以下研究目标： 目标1.为了确定Runx1在生理和病理性血管生成中的作用，使用遗传学方法， Runx1功能丧失和获得模型：使用Runx1功能丧失和获得的诱导模型 (LOF/GOF），我们将评估Runx1在生理和病理血管生成中的作用。我们将 看看视网膜血管的出生后发育，以及使用激光诱导的病理性血管生成， 脉络膜新生血管（CNV）。 目标2.确定Runx1转录活性的下游效应子。使用转录组 分析，我们建议确定基因，可能是由Runx1调控使用人类视网膜 微血管内皮细胞和Runx1 LOF/GOF模型。结合生物信息学筛选 方法（基因本体分析，Runx1调控序列分析，并与我们发表的 人PDR转录组）和基因靶点的体外验证，我们建议鉴定直接/间接 调节Runx1介导的血管生成的下游介质。 目标3。比较抗Runx1与抗VEGF治疗在遗传性和诱导性肿瘤模型中的作用， 病理性眼部血管生成抗Runx1疗法比抗VEGF疗法具有潜在益处。在这 目的是，我们建议使用多种抗Runx1与抗VEGF治疗进行比较， 血管生成模型：OIR、Akimba和激光诱导的CNV。因此，这一目标将阐明翻译 抗Runx1治疗在ROP、PDR和湿性AMD等疾病管理中的潜力。