Inhibiting Neovascularization and Subretinal Fibrosis in Neovascular Age-Related Macular Degeneration

抑制新生血管性年龄相关性黄斑变性的新生血管形成和视网膜下纤维化

• 批准号: 10639785
• 负责人: Mary Elizabeth Ruth Hartnett
• 金额: $ 62.94万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10639785
• 关键词: Accounting; Adult; Age related macular degeneration; Blindness; Blood Vessels; Bruch' s basal membrane structure; Cell Culture Techniques; Cell Lineage; Cells; Choroid; Cicatrix; Data; Degenerative Disorder; Developed Countries; Development; Diagnosis; Disease; Drug Targeting; Endothelial Cells; Endothelium; Epiretinal Membrane; Epithelial Cells; Exudative age-related macular degeneration; Fibroblast Growth Factor; Fibrosis; Future; Genetic; Growth; Growth Factor; Human; Inflammatory; Interleukin-1; Interleukin-1 beta; Knock-out; Knockout Mice; Lasers; Lesion; Location; Mesenchymal; Modeling; Molecular; Monomeric GTP-Binding Proteins; Myofibroblast; Neurodegenerative Disorders; Patients; Permeability; Persons; Pharmaceutical Preparations; Pigment Epithelium; Population; Production; Proliferative Vitreoretinopathy; Proteins; Publishing; Retina; Retinal Neovascularization; Retinopathy of Prematurity; Role; Signal Pathway; Structure of retinal pigment epithelium; TNF gene; Testing; Time; Transforming Growth Factor beta; Treatment Factor; Vascular Diseases; Vascular Endothelial Cell; Vascular Endothelial Growth Factors; Vision; Visual; cell motility; connective tissue growth factor; cytokine; disability; drug development; epithelial to mesenchymal transition; experimental study; extracellular; inhibitor; macula; mouse model; neovascular; neovascularization; novel therapeutics; pharmacologic; prevent; proliferative diabetic retinopathy; protein function; response; single-cell RNA sequencing; therapy design; wound healing

PROJECT SUMMARY Age-related macular degeneration (AMD) is an ocular neurodegenerative disorder that accounts for approximately half of all cases of blindness and visual disability in developed nations. It is estimated that around 288 million people will be diagnosed with AMD worldwide by 2040. The neovascular form of AMD, accounting for about 80% of the severe vision loss in patients with AMD, is characterized by neovascularization in the sub- retinal space. Subretinal fibrosis demarcates end-stage neovascular AMD, causing irreparable vision loss, even with anti-VEGF therapy. Thus, it is imperative to unravel the molecular mechanisms underpinning subretinal fibrosis formation and develop drug-based therapies not only for macular neovascularization (MNV) but also to prevent or treat subretinal fibrosis. Accumulating evidence suggests that epithelial-to-mesenchymal transition (EMT) of the retinal epithelial cell (RPE) and endothelial-to-mesenchymal transition (EndoMT) of either the choroidal or retinal endothelial cells all contribute to the mesenchymal cell population in subretinal fibrotic lesions. Additionally, EndoMT leads to an increase in endothelial barrier permeability and could provide an explanation for the reduced efficacy of anti-VEGF treatment over time. Therefore, therapies designed to inhibit or reverse EMT and EndoMT could stabilize endothelial barrier function and prevent or reduce subretinal fibrosis. Various extracellular cytokines and growth factors are involved in activating MNV, EMT, and EndoMT. We have identified the small GTPase ARF6 as a convergence point in signaling pathways activated by many of these cytokines and growth factors. Thus, we hypothesize that ARF6 activation induces MNV, EMT, and EndoMT, thereby contributing to the onset of subretinal fibrosis, and that genetic loss or pharmacological inhibition of ARF6 will reduce both MNV and subretinal fibrosis in neovascular AMD. To test this hypothesis, we will pursue three aims. In Aim 1, we will determine whether ARF6 activation is required for choroidal and retinal EndoMT and subretinal fibrosis in laser-induced CNV and JR5558 mouse models. We will also use the laser-induced CNV model, endothelial cell lineage tracing, and single cell RNAseq to determine the role of ARF6 in EndoMT. In Aim 2, we will determine whether ARF6 activation is required for RPE EMT and subretinal fibrosis in laser-induced CNV and JR5558 mouse models and use cell lineage tracing and single cell RNAseq to determine the function of ARF6 in EMT. In Aim 3, we will investigate whether EMT and EndoMT act additively or synergistically to promote neovascular AMD and whether pharmacologic inhibition of ARF6 can reduce neovascular AMD in mouse models. This study could have a major impact on the future treatment of neovascular AMD by identifying ARF6 as a potential target for drug development. Moreover, this study could also have important implications for the treatment of other ocular vascular diseases, e.g., proliferative vitreoretinopathy, epiretinal membranes, and retinal fibrosis in fibrovascular diseases, such as proliferative diabetic retinopathy and retinopathy of prematurity.

项目摘要 视网膜相关性黄斑变性（AMD）是一种眼部神经退行性疾病， 在发达国家，大约一半的失明和视力残疾病例。据估计， 到2040年，全球将有2.88亿人被诊断患有AMD。AMD的新生血管形式，占 对于约80%的AMD患者的严重视力丧失，其特征在于亚- 视网膜间隙视网膜下纤维化界定了终末期新生血管性AMD，导致不可挽回的视力丧失，甚至 抗VEGF治疗。因此，有必要解开视网膜下的分子机制， 纤维化形成，并开发基于药物的治疗方法，不仅用于黄斑新生血管（MNV）， 预防或治疗视网膜下纤维化。越来越多的证据表明上皮细胞向间质细胞的转化 (EMT)视网膜上皮细胞（RPE）和内皮-间充质转化（EndoMT） 脉络膜或视网膜内皮细胞都对视网膜下纤维化病变中的间充质细胞群有贡献。 此外，EndoMT导致内皮屏障通透性增加，并可能提供一种解释， 随着时间的推移，抗VEGF治疗的功效会降低。因此，旨在抑制或逆转 EMT和EndoMT可稳定内皮屏障功能，预防或减轻视网膜下纤维化。各种 细胞外细胞因子和生长因子参与激活MNV、EMT和EndoMT。我们已经确定 小的GT3/ARF 6作为由许多这些细胞因子激活的信号传导途径中的汇聚点， 生长因子因此，我们假设ARF 6激活诱导MNV、EMT和EndoMT， 导致视网膜下纤维化的发生，并且ARF 6的遗传缺失或药物抑制将 减少新生血管性AMD中的MNV和视网膜下纤维化。为了验证这一假设，我们将追求三个目标。 在目标1中，我们将确定脉络膜和视网膜EndoMT和视网膜下EndoMT是否需要ARF 6活化。 激光诱导的CNV和JR 5558小鼠模型中的纤维化。我们还将使用激光诱导的CNV模型， 内皮细胞谱系追踪和单细胞RNAseq，以确定ARF 6在EndoMT中的作用。在目标2中， 将确定激光诱导的CNV中RPE EMT和视网膜下纤维化是否需要ARF 6激活 和JR 5558小鼠模型，并使用细胞谱系追踪和单细胞RNAseq来确定 急诊室的急性呼吸衰竭6型。在目标3中，我们将研究EMT和EndoMT是否相加或协同作用，以促进 新生血管性AMD和药物抑制ARF 6是否可以减少小鼠的新生血管性AMD 模型这项研究通过鉴定ARF 6，可能对未来新生血管性AMD的治疗产生重大影响。 作为药物开发的潜在目标。此外，这项研究也可能对 治疗其它眼血管疾病，例如，增殖性玻璃体视网膜病变、视网膜前膜，和 纤维血管疾病中的视网膜纤维化，例如增殖性糖尿病视网膜病变和早产儿视网膜病变。