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 与年龄相关的黄斑变性（AMD）是一种眼神经退行性疾病，说明 大约一半的发达国家的失明和视觉障碍病例。据估计 到2040年，将被诊断出2.88亿人在全球范围内被诊断出。 对于AMD患者的严重视力丧失的大约80％，其特征是在亚a 视网膜空间。视网膜下纤维化划定末期新血管AMD，导致视力丧失，甚至 进行抗VEGF疗法。因此，必须揭示下视网膜下的分子机制 形成纤维化和开发基于药物的疗法，不仅用于黄斑新血管形成（MNV），还针对 预防或治疗视网膜下纤维化。积累的证据表明上皮到间质转变 视网膜上皮细胞（RPE）和内皮到间质转变（ENDOMT）的（EMT） 脉络膜或视网膜内皮细胞均有助于视网膜下纤维化病变中的间充质细胞群。 此外，胚胎的内分鼠会导致内皮屏障的渗透性增加，并可以提供解释 随着时间的推移，抗VEGF治疗的疗效降低。因此，旨在抑制或反向的疗法 EMT和ENDOMT可以稳定内皮屏障功能，并预防或减少视网膜下纤维化。各种各样的 细胞外细胞因子和生长因子与激活MNV，EMT和胚胎有关。我们已经确定了 小型GTPase ARF6作为信号通路中的收敛点，许多这些细胞因子和 增长因素。因此，我们假设ARF6激活诱导MNV，EMT和ENDOMT，从而 有助于视网膜下纤维化的发作，并且对ARF6的遗传丧失或药理抑制作用将 减少新血管AMD中的MNV和视视纤维化。为了检验这一假设，我们将追求三个目标。 在AIM 1中，我们将确定是否需要ARF6激活脉络膜和视网膜下属和视网膜下 激光诱导的CNV和JR5558小鼠模型中的纤维化。我们还将使用激光诱导的CNV模型， 内皮细胞谱系跟踪和单细胞RNASEQ，以确定ARF6在endomt中的作用。在AIM 2中，我们 将确定在激光诱导的CNV中RPE EMT和视网膜下纤维化是否需要ARF6激活 和JR5555鼠标模型，并使用细胞谱系跟踪和单细胞RNASEQ来确定 EMT中的ARF6。在AIM 3中，我们将对EMT和ENDOMT的作用进行增添或协同作用以促进 新生血管AMD以及药理学抑制ARF6是否可以减少小鼠的新血管AMD 型号。这项研究可能会通过鉴定ARF6来对新血管AMD的未来治疗产生重大影响 作为药物开发的潜在目标。此外，这项研究也可能对 治疗其他眼血管疾病，例如增生性玻璃体病，前膜和 纤维血管疾病中的视网膜纤维化，例如增生性糖尿病性视网膜病和早产性视网膜病变。