Testing Gene Therapy in Models of Geographic Atrophy

在地理萎缩模型中测试基因疗法

基本信息

批准号：
10011817
负责人：
Alfred S Lewin
金额：
$ 52.3万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-08-01 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10011817
关键词：
AHR gene Acute Adenoviruses Age Age related macular degeneration American Animal Model Animals Anti-Inflammatory Agents Antioxidants Aryl Hydrocarbon Receptor Astrocytes Atrophic Biodistribution Blindness Bruch&apos s basal membrane structure CASP1 gene Cells Cessation of life Characteristics Cholesterol Homeostasis Choroidal Neovascularization Chronic Collaborations Complement Dependovirus Deposition Diagnosis Disease Disease model Elderly Enzymes Exhibits Exudative age-related macular degeneration Eye Functional disorder Gene Transduction Agent Gene Transfer Genes Goals Human Hypertrophy Inflammasome Inflammation Inflammatory Inflammatory Response Injections Interleukin-1 beta Investigational Drugs Lead Lipofuscin Macular degeneration Measurement Methods Microglia Mitochondria Modeling Muller&apos s cell Mus Neural Retina Nonexudative age-related macular degeneration Optical Coherence Tomography Outcome Oxidants Oxidative Stress Pathology Pathway interactions Patients Peptides Pharmaceutical Preparations Photography Photoreceptors Process Production Proteins Research Project Grants Retina Retinal Diseases Retinal Pigments Risk SOD2 gene Signal Transduction Signaling Molecule Site Structure of retinal pigment epithelium Technology Testing Therapeutic Tight Junctions Time Tissues Toxicology Toxin Translational Research Viral Vector Virus Vision adeno-associated viral vector advanced disease age related antioxidant enzyme cell type cigarette smoke cytokine design effective therapy gene therapy geographic atrophy human disease insight intravitreal injection macula mouse model oxidative damage prevent promoter retinal damage subretinal injection therapeutic gene vector

项目摘要

Chronic inflammation is a characteristic, and possibly a cause, of the retinal damage associated with age related macular degeneration (AMD), a leading cause of blindness among the elderly. Inflammation may be a consequence of age related oxidative stress or accumulation of toxins within the RPE and retina. This is a project to develop gene therapy for AMD by preventing inflammatory processes signaled by potent cytokines such as IL-1β. It is our hypothesis that, by reducing inflammation associated with these signaling molecules, we can halt the progression of AMD in patients diagnosed with early stage disease. There are two forms of the advanced disease, wet (exudative) AMD and geographic atrophy, both of which lead to loss of central vision. While there are effective treatments for wet AMD, there is no treatment for geographic atrophy, which leads to death of the retinal pigment epithelium and photoreceptor cells in the macula. Furthermore, atrophic processes may persist in wet AMD, even when choroidal neovascularization is under control. This project is a collaboration between two groups that have generated distinct mouse models for geographic atrophy. One of the models increases oxidative stress in the retinal pigment epithelium (RPE) by deleting the gene for a protective enzyme, manganese superoxide dismutase. The second model deletes the gene for the aryl hydrocarbon receptor (AhR), causing ocular dysregulation of several pathways relevant to AMD including cholesterol homeostasis and clearance of oxidative stress-inducing toxins such as those found in cigarette smoke. Both models result in key features of dry AMD including accumulation of basal deposits in the RPE, damage to Bruch's membrane, RPE atrophy and dysfunction and death of photoreceptors. We propose to develop gene therapy in these models using adeno-associated virus (AAV) which has been demonstrated to be safe for gene transfer to the retina. Importantly, this virus does not provoke a severe inflammatory response. Our approach to therapy in the two models is to deliver genes for secreted, cell penetrating proteins that increase protection from oxidative stress or that reduce the production of inflammatory cytokines by stimulating synthesis of anti-oxidant enzymes or blocking the activity of the NLRP3 inflammasome and preventing the activation of caspase-1. We will validate that delivery of the secreted anti- inflammatory proteins to the vitreous produces peptides that permeate the retina and RPE. In the final component of the project, we will use cell-type specific viral vectors that produce non-secreted peptides in order to identify the cells that are major producers of inflammatory cytokines in our mouse models. Our goal is to develop gene therapy vectors that could be tested in patients, and we believe that one-time delivery of a therapeutic vector to the vitreous will be translatable to human gene therapy.

慢性炎症是与年龄相关的残留损害的特征，也是可能的原因相关的黄斑变性（AMD），这是老年失明的主要原因。炎症可能是与年龄相关的氧化物应激或RPE和视网膜内毒素积累的结果。这是一个通过预防有效细胞因子签署的炎症过程来开发AMD的基因疗法的项目例如IL-1β。我们的假设是，通过减少与这些信号分子相关的炎症，我们可以停止在诊断出患有早期疾病的患者中AMD的进展。有两种形式的晚期疾病，湿（渗出性）AMD和地理萎缩，这两者都导致中央视力丧失。虽然有有效的湿AMD治疗方法，但没有用于地理萎缩的治疗方法，这导致黄斑中视网膜色素上皮和感光细胞的死亡。此外，萎缩过程即使在控制脉络膜新血管形成的情况下，也可能会持续存在。该项目是两组之间的合作，它们为地理生成了不同的鼠标模型萎缩。其中一种模型通过删除视网膜色素上皮（RPE）的氧化应激。用于保护酶，锰超氧化物歧化酶的基因。第二个模型删除了基因芳基烃受体（AHR），导致与AMD相关的几种途径的眼部失调胆固醇稳态和氧化应激诱导的毒素的清除，例如在香烟中发现的毒素抽烟。这两种模型都均可均可取得干燥AMD的关键特征，包括加速RPE中的基本沉积物，损害Bruch的膜，RPE萎缩和功能障碍以及光感受器的死亡。我们建议使用腺相关病毒（AAV）在这些模型中开发基因治疗证明对基因转移到视网膜是安全的。重要的是，该病毒不会引起严重的炎症反应。我们在这两个模型中的治疗方法是为分泌的细胞提供基因穿透性蛋白质会增加免受氧化应激的保护或减少产生的蛋白质通过刺激抗氧化酶的合成或阻断NLRP3活性来炎症细胞因子炎症体并阻止caspase-1的激活。我们将验证分泌的抗对玻璃体的炎性蛋白会产生渗透视网膜和RPE的肽。在决赛中项目的组成部分，我们将使用细胞类型的特定病毒载体，这些病毒载体在为了鉴定小鼠模型中炎症细胞因子的主要产生者的细胞。我们的目标是要开发可以在患者中测试的基因疗法载体，我们认为一次性递送玻璃体的治疗载体将被转换为人类基因疗法。