Hippo Signaling in Regeneration of the Retinal Pigment Epithelium

视网膜色素上皮再生中的河马信号传导

基本信息

批准号：
10596550
负责人：
Sara Ramirez
金额：
$ 0.7万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10596550
关键词：
11 cis Retinal Address Adult Age related macular degeneration Atrophic Binding Sites Biological Assay Blindness Cell Cycle Cell Nucleus Cell Proliferation Cells Chemicals Choroid Chronic Disease DNA Binding Data Development Developmental Biology Disease Elderly Electroretinography Embryo Eye Eye diseases Functional disorder Future Gene Expression Genes Genetic Genetic Enhancer Element Genetic Transcription Goals Growth Homeostasis Human Immunofluorescence Immunologic In Situ In Vitro Injury Knowledge Ligation Luciferases Maintenance Molecular Mus Natural regeneration Neuroepithelial Neurofibromin 2 Nonexudative age-related macular degeneration Nuclear Nuclear Translocation Optical Coherence Tomography Pathway interactions Phagocytosis Phosphorylation Population Proliferating Proliferation Marker Promoter Regions Property Recycling Regenerative response Retina Retinal Degeneration Retinal Photoreceptors Role Signal Pathway Signal Transduction Specific qualifier value Structure of retinal pigment epithelium Technology Testing Therapeutic Time Tissues Transcription Coactivator Vision Zebrafish cell fate specification cell regeneration chromatin immunoprecipitation epithelial injury epithelium regeneration experimental study genetic manipulation in vivo in vivo regeneration inducible Cre injury and repair monolayer novel postmitotic prevent regenerative regenerative biology response response to injury retinal regeneration sodium iodate transcription factor transdifferentiation wound healing

项目摘要

PROJECT SUMMARY The retinal pigment epithelium (RPE) is an essential monolayer that lies between the retina and the choroid and supports the function and integrity of the retinal photoreceptors. The RPE maintains tissue homeostasis in the adult mainly via long-term survival instead of cell turnover. Degeneration and atrophy of the central RPE causes the detrimental chronic disease age-related macular degeneration (AMD), which is the leading cause of blindness in the elderly population. A possibility for treating dry AMD in the future is by stimulating endogenous RPE regeneration, but little is known about the mechanisms that can drive RPE regeneration in vivo. One candidate pathway is the Hippo signaling pathway, as it plays a role in regulating cell proliferation and regeneration in many tissues and is also required in the developing RPE for cell fate specification. In developmental studies, we and others have discovered that Neurofibromin 2 (NF2) is critical for balancing RPE growth and proliferation. My preliminary data suggests that modulation of Hippo signaling through Nf2 disruption in the adult stimulates proliferation and differentiation of the RPE following injury. This proposal aims to determine how genetic manipulation of Hippo signaling promotes an intrinsic proliferative and regenerative response in the mammalian RPE. In Aim 1, I will characterize the role of NF2 in promoting RPE regeneration after injury in the adult. I have established an injury paradigm, in which I chemically injure the RPE of mice. Regeneration will be assessed on a cellular level by expression, co-localization, and quantification of RPE and proliferative markers, and on a functional level by electroretinography. In Aim 2, I will determine the mechanism by which YAP regulates RPE specification and maintenance in the RPE. Several studies demonstrate that YAP is required for RPE identity in the embryo and adult; however, the molecular mechanism is not well understood. Here, I will investigate the hypothesis that YAP and the transcription factor TEAD directly regulate expression of RPE- specific genes by using chromatin immunoprecipitation and luciferase assays. The results of these experiments will reveal how YAP regulates RPE specification, and how the Hippo signaling pathway can be modulated to promote RPE regeneration. Understanding the cellular and molecular mechanisms that promote RPE regeneration will help identify and develop therapeutic strategies for AMD.

项目概要视网膜色素上皮 (RPE) 是位于视网膜和脉络膜之间的重要单层细胞，支持视网膜感光器的功能和完整性。 RPE 维持组织稳态成体主要通过长期存活而不是细胞更替。中央 RPE 变性和萎缩的原因有害的慢性疾病与年龄相关的黄斑变性（AMD），这是导致黄斑变性的主要原因老年人口失明。未来治疗干性 AMD 的一种可能性是通过刺激内源性 RPE 再生，但对于体内驱动 RPE 再生的机制知之甚少。一候选途径是Hippo信号途径，因为它在调节细胞增殖和许多组织的再生，并且在开发用于细胞命运规范的 RPE 时也是必需的。在发育研究中，我们和其他人发现神经纤维蛋白 2 (NF2) 对于平衡 RPE 至关重要生长和增殖。我的初步数据表明，通过破坏 Nf2 来调节 Hippo 信号在成人中，损伤后刺激 RPE 的增殖和分化。该提案旨在确定 Hippo 信号传导的基因操作如何促进细胞内的内在增殖和再生反应哺乳动物 RPE。在目标 1 中，我将描述 NF2 在促进损伤后 RPE 再生中的作用成人。我建立了一种损伤范例，用化学方法损伤小鼠的 RPE。再生将是通过 RPE 和增殖标记物的表达、共定位和定量在细胞水平上进行评估，并通过视网膜电图在功能水平上进行。在目标 2 中，我将确定 YAP 的调节机制 RPE 规范和 RPE 维护。多项研究表明 RPE 需要 YAP 胚胎和成人的同一性；然而，其分子机制尚不清楚。在这里，我将研究 YAP 和转录因子 TEAD 直接调节 RPE-表达的假设通过使用染色质免疫沉淀和荧光素酶测定来确定特定基因。这些实验的结果将揭示 YAP 如何调节 RPE 规范，以及如何调节 Hippo 信号通路促进RPE再生。了解促进 RPE 的细胞和分子机制再生将有助于确定和制定 AMD 的治疗策略。