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

• 批准号: 10385720
• 负责人: Sara Ramirez
• 金额: $ 3.15万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 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/10385720
• 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; Equilibrium; Eye; 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; Mitotic; Molecular; Mus; Natural regeneration; Neuroepithelial; Neurofibromin 2; Nonexudative age-related macular degeneration; Nuclear; Nuclear Translocation; Optical Coherence Tomography; Pathway interactions; Phagocytosis; Play; Population; Proliferating; Proliferation Marker; Promoter Regions; Property; Recycling; Regenerative response; Retina; Retinal Degeneration; Retinal Photoreceptors; Role; Signal Pathway; Signal Transduction; 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; injury and repair; monolayer; novel; prevent; regenerative; regenerative biology; response; response to injury; retinal regeneration; sodium iodate; transcription factor; transdifferentiation; wound healing

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.

项目摘要 视网膜色素上皮（RPE）是一个必不可少的单层，位于视网膜和脉络膜之间 支持视网膜光感受器的功能和完整性。 RPE维持组织稳态 成人主要是通过长期生存而不是细胞更新。中央RPE原因的退化和萎缩 有害的慢性疾病年龄相关的黄斑变性（AMD），这是 老年人口的失明。将来处理干燥AMD的可能性是刺激内源性 RPE再生，但对可以在体内驱动RPE再生的机制知之甚少。一 候选途径是河马信号通路，因为它在调节细胞增殖和 在许多组织中的再生，在开发RPE中也需要用于细胞命运规范。在 发展研究，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NF2破坏对河马信号的调节 在成年人中，刺激受伤后RPE的增殖和分化。该建议旨在确定 河马信号传导的遗传操纵如何促进固有的增殖和再生反应 哺乳动物RPE。在AIM 1中，我将表征NF2在促进RPE在受伤后促进RPE再生中的作用 成人。我已经建立了损伤范式，其中我化学损伤了小鼠的RPE。再生将是 通过表达，共定位和RPE和增殖标记的定量在细胞水平上评估， 以及在功能层面上通过电子模拟层面。在AIM 2中，我将确定YAP调节的机制 RPE中的RPE规范和维护。几项研究表明RPE需要YAP 胚胎和成人的身份；但是，分子机制尚不清楚。在这里，我会的 研究YAP和转录因子TEAD直接调节RPE-表达的假设 使用染色质免疫沉淀和荧光素酶测定法。这些实验的结果 将揭示YAP如何调节RPE规范，以及如何调节河马信号通路 促进RPE再生。了解促进RPE的细胞和分子机制 再生将有助于识别和制定AMD的治疗策略。