Novel Activators of Regeneration in Muller glia

穆勒胶质细胞再生的新型激活剂

• 批准号: 9340183
• 负责人: EDWARD M LEVINE
• 金额: $ 63.51万
• 依托单位: VANDERBILT UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9340183
• 关键词: Address; Affinity; Affinity Chromatography; Alpha Cell; Animal Model; Area; Behavior; Biological Models; Cell Cycle Proteins; Cell Separation; Cells; Collaborations; Cyclin-Dependent Kinase Inhibitor; Data; Disease; Eye; Fishes; Genetic; Goals; Human; Injectable; Injury; Intervention; Knowledge; Label; Laboratories; Membrane; Messenger RNA; MicroRNAs; Muller' s cell; Mus; Natural regeneration; Nuclear; Pathology; Pathway interactions; Pharmacology; Photoreceptors; Population; Preparation; Proliferating; Property; Proteins; Proteomics; Protocols documentation; RNA; Reagent; Records; Regenerative response; Reporter; Research; Retina; Retinal; Scientist; Signal Transduction; Source; Stem cells; Techniques; Testing; Therapeutic; Thiouracil; Vesicle; Work; Zebrafish; adult stem cell; base; cell type; comparative; exosome; gabazine; gamma-Aminobutyric Acid; in vivo; innovation; migration; mouse model; neoplastic cell; neurotransmitter uptake; next generation sequencing; novel; novel strategies; prevent; public health relevance; receptor; regenerative; repaired; retinal rods; teleost fish; transcription factor; transcriptome; transcriptome sequencing; transcriptomics

Project Summary: Novel Activators of Regeneration in Müller Glia Müller glia are the predominant source of regenerative repair in the fish retina, but this is not the case in humans, where Müller glia respond to injury, pathology, or degeneration by becoming reactive, and, in many cases, gliotic. Recent research in non-regenerative mammalian model systems, such as the mouse, suggests that Müller glia could be pushed down a regenerative pathway, but bona-fide regeneration and repair will involve factors and protocols that are not yet identified. To address this gap in knowledge, we propose three projects that focus on the proliferative and neurogenic properties needed for regeneration. In the first project, we will develop a regeneration-priming protocol for mouse Müller glia through modulation of neurotransmitter uptake and inhibition of p27Kip1, a cell cycle protein that prevents properties associated with reactivity and regeneration in Müller glia. In the second project, we will perform a screen using exosomes to discover candidate exosomes and their constituent proteins and/or RNAs that stimulate regenerative properties in zebrafish and mouse Müller glia. In the third project, we will develop thiouracil tagging combined with RNA sequencing to capture the transcriptomes of Müller glia in selected states of regeneration based on our work from the first two projects. The culmination of this work will yield new factors for promoting regeneration and provide proof-of-concept for exosome-based delivery of factors as a therapeutic strategy.

项目摘要：穆勒胶质细胞再生的新型激活剂 米勒神经胶质细胞是鱼类视网膜再生修复的主要来源，但在鱼类视网膜中情况并非如此 人类中，米勒神经胶质细胞通过变得反应性来对损伤、病理或退化做出反应，并且在许多情况下 病例, 胶质增生。最近对非再生哺乳动物模型系统（例如小鼠）的研究表明 米勒神经胶质细胞可以被推向再生途径，但真正的再生和修复将 涉及尚未确定的因素和协议。为了解决这一知识差距，我们提出了三个建议 专注于再生所需的增殖和神经源性特性的项目。在第一个项目中， 我们将通过调节神经递质开发小鼠穆勒神经胶质细胞的再生启动方案 p27Kip1 的摄取和抑制，p27Kip1 是一种细胞周期蛋白，可防止与反应性和相关性相关的特性 米勒神经胶质细胞的再生。在第二个项目中，我们将使用外泌体进行筛选以发现 候选外泌体及其组成蛋白和/或RNA刺激再生特性 斑马鱼和小鼠穆勒神经胶质细胞。第三个项目，我们将开发结合RNA的硫尿嘧啶标签 根据我们的工作进行测序以捕获选定再生状态下的穆勒胶质细胞的转录组 从前两个项目来看。这项工作的最终成果将产生促进再生和 为基于外泌体的因子递送作为治疗策略提供概念验证。