Stimulating Retina Regeneration from Muller Cells in Progressive Retinal Degenerations

刺激进行性视网膜变性中 Muller 细胞的视网膜再生

基本信息

批准号：
10379368
负责人：
Brian D Perkins
金额：
$ 51.01万
依托单位：
CLEVELAND CLINIC LERNER COM-CWRU
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10379368
关键词：
1 year old Acute Adult Affect Architecture Bardet-Biedl Syndrome Blindness Cell Death Cell Nucleus Cells Cessation of life Chemicals Chronic Cone Data Degenerative Disorder Disease Disease model Environment Event Exhibits Exposure to Fishes Functional disorder Genes Goals Growth Factor Human Immune Immunohistochemistry Individual Inflammation Inflammatory Inherited Injury Knowledge Leber&apos s amaurosis Light Literature Mammals Mechanics Methods Microglia Modeling Muller&apos s cell Multipotent Stem Cells Mutation Natural regeneration Neurons Pathway interactions Patients Persons Pharmacology Photoreceptors Play Process Prodrugs Proliferating Property Published Comment Quantitative Reverse Transcriptase PCR Regenerative response Reporter Retina Retinal Cone Retinal Degeneration Retinal Diseases Retinal Dystrophy Retinitis Pigmentosa Rod Role Signal Pathway Signal Transduction Source System Testing Therapeutic Time Toxic effect Transgenic Organisms Translating War Zebrafish ciliopathy cytokine cytotoxicity density genetic approach human model immunoregulation inherited retinal degeneration macrophage mouse model mutant neuron loss novel novel strategies photoreceptor degeneration precursor cell progenitor regeneration potential response restoration retinal damage retinal neuron retinal progenitor cell retinal regeneration retinal rods sight restoration stem cells tissue repair transcription factor transcriptome transcriptome sequencing transcriptomics virtual

项目摘要

Inherited retinal degenerations (IRDs) result in the progressive and permanent death of neurons. In recent years, however, our knowledge of endogenous stem cells in the retina has opened the possibility of stimulating regeneration of lost neurons in patients suffering from IRDs. Leber's Congenital Amaurosis (LCA), Bardet-Biedl Syndrome, and Retinitis Pigmentosa are among the most common form of IRDs. Mutations in CEP290 are one of the most common causes of LCA, while mutations in BBS2 contribute to BBS and mutations in the EYS gene cause RP. We demonstrate that zebrafish with mutations in the either cep290, bbs2, or eys genes undergo a progressive cone degeneration with evidence of rod dysfunction. Unlike mammals, zebrafish have the innate ability to regenerate retinal neurons when they are damaged or lost. In response to retinal injury, zebrafish exhibit a robust capability of regenerating lost neurons, including photoreceptors. Retinal damage causes release of growth factors and inflammatory cytokines that trigger Müller glia to divide and generate multipotent retinal progenitor cells that regenerate lost neurons. Central to this process is a reprogramming event that involves activation of Stat3. While robust regeneration occurs following acute injury, evidence from the literature and preliminary data indicate that regeneration does not occur in zebrafish with inherited forms of retinal degeneration, such as the cep290-/-, bbs2-/-, or eys-/- mutants. These observations suggest that Müller glia respond differently to acute vs. inherited forms of retinal injury and that regeneration is only triggered when the degree of retinal injury crosses a “damage threshold” within a temporal window. In zebrafish degeneration mutants there is widespread proliferation of rod progenitor cells. However, Müller glia, which are the source of cone progenitors, fail to proliferate. Using what is known about mechanisms involved in regeneration after acute retinal injury in zebrafish, we will test components of these signaling pathways to determine if they are activated in Müller cells of these photoreceptor degeneration mutants. In particular, we will focus on pathways that converge to activate Stat3. Using RNAseq on purified Muller glia from these degeneration mutants, we will investigate how degeneration and inflammation alter the transcriptome of Muller glia. Finally, we will use pharmacological and genetic approaches to suppress immune cell activity and understand the role of microglia and macrophages on degeneration and regeneration. Understanding the mechanisms that underpin retinal regeneration in multiple zebrafish disease models will generate novel hypotheses that can ultimately be translated into humans with retinal degenerative diseases.

遗传性视网膜变性（IRD）会导致神经元进行性和永久性死亡。然而，近年来，我们对视网膜内源性干细胞的了解开启了刺激患有 IRD 的患者丢失的神经元再生的可能性。先天性黑蒙 (LCA)、Bardet-Biedl 综合征和视网膜色素变性是最常见的疾病 CEP290 突变是 LCA 最常见的原因之一。我们证明，BBS2 的突变会导致 BBS，EYS 基因的突变会导致 RP。 cep290、bbs2 或 eys 基因突变的斑马鱼会经历渐进锥体与哺乳动物不同，斑马鱼具有天生的能力。当视网膜神经元受损或丢失时，斑马鱼会再生视网膜神经元以应对视网膜损伤。表现出再生丢失的神经元（包括视网膜损伤）的强大能力。导致生长因子和炎症细胞因子的释放，触发米勒神经胶质细胞分裂和产生多能视网膜祖细胞，使丢失的神经元再生，这一过程的核心是涉及 Stat3 激活的重编程事件随后发生。急性损伤，文献证据和初步数据表明再生并不发生在具有遗传性视网膜变性的斑马鱼中，例如 cep290-/-、bbs2-/- 或 eys-/- 突变体。这些观察结果表明，穆勒神经胶质细胞对急性和遗传性视网膜变性的反应不同。视网膜损伤的程度，只有当视网膜损伤的程度超过一定程度时才会触发再生在斑马鱼退化突变体中，存在时间窗口内的“损伤阈值”。然而，视杆细胞祖细胞广泛增殖，而穆勒胶质细胞是视锥细胞的来源。利用已知的参与再生的机制，祖细胞无法增殖。斑马鱼急性视网膜损伤，我们将测试这些信号通路的组成部分以确定是否特别是，我们将在这些光感受器变性突变体的穆勒细胞中激活它们。重点关注汇聚以激活 Stat3 的途径，对来自这些细胞的纯化 Muller 胶质细胞使用 RNAseq。变性突变体，我们将研究变性和炎症如何改变转录组最后，我们将使用药理学和遗传学方法来抑制免疫细胞。活动并了解小胶质细胞和巨噬细胞在变性和再生中的作用。了解多种斑马鱼疾病中视网膜再生的机制模型将产生新的假设，最终可以转化为具有视网膜的人类退行性疾病。