Comparative transcriptomic and epigenomic analyses of Muller glia reprogramming

Muller 胶质细胞重编程的比较转录组和表观基因组分析

• 批准号: 9551199
• 负责人: John D Ash
• 金额: $ 57.62万
• 依托单位: UNIVERSITY OF NOTRE DAME
• 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/9551199
• 关键词: ATAC-seq; Animal Model; Bioinformatics; Birds; Blindness; Candidate Disease Gene; Cell Differentiation process; Cells; ChIP-seq; Chromatin; Chromatin Structure; Competence; DNA; Data; Development; Exposure to; Fishes; Gene Expression; Gene Expression Profiling; Gene Structure; Genes; Genetic Transcription; Growth Factor; Human; In Situ; In Situ Hybridization; Individual; Injury; Light; Mammals; Messenger RNA; MicroRNAs; Modeling; Molecular; Molecular Conformation; Muller' s cell; Mus; N-Methylaspartate; Natural regeneration; Neurons; Organism; Pathway interactions; Population; Primates; Proliferating; Property; Quantitative Reverse Transcriptase PCR; Refractory; Regulator Genes; Rest; Retina; Retinal; Rodent; Sampling; Signal Pathway; Societies; Stem cells; Stimulus; Testing; Transcriptional Regulation; Treatment Factor; United States; Validation; Vision; Visual Acuity; Visual impairment; Work; Zebrafish; blind; comparative; design; disability; epigenomics; gain of function; in vivo; infancy; loss of function; neurogenesis; novel therapeutics; overexpression; prevent; progenitor; regenerative; response; retinal damage; retinal neuron; retinal progenitor cell; retinal regeneration; transcription factor; transcriptome; transcriptome sequencing; transcriptomics; virtual

Project Summary: One potentially important approach to restore vision is the regeneration of lost retinal neurons from an endogenous population of retinal cells, the Müller glia. To explore the potential of ultimately stimulating the resident Müller glia in the damaged human retina, we will take a comparative approach using zebrafish (regeneration competent), chick (regeneration limited), and mouse (regeneration refractory). We will conduct a comprehensive and unbiased, comparative analysis of gene expression and chromatin conformation in isolated retinal progenitor cells and Müller glia in developing zebrafish, chick, and mouse retinas. We will also study changes Müller glia from all three model organisms as they are activated/reprogrammed in response to retinal injury (light damage, NMDA) or exposure to extrinsic factors that are capable of inducing their activation in the absence of retinal damage. Aims 1 and 2 will generate transcriptome and chromatin data of genes and chromatin structures that are associated with formation of Müller glia progenitor cells. In Aim 3, we will integrate this data using newly developed bioinformatic analysis to identify transcription factors and transcriptional networks that control neurogenic competence in Müller glia from each organism. In Aim 4, we will validate and test candidate genes in regulating the dedifferentiation of Müller glia in zebrafish, chick, and mice, using a combination of gain- and loss-of-function approaches. This work will begin to identify the transcription factors and miRNAs that regulate the extent of retinal regeneration in the three different model organisms. Understanding how to restore Müller glia to a youthful status will enable targeted regenerative retinal therapies.

项目概要： 恢复视力的一种潜在重要方法是丢失的视网膜神经元的再生 来自内源性视网膜细胞群，即米勒神经胶质细胞。探索潜力 最终刺激受损人类视网膜中的穆勒神经胶质细胞，我们将采取 使用斑马鱼（具有再生能力）、雏鸡（再生能力有限）的比较方法， 和小鼠（再生难治性）。我们将进行全面、公正的调查， 离体视网膜基因表达和染色质构象的比较分析 斑马鱼、小鸡和小鼠视网膜发育过程中的祖细胞和米勒神经胶质细胞。我们还将 研究改变了所有三种模式生物的米勒神经胶质细胞，因为它们被激活/重新编程 响应视网膜损伤（光损伤，NMDA）或暴露于以下外在因素 能够在没有视网膜损伤的情况下诱导其激活。目标 1 和 2 将生成 与相关基因和染色质结构的转录组和染色质数据 米勒神经胶质祖细胞的形成。在目标 3 中，我们将使用新的方法来整合这些数据 开发生物信息分析来识别转录因子和转录网络 控制每个生物体穆勒神经胶质细胞的神经发生能力。在目标 4 中，我们将验证 并测试调节斑马鱼、雏鸡和小鼠 Müller 胶质细胞去分化的候选基因 小鼠，使用功能获得和功能丧失相结合的方法。这项工作将开始 鉴定调节视网膜再生程度的转录因子和 miRNA 三种不同的模式生物。了解如何使穆勒胶质细胞恢复年轻 状态将使有针对性的视网膜再生治疗成为可能。