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： 恢复视力的一个潜在的重要方法是再生丢失的视网膜神经元 来自视网膜细胞的内源性群体，缪勒神经胶质。为了探索 最终刺激受损人类视网膜中的穆勒神经胶质，我们将采取一种 使用斑马鱼（再生能力），小鸡（再生有限）， 和小鼠（再生难治性）。我们将进行全面和公正的， 视网膜色素上皮细胞基因表达和染色质构象的比较分析 祖细胞和Müller神经胶质细胞在发育中的斑马鱼，小鸡和小鼠视网膜。我们还将 研究改变了所有三种模式生物的Müller胶质细胞，因为它们被激活/重编程 对视网膜损伤（光损伤，NMDA）或暴露于 能够在没有视网膜损伤的情况下诱导它们的激活。目标1和2将产生 基因和染色质结构的转录组和染色质数据， Müller神经胶质祖细胞的形成。在目标3中，我们将使用新的 开发了生物信息学分析，以识别转录因子和转录网络 控制每种生物体的缪勒神经胶质的神经原能力。在目标4中，我们将验证 并测试候选基因在调节斑马鱼，鸡， 小鼠，使用功能获得和功能丧失方法的组合。这项工作将开始， 鉴定调节视网膜再生程度的转录因子和miRNAs， 三种不同的模式生物了解如何恢复穆勒神经胶质到一个年轻的 这将使有针对性的视网膜再生治疗成为可能。