Comparative transcriptomic and epigenomic analyses of Muller glia reprogramming

项目总结： 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 来自内源性视网膜细胞群的穆勒胶质细胞。发掘……的潜力 最终刺激受损的人类视网膜中常驻的Müler神经胶质细胞，我们将采取 使用斑马鱼(再生能力强)、雏鸟(再生能力有限)、 和MICE(再生难)。我们将进行全面和公正的， 视网膜组织中基因表达和染色质构象的比较分析 斑马鱼、雏鸟和小鼠视网膜发育中的祖细胞和Müler胶质细胞。我们还将 研究在三种模式生物被激活/重新编程时改变它们的Müler神经胶质细胞 对视网膜损伤(光损伤，NMDA)或暴露于以下外部因素的反应 能够在没有视网膜损伤的情况下诱导它们的激活。目标1和目标2将生成 相关基因和染色质结构的转录组和染色质数据 Müler神经胶质前体细胞的形成。在目标3中，我们将使用新的 发展生物信息学分析以确定转录因子和转录网络 控制每个生物体的Müler神经胶质细胞的神经生成能力。在目标4中，我们将验证 并测试调节斑马鱼、雏鸟和小鼠Müler胶质细胞去分化的候选基因 小鼠，采用功能获得和功能丧失相结合的方法。这项工作将开始 确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。了解如何让穆勒·格里亚恢复青春 状态将使有针对性的再生视网膜治疗成为可能。

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.