DNA Demethylation and Muller Glia Reprogramming During Retina Regeneration

视网膜再生过程中 DNA 去甲基化和米勒胶质细胞重编程

基本信息

批准号：
8502787
负责人：
DANIEL J GOLDMAN
金额：
$ 18.84万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-04-01 至 2015-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8502787
关键词：
Animals Antisense Oligonucleotides Automobile Driving Birds Blindness Cell physiology Cells Code CpG dinucleotide DNA DNA Methylation DNA Sequence Deaminase Development Disease Epigenetic Process Exhibits Eye Injuries Eye diseases Failure Fishes Fluorescence-Activated Cell Sorting Gene Expression Gene Expression Profiling Gene Silencing Generations Genes Genetic Materials Genetic Programming Genome Genomics Histones Human Inherited Injury Libraries Location Mammals Methylation Microarray Analysis Multipotent Stem Cells Natural regeneration Neuroglia Neurons Organism Population Process Proliferating Proteins Retina Retinal Role Site Somatic Cell System Tail Testing Transcriptional Regulation Transgenic Organisms Variant Vision Work X Inactivation Zebrafish bisulfite cell type demethylation embryonic stem cell imprint induced pluripotent stem cell injured knock-down novel novel strategies prevent programs public health relevance regenerative repaired retinal progenitor cell teleost fish

项目摘要

DESCRIPTION (provided by applicant): Unlike mammals, zebrafish are able to regenerate a damaged retina and restore lost sight. This regenerative ability depends on Muller glia (MG) that respond to retinal injury by undergoing multiple shifts in identity as they dedifferentiate, proliferate, and finally differentiate to regenerate new neurons and glia. Although MG can be coaxed to proliferate in the injured mammalian retina, they do not exhibit multipotency and only rarely regenerate damaged neurons. Therefore understanding the mechanisms driving zebrafish MG reprogramming to mutlipotency may suggest novel strategies for generating multipotent progenitors from mammalian MG. Recent studies suggest that MG reprogramming is accompanied by activation of gene expression programs that are similar to those acting in embryonic stem cells and retinal progenitors. We hypothesize that genetic programs driving MG dedifferentiation and multipotency are controlled by DNA methylation. In animals, DNA methylation predominantly occurs at CpG dinucleotides and controls transcriptional regulatory processes like imprinting, X-chromosome inactivation, transposon silencing, and stable silencing of gene activity. Methylation of DNA proximal to gene-coding regions is correlated with gene silencing. Importantly, changes in DNA methylation have been correlated with the activation and suppression of gene expression programs that take place during early development and accompany the reprogramming of somatic cells to yield induced pluripotent stem cells. It is likely that erasure and reestablishment of genomic methylation, at key locations, accompanies the gene expression changes that drive MG dedifferentiation and multipotency and subsequently the regeneration of new retinal cell types. Here we propose to identify regions of the MG genome that are undergoing methylation changes during retina regeneration and determine if these changes correlate with gene expression changes that have previously been characterized using microarray technology. In addition, we propose to test the hypothesis that DNA demethylation in dedifferentiating MG is an active process driven by Apobec2a and 2b proteins.

描述（由申请人提供）：与哺乳动物不同，斑马鱼能够再生损坏的视网膜并恢复失去的视力。这种再生能力取决于Muller Glia（Mg），这些能力通过在去分化，增殖并最终区分以再生新的神经元和神经胶质的同时，对视网膜损伤做出反应。尽管可以在受伤的哺乳动物视网膜中哄骗MG增殖，但它们不表现出多稳定性，并且很少会再生受损的神经元。因此，了解驱动斑马鱼MG重编程至Mutlipotency的机制可能表明可以从哺乳动物Mg产生多能祖细胞的新策略。最近的研究表明，MG重编程伴随着与在胚胎干细胞和视网膜祖细胞中作用的基因表达程序的激活。我们假设驱动MG去分化和多能力的遗传程序受DNA甲基化控制。在动物中，DNA甲基化主要发生在CpG二核苷酸上，并控制转录调节过程，例如印刷，X染色体灭活，转座子沉默和基因活性的稳定沉默。 DNA近端与基因编码区域的甲基化与基因沉默有关。重要的是，DNA甲基化的变化已与早期发育过程中发生的基因表达程序的激活和抑制相关，并伴随着体细胞的重编程以产生诱导的多能干细胞。在关键位置擦除和重建基因组甲基化的可能是伴随的基因表达变化，驱动Mg去分化和多稳定性，然后再生新的视网膜细胞类型的再生。在这里，我们建议确定在视网膜再生过程中正在经历甲基化变化的MG基因组的区域，并确定这些变化是否与以前使用微阵列技术表征的基因表达变化相关。此外，我们建议测试以下假设：去分化Mg中的DNA去甲基化是由APOBEC2A和2B蛋白驱动的活性过程。