Elucidating the role of DNA methyltransferases in epigenetic regulation of retinal regeneration in zebrafish

阐明 DNA 甲基转移酶在斑马鱼视网膜再生表观遗传调控中的作用

• 批准号: 10602467
• 负责人: Ashley Kramer
• 金额: $ 5.27万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10602467
• 关键词: Adult; Affect; American; Anatomy; CRISPR/Cas technology; Candidate Disease Gene; Cell Cycle; Cell Differentiation process; Cell physiology; Cells; Cessation of life; Characteristics; Cicatrix; Communities; Comparative Biology; DNA; DNA Methylation; DNA Modification Methylases; Data; Development; Disease; Down-Regulation; Economic Burden; Epigenetic Process; Exhibits; Funding; Future; Gene Expression; Gene Expression Profile; Genes; Genetic; Gliosis; Goals; Government; Hematopoietic stem cells; Hour; In Vitro; Individual; Injury; Investigation; Knock-out; Literature; Maintenance; Mammals; Mediating; Methods; Modeling; Modification; Molecular; Morphology; Muller' s cell; Multipotent Stem Cells; National Eye Institute; Natural regeneration; Neuroglia; Neurons; Organism; Outcome; Pathway interactions; Phenotype; Process; Regenerative Medicine; Research; Retina; Retinal Diseases; Role; Sequence Homology; Signal Pathway; Signal Transduction; Stem Cell Development; System; Techniques; Testing; Therapeutic; Time; Tissues; Translating; WNT Signaling Pathway; Zebrafish; adult stem cell; beta catenin; cell growth regulation; cell type; demethylation; epigenetic regulation; experimental study; in vivo; in vivo evaluation; knock-down; pluripotency; progenitor; promoter; regenerative; repaired; response; retinal damage; retinal neuron; retinal regeneration; stem cell proliferation; stem cells; success; targeted treatment; teleost; timeline; tissue regeneration; tool; transcription factor

Project Abstract According to the National Eye Institute, there are currently over 12 million Americans suffering from diseases affecting the retina. To present, therapeutic attempts to reduce retinal death or replace lost neurons have been met with limited success, highlighting the need for an alternative approach to this problem. The zebrafish is becoming an increasingly popular model to study mechanisms of stem-cell based tissue regeneration. In response to extensive retinal injury, zebrafish are able to completely regenerate the retina. This is accomplished through the induction of Müller glial cells which undergo an asymmetric division to generate a pool of progenitor cells that go on to regenerate all cell types of the retina with no evidence of the glial scarring observed in mammalian species. While the remarkable capacity of the zebrafish to regenerate tissues has been known for years, our understanding of the comparative biology between mammalian and teleost responses to retinal damage remains poorly understood. To present, studies of retinal regeneration in the zebrafish have largely focused on identifying signaling pathways and individual genes involved in retinal regeneration. The epigenetic orchestration of these pathways, however, remains to be investigated in the zebrafish. DNA methyltransferases (Dnmts) have been studied extensively in mammals and have been identified as critical in the homeostatic maintenance of adult stem cell processes such as hematopoietic stem cell development. The limited literature of epigenetic regulators in zebrafish reveals that teleost species appear to utilize the same mechanisms of epigenetic regulation as mammalian species, with zebrafish Dnmts baring considerable sequence homology to mammalian Dnmts. It is known that immediately following retinal injury in the zebrafish there is an initial global DNA hypomethylation and subsequent increase in DNA methylation as the pools of progenitor cells begin to differentiate, indicating the importance of the epigenetic landscape during these processes. With the experiments planned in this proposal, we aim to identify the role of Dnmts in orchestrating the process of adult retinal regeneration in the zebrafish, with the goal of elucidating pathways regulated in Müller glial derived progenitor cells at the epigenetic level. We also plan to develop a new tool that will be an important addition to the zebrafish community for investigation of the consequences of targeted epigenetic modulation of specific genes. These proposed studies will illuminate candidate genes for targeted therapeutic approaches to identify and “unlock” pathways in mammalian systems that are epigenetically silenced, opening new avenues for future studies in the field of regenerative medicine.

项目摘要 根据国家眼科研究所的数据，目前有超过1200万美国人 来自影响视网膜的疾病。 替换失去的神经元的成功率有限，强调了对敏捷的需求 解决这个问题的方法。 基于干细胞的组织的机制。 斑马鱼能够使视网膜重新生成。 Müller神经胶质的发生，该神经胶质发生不对称分裂以产生祖细胞池 在没有观察到神经胶质疤痕的迹象的情况下，它会发生扭转的所有细胞类型 在哺乳动物中。 多年来，我们对哺乳动物和哺乳动物之间的比较养殖的理解 对视网膜损害损害的反应仍然是众所周知的。 斑马鱼中的再生有大量专注于识别信号通路和 参与视网膜再生的个体基因。 但是，在斑马鱼中仍有待调查。 在哺乳动物中进行了广泛的研究，并在体内稳定中被认为至关重要 维持成年干细胞过程，例如造血干细胞的发育 斑马鱼Revurys的表观遗传经常定常规者的文献有限 具有斑马鱼DNMT的表观遗传调节的机制与哺乳动物物种相同 与哺乳动物DNMT相当的序列同源。 斑马鱼视网膜损伤后，RE是初始的全局DNA降压甲基化和 随后随着祖细胞的池开始分化，DNA甲基化的增加， 在这些过程中指示表观遗传景观 在本提案中计划的实验，我们旨在确定DNMT在策划您中的作用 斑马鱼中成人视网膜再生的过程，目的是Elucidathways 在Müller神经胶质衍生的祖细胞中，我们还计划发展。 斑马鱼社区将是一个重要补充的新工具，以调查 这些提出的研究的靶向表观遗传调节的后果将 针对有针对性的治疗方法照亮候选基因，以识别和途径 在表观遗传沉默的Mamalian系统中，为将来的研究开辟了新的途径 在再生医学领域。