Epigenetic basis of retinal cell fate determination

视网膜细胞命运决定的表观遗传学基础

基本信息

批准号：
10627931
负责人：
Issam Al Diri
金额：
$ 39.13万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10627931
关键词：
3-Dimensional ATOH7 gene Affect Architecture Binding Binding Sites Biological Assay CRISPR/Cas technology Cell Differentiation process Cells ChIP-seq Chromatin Complex DNA Data Degenerative Disorder Development Differentiation and Growth Disease Elements Enhancers Epigenetic Process Essential Genes Expression Profiling Eye diseases Foundations Gene Activation Gene Expression Generations Genes Genetic Genetic Enhancer Element Genome Genome Mappings Genomic DNA Genomics Glaucoma Growth Hi-C Human In Vitro Injury Knowledge Link Maintenance Maps Molecular Mus Neuropathy Nuclear Optic Nerve Pathway interactions Patients Persons Population Proliferating Regulation Regulatory Element Reporter Genes Resolution Retina Retinal Degeneration Retinal Ganglion Cells Role SOX11 gene SOX4 gene Specific qualifier value Techniques Technology Testing Therapeutic Transcriptional Regulation Transgenic Organisms Transplantation Untranslated RNA Visual impairment Work YY1 Transcription Factor Zebrafish axon growth cell replacement therapy chromosome conformation capture experimental study ganglion cell gene regulatory network genome wide association study in vivo innovation insight next generation sequencing promoter retinal ganglion cell degeneration retinogenesis stem cells tool transcription factor

项目摘要

SUMMARY: Progressive loss of retinal ganglion cells (RGCs) and subsequent degeneration of optic nerve is among the most common ophthalmic neuropathies that affect populations worldwide. A promising therapeutic strategy relies on using stem cells to reconstruct functional retinal ganglion cells in vitro that can be used to replace dying cells in affected patients. However, for this strategy to be successful, a thorough understanding to the molecular mechanisms of RGC specification and differentiation are needed to elucidate and exploit normal RGC developmental pathways and thereby maximize RGC generation for cell replacement therapies. Our previous work has mapped the epigenetic landscape dynamics during mouse and human developing retina. However, there is a fundamental gap in our knowledge to the role of 3D chromatin topology in RGC development and maintenance. Experiments in this proposal will address this role by combining innovative state of the art genomic and genetic tools to elucidate the chromatin architecture dynamics that accompany RGC genesis and to determine how they function in vivo. Our central hypothesis is that temporal and spatial regulation of ganglion cell genesis is associated with dynamic 3D genomic interactions between specific non-coding DNA elements and genes that drive RGC differentiation and optic nerve growth. To test this hypothesis, we will elucidate the link between 3D chromatin architecture and the expression of a transcription factor essential for RGC formation. We will also dissect the functional significance of constituents of the regulatory landscape that accommodates RGC differentiation in vivo. Finally, we will integrate ChIP-Seq data with techniques that map chromatin topology to elucidate the long-range genomic interactions that are associated with components of the regulatory networks that are required for RGC differentiation. When completed the results of this proposal will advance our understanding to the 3D regulatory landscape that accommodates the generation of RGCs in vivo, a necessary knowledge to enhance RGC replacement therapies in RGC degenerative diseases. Project Summary- Al Diri 1

摘要：视网膜神经节细胞（RGC）的进行性丧失和随后的变性视神经是影响种群的最常见的眼科神经病全世界。有希望的治疗策略依赖于使用干细胞重建功能视网膜神经节细胞体外可用于替代受影响患者的垂死细胞。然而，为了使该策略成功，对RGC的分子机制有透彻的理解需要规格和差异化来阐明和利用正常的RGC发展途径，从而使RGC最大化用于细胞替代疗法。我们的先前工作绘制了鼠标和人类发展过程中的表观遗传景观动态视网膜。但是，我们的知识对3D染色质拓扑的作用存在根本差距在RGC开发和维护中。该提案中的实验将通过结合艺术基因组和遗传工具的创新状态以阐明染色质伴随RGC创世纪并确定它们在体内的功能的架构动力学。我们的中心假设是，神经节细胞发生的时间和空间调节是与特定非编码DNA元件之间的动态3D基因组相互作用相关以及驱动RGC分化和视神经生长的基因。为了检验这一假设，我们将阐明3D染色质体系结构与转录因子的表达之间的链接 RGC形成必不可少的。我们还将剖析成分的功能意义适用于体内RGC分化的监管景观。最后，我们将整合用绘制染色质拓扑结构的技术来阐明远程基因组与监管网络的组件相关的交互 RGC分化。完成后，该提案的结果将提高我们的理解到适合体内RGC的3D监管景观，这是必要的知识以增强RGC退行性疾病中的RGC替代疗法。项目摘要diri 1