Epigenetic Regulation of Retinal Development by BCOR

BCOR 对视网膜发育的表观遗传调控

• 批准号: 10367943
• 负责人: Michelle Zhang
• 金额: $ 5.18万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10367943
• 关键词: 5' Untranslated Regions; Address; Affect; Anophthalmos; Binding; Cell Differentiation process; Cell Extracts; Cell Line; Cells; ChIP-seq; Childhood; Chromatin; Chromatin Remodeling Factor; Complex; Congenital Disorders; Data; Defect; Dental Pulp; Development; Disease; Embryo; Epigenetic Process; Eye Development; Eye diseases; Frameshift Mutation; Gatekeeping; Gene Mutation; Generations; Genes; Genetic Transcription; Global Change; HDAC1 gene; Histone H2A; Human; Immunofluorescence Immunologic; In Situ Hybridization; Knowledge; LHX2 gene; Link; Lysine; Mass Spectrum Analysis; Mediating; Microdissection; Microphthalmos; Modeling; Modification; Mutate; Mutation; Neurodegenerative Disorders; Optic vesicle; Organoids; PRC1 Protein; Patients; Persons; Phenotype; Play; Polycomb; Process; Proteins; Quality of life; RB1 gene; RNA; Rana; Regenerative Medicine; Regulation; Retina; Retinal Degeneration; Retinal Diseases; Retinoblastoma; Role; Stains; Syndrome; Testing; Time; Tissues; Transcript; Variant; Vision; Visual Fields; Xenopus; Zebrafish; base; cell type; chromatin remodeling; design; epigenetic regulation; experimental study; gene network; histone modification; improved; in vitro Model; in vivo; induced pluripotent stem cell; knock-down; malignant neoplasm of eye; neurogenesis; novel; optic cup; recruit; regenerative approach; retinal progenitor cell; stem cells; transcription factor; transcriptome sequencing

Project Summary Normal vision depends on the balanced generation and homeostatic regulation of major retinal cell types, and the disruption of these processes may result in congenital retinal disorders and retinal degeneration, affecting millions of people around the world each year. Regenerative medicine depends on our understanding of retinal development for informing retinal cell reprogramming and manufacturing. While there have been significant advances in unravelling the roles of transcription factors, a major gap exists in our knowledge of how these tightly regulated gene networks are governed by epigenetic mechanisms. Polycomb-group complexes are evolutionarily conserved epigenetic machineries that remodel chromatin through histone modifications for silencing targeted genes and have been recently shown to affect retinal neurogenesis and lineage decisions. BCOR is a component of the Polycomb Repressive Complex 1 variant that ubiquitinates lysine 119 on histone H2A (H2AK119Ub). Dysregulation of BCOR is clearly associated with eye and retinal disorders, including Lenz microphthalmia, Oculofaciocardiodental Syndrome, and retinoblastoma, but its role in retinal neurogenesis is still not well understood. In preliminary studies, I performed coimmunoprecipitation-mass spectrometry and validated BCOR’s interaction with other PcG proteins in retinal cells. I then performed BCOR chromatin immunoprecipitation sequencing (ChIP-seq) and identified strong peaks at retinal progenitor genes LHX2, PAX6, SIX3, and SIX6, all of which are significant in optic vesicle development and later play distinct roles in retinal cell fate decisions. Thus, I propose to investigate the role of BCOR in retinal neurogenesis. I hypothesize that BCOR mediates the switch from retinal progenitors to differentiated cell types by recruiting PcG proteins to regulate H2AK119Ub on the chromatin to repress multipotency genes. I will test this hypothesis with the following aims: (1) Identify the epigenetic mechanism of BCOR for regulating retinal progenitor genes, and (2) Determine how BCOR affects the generation of major retinal cell types in vivo. My overall objective is to characterize BCOR-ncPRC1.1’s role in retinal development as an avenue to discovering new epigenetic targets for regenerative strategies in the retina.

项目概要 正常视力取决于主要视网膜细胞类型的平衡生成和稳态调节，并且 这些过程的破坏可能导致先天性视网膜疾病和视网膜变性，影响 每年全世界有数百万人。再生医学取决于我们对视网膜的理解 为视网膜细胞重编程和制造提供信息的开发。虽然已经出现了重大 尽管在阐明转录因子的作用方面取得了进展，但我们对这些转录因子如何发挥作用的知识仍存在重大差距。 严格调控的基因网络受到表观遗传机制的控制。多梳族配合物是 进化上保守的表观遗传机制，通过组蛋白修饰重塑染色质 沉默目标基因，最近被证明可以影响视网膜神经发生和谱系决定。 BCOR 是 Polycomb Repressive Complex 1 变体的一个组成部分，可泛素化组蛋白上的赖氨酸 119 H2A (H2AK119Ub)。 BCOR 失调显然与眼睛和视网膜疾病有关，包括 Lenz 小眼症、眼面心牙综合征和视网膜母细胞瘤，但其在视网膜神经发生中的作用是 还是不太理解。在初步研究中，我进行了共免疫沉淀-质谱分析并 验证了 BCOR 与视网膜细胞中其他 PcG 蛋白的相互作用。然后我进行了 BCOR 染色质检测 免疫沉淀测序 (ChIP-seq) 并鉴定出视网膜祖基因 LHX2 的强峰， PAX6、SIX3 和 SIX6，所有这些在视神经泡发育中都具有重要意义，并在后来的 视网膜细胞的命运决定。因此，我建议研究 BCOR 在视网膜神经发生中的作用。我 假设 BCOR 通过募集介导从视网膜祖细胞到分化细胞类型的转变 PcG 蛋白调节染色质上的 H2AK119Ub 以抑制多能基因。我会测试这个 假设具有以下目的：（1）确定BCOR调节视网膜的表观遗传机制 (2) 确定 BCOR 如何影响体内主要视网膜细胞类型的生成。我的 总体目标是描述 BCOR-ncPRC1.1 在视网膜发育中的作用，作为发现的途径 视网膜再生策略的新表观遗传目标。