Epigenetic Control of Retinal Development

视网膜发育的表观遗传控制

• 批准号: 9243446
• 负责人: Rajesh C. Rao
• 金额: $ 22.19万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-01 至 2020-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9243446
• 关键词: Address; Adult; Affect; Age related macular degeneration; Alpha Cell; American; Antibodies; Binding; Biological Assay; Blindness; Caring; Cell Differentiation process; Cell Therapy; Cells; ChIP-seq; Chromatin; Clinical Trials; DNA Sequence; Derivation procedure; Development; Development Plans; Diabetic Retinopathy; Disease; Enhancers; Environment; Enzymes; Epigenetic Process; Faculty; Flow Cytometry; Fostering; Gene Expression Profile; Generations; Genes; Genetic; Genetic Transcription; Genomics; Glaucoma; Goals; Hepatocyte; Histones; Homeodomain Proteins; Human; Impairment; Individual; Inherited; International; Knock-out; Knowledge; Lead; Learning; Luciferases; MLL gene; Mentors; Mentorship; Methods; Michigan; Microphthalmos; Natural regeneration; Nerve Degeneration; Neurons; Organism; Pathway interactions; Photoreceptors; Physicians; Pluripotent Stem Cells; Principal Investigator; Process; Productivity; Program Development; Proliferating; Proteins; Protocols documentation; Public Health; Quality of life; RNA Interference; Repression; Research; Research Personnel; Retina; Retinal; Retinal Degeneration; Retinal Diseases; Retinal Dystrophy; Role; SECTM1 gene; Scientist; Somatic Cell; Stem cells; Surgeon; Techniques; Technology; Testing; Time; Tissues; Training; Transcriptional Regulation; Transplantation; Universities; Vision; Work; Zebrafish; base; career; career development; cell type; chromatin immunoprecipitation; chromatin modification; clinically relevant; design; epigenetic regulation; experimental study; fetal; genome-wide; improved; inhibitor/antagonist; innovative technologies; insight; loss of function; nerve stem cell; novel; regenerative; regenerative therapy; retinal neuron; retinal progenitor cell; skills; transcriptome; transcriptome sequencing; translational scientist; tumor

ABSTRACT: My ultimate goal is to elucidate the epigenetic mechanisms that control retinal development in order to identify novel targets to regenerate tissues lost in the retinal degenerative diseases. These blinding disorders have few treatments and afflict the majority of individuals I care for as a retinal physician and surgeon. For example, age-related macular degeneration (AMD), in which photoreceptors degenerate leading to vision loss, is a disease that affects 10 million Americans. One regenerative strategy for AMD is pluripotent stem cell (PSC)-derived photoreceptor transplantation. However, derivation of PSC-derived retinal progenitor cells (RPCs)—tissue-specific precursors to photoreceptors—remains inefficient, donor-dependent, and poorly understood. To better understand how RPCs arise, and to achieve the initial steps of my career goals, I have planned a three-year, mentored career development program designed to foster my transition to an independent investigator. I have devised didactic and hands-on training aims that integrate stem cells and chromatin dynamics with state-of-the-art epigenetic techniques. This work will take place at University of Michigan (U-M), an outstanding environment with a track record of nurturing young faculty toward independent research careers. My primary mentor, Yali Dou, is an internationally recognized expert in the field of epigenetic regulation of transcription. Sally Temple, co-mentor, is a pioneer in neural stem cells and in defining their developmental transcriptome. Thomas Gardner, U-M K12 Principal Investigator and senior clinician-scientist, whose work focuses on diabetic retinopathy, will serve as a career mentor. This diverse and accomplished team will foster my training aims, career goals, and my efforts to address the central research theme: how an epigenetic enzyme, Mll1, orchestrates formation of RPCs from PSCs. The rationale for this proposal is that by determining the role of Mll1 in retinal differentiation, knowledge will be gained about unknown epigenetic mechanisms that govern retinal formation. We have recently discovered the Mll1-Rx retinal developmental axis, and have also found that Mll1 regulates retinal Meis1. Dysregulation of Mll1, Meis1, or Rx disrupts mammalian retinal development, but how this occurs remains unknown. To address this knowledge gap, the proposed research aims to: 1) determine whether Mll1 deficiency impairs generation of RPCs via Meis1 and Rx repression; and 2) define Mll1-dependent transcriptome and enhancer networks within the retinal Mll1-Rx axis. To accomplish these aims, I will learn and apply innovative technologies such as gene editing, RNA interference, ChIP- and RNA-sequencing, and enhancer motif analysis. This integrated pipeline will allow genome-wide interrogation of targetable and clinically relevant epigenetic pathways in retinal development, and could ultimately be applied to retinal disease. Together, the insights, skills, and guidance gained from the proposed studies, career development plan, and mentorship team will facilitate my transition from a mentored clinician investigator to an independent, R01-supported, translational scientist.

摘要：我的最终目标是阐明控制视网膜发育的表观遗传机制， 以确定新的靶点，使视网膜变性疾病中丢失的组织再生。这些致盲 作为一名视网膜医生，我所治疗的大多数人都患有这种疾病， 外科医生例如，年龄相关性黄斑变性（AMD），其中光感受器退化导致视网膜病变。 视力丧失是一种影响1000万美国人的疾病。AMD的一种再生策略是多能性 干细胞（PSC）衍生的感光细胞移植。然而，PSC衍生的视网膜祖细胞的衍生 细胞（RPC）-光感受器的组织特异性前体-仍然效率低下，依赖供体， 明白为了更好地理解RPC是如何产生的，为了实现我职业目标的最初步骤，我 我计划了一个为期三年的职业发展计划，旨在促进我过渡到一个 独立调查员我设计了教学和动手训练的目标，整合干细胞， 染色质动力学与最先进的表观遗传学技术。这项工作将发生在大学 密歇根大学（U-M），一个杰出的环境与培养年轻教师走向独立的记录 研究生涯。我的主要导师窦雅莉是表观遗传学领域国际公认的专家 转录调控。Sally Temple，共同导师，是神经干细胞的先驱，并在定义其功能方面， 发育转录组托马斯加德纳，U-M K12首席研究员和高级临床科学家， 其工作重点是糖尿病视网膜病变，将作为一个职业导师。这种多样化和成就 团队将促进我的培训目标，职业目标，以及我努力解决中心研究主题：如何 表观遗传酶Mll 1协调PSC形成RPC。提出这项建议的理由是， 通过确定Mll 1在视网膜分化中的作用，将获得关于未知表观遗传的知识。 控制视网膜形成的机制我们最近发现了Mll 1-Rx视网膜发育 轴，并且还发现Mll 1调节视网膜Meis 1。Mll 1、Meis 1或Rx的失调破坏了 哺乳动物视网膜发育，但这是如何发生的仍然未知。为了弥补这一知识差距， 提出的研究旨在：1）确定Mll 1缺陷是否损害经由Meis 1的RPC的产生， Rx抑制;和2）定义视网膜Mll 1-Rx内的Mll 1依赖性转录组和增强子网络 轴线为了实现这些目标，我将学习和应用创新技术，如基因编辑，RNA 干扰、ChIP和RNA测序以及增强子基序分析。这条综合管道将允许 视网膜发育中靶向和临床相关表观遗传途径的全基因组询问，以及 最终可以应用于视网膜疾病。在一起，洞察力，技能，并从获得指导， 建议的研究，职业发展计划和指导团队将有助于我从一个指导的过渡 临床研究者转变为独立的、R 01支持的转化科学家。