Epigenomic mechanisms regulating RGC survival and axon regeneration

调节 RGC 存活和轴突再生的表观基因组机制

• 批准号: 10318187
• 负责人: Feng Tian
• 金额: $ 10.17万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2023-03-27
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10318187
• 关键词: ATAC-seq; Accounting; Address; Affect; Animal Model; Architecture; Axon; Biological Assay; Blindness; Boston; Brain; CCCTC-binding factor; CRISPR/Cas technology; Cell Death; Cell Survival; Chromatin; Chromatin Structure; Clustered Regularly Interspaced Short Palindromic Repeats; Crush Injury; Data; Epigenetic Process; Event; Eye; Failure; Fluorescence-Activated Cell Sorting; Gene Expression; Gene Expression Alteration; Gene Expression Profile; Genes; Genetic Screening; Genetic Transcription; Glaucoma; Individual; Injections; Injury; Institutes; Knowledge; Lead; Light; Maintenance; Maps; Mediating; Mentors; Methodology; Methods; Modeling; Molecular; Molecular Target; Mus; Natural regeneration; Nerve Crush; Neuraxis; Neurobiology; Neurodegenerative Disorders; Neurons; Ophthalmology; Optic Disk; Optic Nerve; Optic Nerve Injuries; Outcome; Pathologic; Patients; Pediatric Hospitals; Persons; Phase; Physiologic Intraocular Pressure; Positioning Attribute; Principal Investigator; Procedures; Proteins; Publishing; Regenerative Medicine; Research; Retina; Retinal Diseases; Retinal Ganglion Cells; Role; Science; Signal Pathway; Silicone Oils; Structure; Technology; Testing; Therapeutic; Time; Training Activity; Transposase; United States; Universities; Virus Diseases; Vision; Work; XCL1 gene; axon regeneration; axonal degeneration; base; career development; cell regeneration; central nervous system injury; chromatin remodeling; effective therapy; epigenetic regulation; epigenome; epigenomics; experimental study; in vivo; injured; insight; knock-down; medical schools; mouse model; neurodevelopment; neuron loss; neuronal survival; new therapeutic target; novel therapeutics; optic nerve disorder; programs; recruit; regenerative; regenerative biology; regenerative therapy; research and development; retinal ganglion cell degeneration; retinal ganglion cell regeneration; retinal neuron; screening; stem cell biology; tool; transcription factor; transcriptome sequencing; visual information

Project Summary/Abstract The proposed study is a five-year career development research plan that focuses on dissecting the epigenetic regulation of retinal ganglion cell (RGC) degeneration and regeneration in mouse models of optic nerve crush and glaucoma. The candidate is currently a postdoctoral research fellow at F.M. Kirby Neurobiology Center at Boston Children’s Hospital and Harvard Medical School. The candidate intends to further extend his expertise in epigenomic profiling technologies, mechanisms of optic neuropathies and development of neural regenerative therapeutics by integrating the mentor team of Dr. Zhigang He at Boston Children’s Hospital and Harvard Medical School, Dr. Joshua Sanes at Harvard University and Harvard Brain Science Initiative, Dr. Jeffery Goldberg at Stanford University and Byers Eye Institute, and Dr. Jason Buenrostro at Harvard Stem Cell and Regenerative Biology (SCRB) Department and Broad Institute of MIT and Harvard. The candidate has also recruited Dr. Daniel Geschwind, a collaborator of his current and proposed studies, as his advisor for specific scientific and technical support. The proposed experiments and training activities will enable the candidate to publish top-tier ophthalmology research works and uniquely position him as an independent principal investigator pursuing novel therapeutics for retinal disease such as glaucoma. Glaucoma is the second leading cause of blindness in the United States with at least 3,000,000 people affected. This number is likely to increase by 60% by 2030 if no new therapeutics could be developped. Due to the inablity of central nervous system to regenerate after injury, the vision loss resulted from RGC death is irreversible and will lead to permanent blindness. Our preliminary experiments using the CRISPR/Cas9-based in vivo forward genetic screen have discovered that the knockdown of CCCTC-binding factor (CTCF) could robustly promote axonal regeneration of RGCs while do not increase overall RGC survival, and that the knockdown of structure specific recognition protein 1 (SSRP1) significantly protected RGC survival without facilitating axon regeneration. To explore the underlying epigenetic mechanisms regulating RGC survival and regeneration, the proposed study will specifically pursue the following aims: (1) To profile injury-induced chromatin remodeling in RGCs by ATAC sequencing (mentored phase); (2) To assess the mechanisms that mediate differential effects of CTCF and SSRP1 upon injury (mentored and independent phase); and (3) To assess the mechanisms and effects of epigenetic regulators in a glaucoma model (independent phase). The outcome of the proposed study will provide in-depth and quantitative insights into why and how the regenerative fates of RGCs are pre-determined from an epigenomic perspective, which can be directly transformed to new cures for optic neuropathies.

项目总结/摘要 这项拟议中的研究是一项为期五年的职业发展研究计划，重点是解剖表观遗传 视神经夹伤小鼠视网膜神经节细胞变性和再生的调控 和青光眼。候选人目前是F.M.的博士后研究员。柯比神经生物学中心 波士顿儿童医院和哈佛医学院。候选人打算进一步扩展他的专业知识 在表观基因组分析技术，视神经病变的机制和神经再生的发展， 通过整合波士顿儿童医院和哈佛医学中心的何志刚博士的导师团队， 学校，哈佛大学的约书亚萨尼斯博士和哈佛脑科学倡议，杰弗里戈德堡博士在 斯坦福大学和拜尔斯眼科研究所，以及哈佛干细胞和再生医学中心的杰森？ 麻省理工学院和哈佛大学生物学系和布罗德研究所。候选人还招募了丹尼尔博士 Geschwind，他目前和拟议研究的合作者，作为他的顾问，具体的科学和技术 支持.拟议的实验和培训活动将使候选人能够发表顶级 眼科研究工作，并独特地定位他作为一个独立的主要研究者追求新的 用于视网膜疾病如青光眼的治疗剂。 青光眼是美国第二大致盲原因，至少有3，000，000人受到影响。 到2030年，如果没有新的治疗方法，这个数字可能会增加60%。由于无法 RGC死亡导致的视力丧失是不可逆的， 会导致永久性失明我们的初步实验使用基于CRISPR/Cas9的体内正向表达， 基因筛选发现，敲低CCCTC结合因子（CTCF）可以强烈促进 RGC的轴突再生，而不增加整体RGC的存活， 特异性识别蛋白1（SSRP 1）显著保护RGC存活而不促进轴突再生。 为了探索调节RGC存活和再生的潜在表观遗传机制， 本研究的主要目的是：（1）利用ATAC技术研究损伤诱导的RGC染色质重塑 测序（指导阶段）;（2）评估介导CTCF和 SSRP 1在损伤后（指导和独立阶段）;和（3）评估 表观遗传调节剂在青光眼模型（独立阶段）。拟议研究的结果将提供 深入和定量的见解，为什么以及如何RGC的再生命运是预先确定的， 表观基因组的观点，这可以直接转化为新的治疗视神经病变。