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: 10845925
• 负责人: Feng Tian
• 金额: $ 2.84万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 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 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10845925
• 关键词: Epigenomic; mechanisms; regulating; RGC; survival

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 injury induced epigenetic regulators, such as CCCTC- binding factor (CTCF), can robustly promote RGC axonal regeneration or/and survival. 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 survival and regeneration regulators upon injury (mentored and independent phase); and (3) To assess the mechanisms and effects of epigenetic regulators in a glaucoma model (independent phase). For the past two years during this K99 award, I have successfully accomplished all experiments and goals in Aim 1 and Aim2, with two manuscripts published on Neuron in 2022 (and selected as cover story). For the extended period of my K award, I will further pursue the translational study of my approaches in glaucoma relevant mouse models. 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.

项目概要/摘要 拟议的研究是一项为期五年的职业发展研究计划，重点剖析表观遗传 视神经挤压小鼠模型视网膜神经节细胞（RGC）变性和再生的调节 和青光眼。该候选人目前是 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 表观基因组分析技术、视神经病变机制和神经再生发展 整合波士顿儿童医院何志刚博士和哈佛医学院的导师团队进行治疗 哈佛大学的 Joshua Sanes 博士和哈佛大学脑科学计划的 Jeffery Goldberg 博士 斯坦福大学和拜尔斯眼科研究所以及哈佛大学干细胞和再生中心的 Jason Buenrostro 博士 麻省理工学院和哈佛大学的生物学（SCRB）系和博德研究所。该候选人还招募了丹尼尔博士 Geschwind 是他当前和拟议研究的合作者，作为他特定科学和技术的顾问 支持。拟议的实验和培训活动将使候选人能够发表顶级论文 眼科研究工作并将他独特地定位为追求新颖的独立首席研究员 青光眼等视网膜疾病的治疗方法。 青光眼是美国第二大失明原因，至少有 3,000,000 人受到影响。 如果不能开发出新的疗法，到 2030 年这个数字可能会增加 60%。由于无能力 中枢神经系统损伤后无法再生，RGC死亡导致的视力丧失是不可逆的 将导致永久性失明。我们使用基于 CRISPR/Cas9 的体内前向进行初步实验 基因筛选发现损伤诱导的表观遗传调节因子的降低，例如 CCCTC- 结合因子（CTCF），可以强有力地促进 RGC 轴突再生或/和存活。探索底层 调节 RGC 存活和再生的表观遗传机制，拟议的研究将专门追求 目标如下：（1）通过 ATAC 测序（指导）来分析 RGC 中损伤诱导的染色质重塑 阶段）; (2) 评估介导生存和再生调节剂差异作用的机制 受伤时（指导和独立阶段）； (3) 评估表观遗传的机制和影响 青光眼模型中的调节器（独立阶段）。在过去两年的 K99 颁奖期间，我 成功完成Aim 1和Aim2的所有实验和目标，两篇手稿发表在 2022 年《神经元》（并被选为封面故事）。在我的 K 奖延长期间，我将进一步追求 我的方法在青光眼相关小鼠模型中的转化研究。拟议研究的结果 将为 RGC 的再生命运为何以及如何预提供深入和定量的见解 从表观基因组角度确定，可直接转化为视神经疾病的新疗法 神经病。