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.

项目摘要/摘要 拟议的研究是一项为期五年的职业发展研究计划，重点是剖析表观遗传 视网膜神经节细胞（RGC）的变性和复兴的调节小鼠模型 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 在表观基因组分析技术中，视神经病变的机制和神经创力的发展 通过在波士顿儿童医院和哈佛医学的Zhigang He博士的心理团队中整合疗法 学校，哈佛大学的约书亚·萨内斯博士和哈佛脑科学倡议，杰弗里·戈德伯格博士 斯坦福大学和拜尔斯眼科研究所，以及哈佛干细胞的杰森·布恩罗斯特罗博士和再生 生物学（SCRB）部门和麻省理工学院和哈佛大学广泛研究所。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，是他的特定科学和技术顾问 支持。拟议的实验和培训活动将使候选人能够发布顶级 眼科研究的工作和独特地将他定位为独立的首席研究员，追求小说 残留疾病的治疗剂，例如青光眼。 青光眼是美国失明的第二大主要原因，至少有300万人受到影响。 如果没有新的疗法，到2030年，这个数字可能会增加60％。由于不稳定 中枢神经系统受伤后重生，RGC死亡导致的视力丧失是不可逆的，并且 将导致永久失明。我们使用基于CRISPR/CAS9的体内前进的初步实验 遗传筛查发现损伤诱导的表观遗传调节剂，例如CCCTC- 结合因子（CTCF）可以稳健地促进RGC轴突再生或/和/和/and的生存。探索基础 拟议的研究将专门追求调查RGC生存和再生的表观遗传机制 以下目的：（1）通过ATAC测序（指导）在RGC中对损伤诱导的染色质进行了重塑 阶段）; （2）评估生存和再生调节剂媒体差异效应的机制 受伤（指导和独立阶段）； （3）评估表观遗传学的机制和影响 青光眼模型中的调节剂（独立阶段）。在过去的两年中，在这个K99奖中，我有 成功完成了AIM 1和AIM2中的所有实验和目标，并发表了两个手稿 神经元于2022年（被选为封面故事）。在我的K奖的延长期间，我将进一步购买 我在青光眼相关小鼠模型中的方法的翻译研究。拟议研究的结果 将对RGC的再生命运的原因和定量见解提供深入和定量的见解。 从表观基因组的角度确定，可以将其直接转换为新疗法的光学疗法 神经病。