Photoreceptor Regeneration in a Murine Model of Leber Congenital Amaurosis

项目名称 Leber先天性巨结肠小鼠模型的光感受器再生 摘要 Leber先天性黑色素瘤(LCA)是一种起病早、严重的遗传性视网膜变性，可导致 儿童期失明。基因治疗可有效恢复Lca5gt/GT小鼠模型的视功能 在出现严重的视网膜变性之前，如果给新生的小鼠注射这种病毒，就会感染疾病。但是，在 中晚期疾病发生后感光细胞死亡，基因治疗不再 有效。基于细胞的疗法为晚期视网膜再生光感受器带来了巨大希望 退化，但进展受到整合效率低和复杂细胞的限制 与现有视网膜细胞的相互作用。 光感受器前体代表了不同类型的细胞的异质性池，这些细胞具有潜在的 在发育和患病的视网膜中分化为成熟的感光细胞。使用无偏的单电池 RNA转录组学，我鉴定了几个新的CRX光受体前体群体，包括 CRX/Nfix和CRX/Slc39a1细胞。这项提议的科学目标是再生 Lca5gt/GT小鼠视网膜变性模型中的光感受器 光感受器前体的群体和内源性修复通路的激活在Müler神经胶质细胞中起重要作用。 该提案的目的是1)将CRX/NFIX和CRX/SLc39a1祖细胞移植到视网膜下 Lca5gt/GT小鼠，以及2)通过基因治疗激活Lca5gt/GT小鼠内源性Müler神经胶质修复通路。 除了这些科学贡献外，这项提案还概述了一项结构化的、重点突出的培训计划，该计划将 让我掌握技能和专业知识，这些技能和专业知识将成为我开发基于细胞技术的职业生涯的基础 视网膜疾病的治疗方法。宾夕法尼亚大学眼科是一个理想的选择 为培训内科科学家进行眼科研究提供环境，并将提供保护时间， 成功过渡到独立所需的资源和指导。

PROJECT TITLE Photoreceptor Regeneration in a Murine Model of Leber Congenital Amaurosis ABSTRACT Leber congenital amaurosis (LCA) is an early-onset, severe inherited retinal degeneration that results in childhood blindness. Gene therapy can effectively restore visual function in the Lca5gt/gt mouse model of the disease when delivered to neonatal mice before significant retinal degeneration is present. However, in moderate- and late- stage disease after photoreceptor cell death has occurred, gene therapy is no longer effective. Cell-based therapies hold great promise for regenerating photoreceptors in late-stage retinal degenerations, but progress has been limited by low efficiencies of integration and complex cellular interactions with existing retinal cells. Photoreceptor precursors represent a heterogeneous pool of diverse cells types that have the potential to differentiate into mature photoreceptor cells in the developing and diseased retina. Using unbiased single-cell RNA transcriptomics, I identified several novel populations of Crx+ photoreceptor precursors including Crx+/Nfix+ and Crx+/Slc39a1+ cells. The scientific objectives of this proposal are to regenerate photoreceptors in the Lca5gt/gt mouse model of retinal degeneration, by the transplantation of these novel populations of photoreceptor precursors and by the activation of endogenous repair pathways in Müller glia. The Aims of the proposal are 1) Subretinal transplantation of Crx+/Nfix+ and Crx+/Slc39a1+ progenitor cells in Lca5gt/gt mice, and 2) Activation of endogenous Müller glial repair pathways by gene therapy in Lca5gt/gt mice. In addition to these scientific contributions, this proposal outlines a structured, focused training plan that will equip me with the skills and expertise that will serve as the foundation for my career in developing cell-based therapies for retinal disease. The Department of Ophthalmology at the University of Pennsylvania is an ideal environment for training physician scientists in ophthalmic research, and will provide the protected time, resources, and mentorship needed for a successful transition to independence.