Photoreceptor Regeneration in a Murine Model of Leber Congenital Amaurosis

项目名称 Leber先天性黑蒙小鼠模型的光感受器再生 摘要 Leber先天性黑蒙（LCA）是一种早发性、严重的遗传性视网膜变性， 儿童失明。基因治疗可以有效地恢复Lca 5gt/gt小鼠模型的视觉功能， 当在显著的视网膜变性存在之前递送给新生小鼠时，但在 光感受器细胞死亡后的中晚期疾病， 有效基于细胞的疗法在晚期视网膜病变中再生光感受器方面前景广阔 退化，但进展受到整合效率低和复杂的细胞 与现有视网膜细胞的相互作用。 光感受器前体代表了不同细胞类型的异质池，其具有潜在的 在发育和患病的视网膜中分化成成熟的感光细胞。使用无偏单细胞 RNA转录组学，我确定了几个新的Crx+感光细胞前体群体， Crx+/Nfix+和Crx+/Slc 39 a1+细胞。这项提案的科学目标是再生 视网膜变性的Lca 5gt/gt小鼠模型中的光感受器，通过移植这些新的 群体的感光细胞前体和Müller胶质细胞的内源性修复途径的激活。 该提案的目的是：1）视网膜下移植Crx+/Nfix+和Crx+/Slc 39 a1+祖细胞， Lca 5gt/gt小鼠，和2）在Lca 5gt/gt小鼠中通过基因治疗激活内源性Müller神经胶质修复途径。 除了这些科学贡献外，该提案还概述了一个结构化的、有重点的培训计划， 使我具备技能和专业知识，这将成为我在开发基于细胞的 视网膜疾病的治疗。宾夕法尼亚大学的眼科是一个理想的 培训眼科研究医生科学家的环境，并将提供受保护的时间， 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.