Comparative Single Cell Profiling of Retinal Development

项目摘要 老年性黄斑变性(AMD)影响视网膜的中央区域，该区域对高视力非常重要 日间视力，导致失明或严重视力受损，原因是 以锥体为主的中心凹。视网膜前体细胞定向分化为视锥感光细胞显示 有望成为AMD和其他视网膜营养不良的治疗方法。然而，常用的模式生物，如 因为老鼠的视网膜以杆状为主，这使得它们不适合作为人类中心凹的模型。13行 然而，磨砂松鼠视网膜的视锥/视杆比和视锥亚型组成非常匹配。 那是人类的中心凹。该项目将使用13行地松鼠作为模型来鉴定基因 驱动视网膜锥体发育的调控网络，并测试候选人的功能是否获得 基因可以促进小鼠的锥体发育。我假设基因调控的差异激活 控制光感受器发育的网络是视锥/视杆比率差异的基础 小鼠和13条线的地松鼠以及在人类外周视网膜和视锥之间- 优势中心凹。为了解决这一假设，我提出了两个目标。目标1：全面剖析视网膜 利用scRNA-seq和scATAC-seq在单细胞分辨率下对13线地松鼠的研究 确定控制视锥/视杆感光细胞进化变化的候选基因调控网络 比率这些数据将使我能够确定控制13-LINE发展的关键基因调控网络 磨碎的松鼠视网膜，并将找出光感受器产生差异的可能原因 13只地松鼠和小鼠之间的亚型，以及中心凹和周围人类视网膜之间的亚型。 目的2：测试候选基因在小鼠体内过度表达的功能后果 在13-LGS中推广圆锥体规格。这将确定控制锥体/杆需要进行哪些更改 在发育中的小鼠视网膜中的比率。通过了解控制光感受器的基因调控网络 在视网膜中的比例，我将能够指导小鼠视网膜祖细胞产生视锥主导的视网膜， 最终为AMD的进展和细胞治疗提供了一个改进的动物模型。

Project Summary Age-related macular degeneration (AMD) affects the central region of the retina important for high-acuity daytime vision, causing blindness or severe vision impairment as a result of the loss of photoreceptors from the cone-dominant fovea. Directed differentiation of retinal progenitor cells into cone photoreceptors shows promise as a therapy for AMD and other retinal dystrophies. However, commonly used model organisms such as the mouse have rod-dominant retinas, which makes them poor models for the human fovea. 13-lined ground squirrel retinas, however, have a cone/rod ratio and cone subtype composition that closely matches that of the human fovea. This project will use the 13-lined ground squirrel as a model to identify gene regulatory networks that drive development of retinal cones and test whether gain of function of candidate genes can promote cone development in mice. I hypothesize that differential activation of gene regulatory networks controlling photoreceptor development underlie the differences in the cone/rod ratio between the mouse and 13-lined ground squirrel and between the peripheral human retina and the cone- dominant fovea. To address this hypothesis, I propose two Aims. Aim 1: Comprehensively profile retinal development in the 13-lined ground squirrel at single-cell resolution using both scRNA-seq and scATAC-seq to identify candidate gene regulatory networks that control evolutionary changes in the cone/rod photoreceptor ratio. This data will allow me to identify key gene regulatory networks that control development of the 13-lined ground squirrel retina and will identify plausible causes for the difference in the production of photoreceptor subtypes between 13-lined ground squirrel and mouse and between the fovea and peripheral human retina. Aim 2: Test the functional consequences of overexpression of candidate genes in mice that are predicted to promote cone specification in 13-LGS. This will identify which changes are necessary to control the cone/rod ratio in the developing mouse retina. By understanding the gene regulatory networks that control photoreceptor proportions in the retina, I will be able to direct mouse retinal progenitors to generate a cone-dominant retina, ultimately providing an improved animal model for AMD progression and cell therapy.