Role of retinoic acid signaling in fovea development

视黄酸信号在中央凹发育中的作用

基本信息

批准号：
10541865
负责人：
Susana Isabel M M A da Silva
金额：
$ 39.53万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-01-01 至 2026-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10541865
关键词：
3-Dimensional Address Affect Age related macular degeneration Area Biological Models Birds Blindness Cell Therapy Cell Transplantation Chick Cone Data Development Disease Disease model Doxycycline Drug Screening Endowment Enzymes Experimental Models Face Gene Transfer Techniques Genetic Goals High Prevalence Human In Vitro Individual Macular degeneration Mammals Microscopy Molecular Organ Organoids Pathway interactions Pattern Primates Property Protocols documentation Reading Regulation Reporter Retina Retinal Diseases Rod Rodent Role Signal Transduction Source Speed Structure System Testing Transgenic Organisms Translating Tretinoin United States Vision Visual Acuity burden of illness clinically relevant density effective therapy fetal fovea centralis ganglion cell high resolution imaging high-throughput drug screening human stem cells in vitro Model in vivo induced pluripotent stem cell innovation insight macula model organism neural circuit novel pharmacologic programs retinogenesis societal costs spatiotemporal therapy development tool translational potential

项目摘要

PROJECT SUMMARY: The fovea is a highly specialized retinal structure located within the macula responsible for high- acuity vision, which in humans enables critical tasks such as reading and recognizing faces. Foveal diseases such as macular degeneration are highly debilitating and often lead to blindness. However, there are no current available treatments or therapies for fovea-specific disorders. A major reason underlying this limitation is the lack of suitable experimental systems for foveal studies, as only a small number of species have fovea-like high acuity areas (HAA). Indeed, among mammals, only a subset of primates has foveated retinas. However, avian species including the chick, possess fovea-like HAA and can be used as a powerful model system to study the molecular underpinnings of fovea formation in vivo. Human stem cell-derived retinal organoids (hRetOrg) have emerged as a promising in vitro system to study human retinal disorders but current protocols fail to generate foveae. We have recently found that retinoic acid (RA) signaling is highly patterned during foveogenesis in humans and is required for HAA formation in the chick. Our hypothesis is that recapitulation of endogenous patterns of RA signaling in hRetOrg is sufficient to induce fovea formation. To test this hypothesis, we will utilize the chick system to elucidate the spatiotemporal properties of RA-dependent fovea induction in vivo (Aim 1); longitudinally characterize the dynamics of RA signaling and expression profile of RA enzymes during retinogenesis period in hRetOrg (Aim 2); experimentally manipulate RA signaling to recapitulate the spatiotemporal patterns of RA found in vivo in the in vitro hRetOrg system (Aim 3). To achieve this we have developed an innovative molecular strategy based on transposon- and doxycycline-inducible systems for precise spatial and temporal regulation of transgenesis and optimized high-speed high-resolution imaging analysis of whole chick retinas and 3D hRetOrg. This effort will not only advance our current understanding of the mechanistic role of RA signaling as a fovea regulator but also generate an innovative and much needed foveated hRetOrg system with high translational potential for diseases affecting the fovea through disease modeling, high-throughput drug screening and as a source of clinically-relevant fovea-specific cells for transplantation.

项目概要：中央凹是位于黄斑内的高度专业化的视网膜结构，负责高敏锐的视觉，使人类能够完成阅读和识别面孔等关键任务。黄斑变性等黄斑变性等黄斑中心凹疾病会导致患者高度衰弱，并常常导致失明。然而，目前没有针对中央凹特异性疾病的可用治疗方法或治疗方法。一个这种限制的主要原因是缺乏合适的中央凹实验系统研究表明，只有少数物种具有中央凹样高敏锐度区域（HAA）。的确，在哺乳动物中，只有一小部分灵长类动物具有中心凹视网膜。然而，鸟类物种包括雏鸡在内，都具有类似中央凹的HAA，可以作为强大的模型系统进行研究体内中央凹形成的分子基础。人类干细胞来源的视网膜类器官 (hRetOrg) 已成为研究人类视网膜疾病的有前景的体外系统，但当前的协议无法生成中央凹。我们最近发现视黄酸（RA）信号传导在人类视网膜中央凹形成过程中，它是高度模式化的，并且是雏鸡 HAA 形成所必需的。我们的假设是 hRetOrg 中 RA 信号传导的内源模式的重演是足以诱导中央凹形成。为了检验这个假设，我们将利用小鸡系统来阐明体内 RA 依赖性中央凹诱导的时空特性（目标 1）；纵向表征 RA 信号传导的动态和 RA 酶的表达谱 hRetOrg 视网膜发生期间（目标 2）；实验性地操纵 RA 信号概括在体外 hRetOrg 系统中体内发现的 RA 时空模式（目的 3）。为了实现这一目标，我们开发了一种基于转座子的创新分子策略和多西环素诱导系统，用于转基因的精确空间和时间调节并对整鸡视网膜和 3D 进行优化的高速高分辨率成像分析 hRetOrg。这项工作不仅将增进我们目前对 RA 机制作用的理解作为中央凹调节器的信号传导，但也产生创新且急需的中央凹 hRetOrg 系统对于通过疾病影响中央凹的疾病具有高转化潜力建模、高通量药物筛选以及作为临床相关的中央凹特异性的来源用于移植的细胞。