Role of retinoic acid signaling in fovea development

视黄酸信号在中央凹发育中的作用

• 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
• 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

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.

项目摘要： Fovea是一个高度专业化的视网膜结构 敏锐的愿景，在人类中可以实现关键任务，例如阅读和识别面孔。 诸如黄斑变性之类的凹起疾病高度令人衰弱，并且常常导致失明。 但是，目前尚无类动植物特异性疾病的可用疗法或疗法。一个 该限制的主要原因是缺乏合适的Foveal实验系统 研究，因为只有少数物种具有中央凹形的高敏锐区域（HAA）。的确， 在哺乳动物中，只有一部分灵长类动物具有凹陷的视网膜。但是，鸟类 包括小鸡，具有类似Fovea的HAA，可以用作研究的强大模型系统 体内中央凹形形成的分子基础。人类干细胞衍生的视网膜器官 （Hretorg）已成为研究人类视网膜疾病的有前途的体外系统，但 当前的协议无法生成Foveae。我们最近发现视黄酸（RA）信号传导 在人类的卵生期间是高度图案的，是雏鸡中HAA形成所必需的。 我们的假设是，Hretorg中RA信号传导的内源性模式的概括为 足以诱导中央凹的形成。为了检验这一假设，我们将利用小鸡系统 阐明体内RA依赖性中央凹诱导的时空特性（AIM 1）； 纵向表征Ra酶的RA信号传导和表达曲线的动力学 在Hretorg的视网膜生成期间（AIM 2）；实验操纵RA信号传导 概述体内在体外Hretorg系统中发现的RA的时空模式（AIM 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 和多西环素诱导的系统，用于转基因的精确空间和时间调节 并优化了整个小鸡视网膜和3D的高速高分辨率成像分析 Hretorg。这项努力不仅会提高我们当前对RA机械作用的理解 作为中央凹调节器的信号传导，但也产生了创新且急需的FOVEATEN Hretorg系统具有很高的转化潜力，可影响中央凹的疾病 建模，高通量药物筛查以及作为临床上的中央凹特异性的来源 用于移植的细胞。