Stepwise Coordination of Eye Morphogenesis by Extracellular Matrix

细胞外基质对眼睛形态发生的逐步协调

• 批准号: 10583547
• 负责人: Kristen M Kwan
• 金额: $ 38.13万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10583547
• 关键词: Automobile Driving; Basement membrane; Biological Models; Bruch' s basal membrane structure; Cell Death; Cell Size; Cell Survival; Cell Transplantation; Cell physiology; Cells; Chondroitin Sulfate A; Chondroitin Sulfate Proteoglycan; Collaborations; Coloboma; Complex; Computational Technique; Computer Analysis; Computing Methodologies; Data; Defect; Deposition; Development; Ectoderm; Embryo; Environment; Epithelium; Event; Exposure to; Extracellular Matrix; Eye; Eye Development; Focal Adhesions; Four-dimensional; Fresh Water; Gene Expression Profiling; Glycoproteins; Goals; Human; Image; Imaging Techniques; Impairment; In Vitro; Laminin; Methods; Microscopy; Modeling; Molecular; Molecular Genetics; Morphogenesis; Morphology; Movement; Mutation; Neural Crest; Neural Crest Cell; Neural Retina; Newborn Infant; Nidogen; Ocular Pathology; Optic vesicle; Optics; Organoids; Pathway interactions; Physiological; Positioning Attribute; Process; Regenerative Medicine; Retina; Role; Series; Shapes; Signal Transduction; Structural defect; Structure; Structure of retinal pigment epithelium; Tenascin; Testing; Tissue Engineering; Tissue Model; Tissues; Transplantation; Visual impairment; Visualization; Work; Zebrafish; cell behavior; cell motility; cell type; experimental study; extracellular; eye formation; imaging modality; in vivo; innovation; lens; membrane assembly; model organism; molecular dynamics; optic cup; progenitor; programs; protein crosslink; response; single cell sequencing; spatiotemporal; teleost fish; timeline; versican; zebrafish development

Project Summary Developmental defects in eye structure commonly account for visual impairment in newborns. Proper eye structure is initially established via the process of optic cup morphogenesis, during which a series of complex cell and tissue rearrangements transforms the optic vesicle into the optic cup, with neural retina and retinal pigmented epithelium (RPE) enwrapping the newly formed lens. With advances in imaging and computational analysis, work from our lab and others has begun to reveal the cellular events underlying optic cup morphogenesis, however, molecular control of these processes still remains poorly understood. A compelling candidate to play a role in controlling optic cup morphogenesis is the extracellular matrix (ECM), a complex, glycoprotein-rich layer that regulates cell survival, movement, signaling, and polarity. Mutations in certain ECM components can lead to ocular pathologies, such as coloboma, suggesting specific requirements during optic cup morphogenesis. Understanding extrinsic control of morphogenesis also has implications for organoid approaches and strategies. Our previous data indicate that a core molecule, laminin, elicits diverse cellular responses in different eye regions. We also found that separate eye domains are exposed to distinct ECM microenvironments, some of which are assembled via tissue-tissue interactions: specifically, neural crest is required to build basement membrane around the RPE. These data suggest that unique ECM microenvironments may be a crucial driver of regional eye morphogenetic events. Zebrafish provide an ideal model system to study this process: optical transparency and rapid development offer a unique opportunity to directly observe and molecularly dissect eye formation in vivo. We previously developed 4-dimensional imaging and computational techniques to track and visualize cell movements throughout optic cup morphogenesis, and recently, methods for automated quantitative analysis of retinal cell size, shape and orientation. This puts us in a unique position to analyze specific morphogenetic defects arising when particular matrix components are disrupted. In this proposal, we will dissect the region-specific roles of ECM factors, including nidogens, tenascin-C, mmp2, and versican, during eye morphogenesis. We hypothesize that dynamic, region-specific ECM microenvironments elicit unique developmental and morphogenetic responses from distinct eye progenitor domains to drive optic cup morphogenesis. Combining molecular genetics with innovative 4-dimensional live imaging and computational methods, we will test this hypothesis in the following specific aims: (1) determine how ECM microenvironment controls retina morphogenesis and organization; (2) determine how ECM modulatory factors control RPE morphogenesis; and (3) determine functional requirements for tissue contributions to specific ECM microenvironments. The experiments proposed will define the spatiotemporal dynamics of ECM activity and distinct cellular functions executed by region-specific ECM factors during crucial steps of eye formation.

项目摘要 眼睛结构的发育缺陷通常是新生儿视力障碍的原因。固有眼 结构最初是通过视杯形态发生的过程建立的，在此过程中， 细胞和组织重排将视泡转化为视杯， 包裹新形成的透镜的色素上皮（RPE）。 随着成像和计算分析的进步，我们实验室和其他人的工作已经开始揭示 然而，视杯形态发生背后的细胞事件，这些过程的分子控制仍然存在 不太了解。在控制视杯形态发生中起作用的一个令人信服的候选者是 细胞外基质（ECM），一种复杂的富含糖蛋白的层，其调节细胞存活、运动、信号传导， 和极性。某些ECM组分中的突变可导致眼部病变，例如缺损， 提示视杯形态发生过程中的特殊要求。理解外部控制 形态发生也对类器官方法和策略有影响。我们以前的数据表明， 核心分子，层粘连蛋白，在不同的眼部区域引起不同的细胞反应。我们还发现， 眼域暴露于不同的ECM微环境，其中一些是通过组织-组织组装的 相互作用：具体而言，需要神经嵴在RPE周围建立基底膜。这些数据 这表明独特的ECM微环境可能是区域性眼形态发生事件的关键驱动因素。 斑马鱼为研究这一过程提供了理想的模式系统：光学透明性和快速发育 提供了一个独特的机会，直接观察和分子解剖眼睛的形成在体内。我们之前 开发了4维成像和计算技术来跟踪和可视化细胞运动 以及最近用于视网膜细胞的自动定量分析的方法 大小、形状和方向。这使我们处于一个独特的位置，以分析特定的形态发生缺陷， 当特定的基质成分被破坏时。在本提案中，我们将分析以下各区域的具体作用： ECM因子，包括巢蛋白、腱生蛋白-C、mmp 2和多功能蛋白聚糖，在眼形态发生中。 我们假设，动态的，区域特异性ECM微环境引起独特的发育和 来自不同眼祖域的形态发生反应来驱动视杯形态发生。结合 分子遗传学与创新的四维实时成像和计算方法，我们将测试这一点， 本研究的主要目的是：（1）确定ECM微环境如何控制视网膜 （2）确定ECM调节因子如何控制RPE形态发生;和 (3)确定组织对特定ECM微环境的贡献的功能要求。的 提出的实验将定义ECM活性的时空动态和不同的细胞功能 在眼睛形成的关键步骤中由区域特异性ECM因子执行。