Determining the Role of Extracellular Matrix Compliance and Composition on Facial Morphogenesis

确定细胞外基质顺应性和成分对面部形态发生的作用

• 批准号: 10306333
• 负责人: Nicholas Jean Hanne
• 金额: $ 7.06万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-01 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10306333
• 关键词: 3-Dimensional; Actomyosin; Affect; Area; Atomic Force Microscopy; Basement membrane; Cell Polarity; Cell Proliferation; Cell Shape; Cells; Cleft Palate; Cleft lip with or without cleft palate; Collagen; Confocal Microscopy; Congenital Abnormality; Craniofacial Abnormalities; Cytoskeleton; DNA Sequence Alteration; Data; Development; Dysmorphology; Ectoderm; Embryo; Environment; Enzymes; Epithelial; Etiology; Exposure to; Extracellular Matrix; Extracellular Matrix Proteins; Face; Failure; Family; Feedback; Genes; Genetic; Goals; Growth; Growth Factor; Human; Knock-out; Laminin; Lathyrus; Lead; Limb Bud; Maps; Measures; Medial; Mesenchymal; Mesenchyme; Modeling; Molecular; Morphogenesis; Morphology; Movement; Mus; Mutation; Nose; Outcome Measure; Parents; Patients; Play; Premaxillary palate; Primordium; Process; Process Measure; Property; Protein-Lysine 6-Oxidase; Publishing; Research; Role; S-Phase Fraction; Shapes; Signal Transduction; Spectroscopy, Fourier Transform Infrared; Structure; Surface; Testing; Thick; Tissues; Transmission Electron Microscopy; Work; Zebrafish; cell behavior; cell growth; cell motility; craniofacial; crosslink; interest; member; physical property; prevent; regional difference; response; scaffold; small molecule

PROJECT SUMMARY Facial morphogenesis involves directed outgrowth of the facial primordia until they appose and fuse with one another. Currently, outgrowth of primordia is considered to be driven primarily by bulk tissue displacement caused by proliferation gradients in the mesenchymal tissue. However, our lab and others have found that other mechanisms, such as directed cellular migration and epithelium guided growth, likely contribute as well. In the limb bud, tensile forces generated by the ectoderm have been shown to direct cellular orientation and growth, and proliferation gradients across the limb bud cannot accurately model morphogenesis without incorporating directed cellular behaviors. Our lab has previously shown that mesenchymal cells in the face are polarized and that activation of fibroblastic growth factor (FGF) signaling disrupts this polarity while causing aberrant outgrowth of the primordium. The extracellular matrix (ECM) around cells provides the structural scaffolding required to generate and guide force through tissue, and the ECM in the epithelium may guide or constrain growth regionally. I hypothesize that the ECM directs morphogenesis in the face via regional differences in composition and compliance. I will test this hypothesis with two main aims. In the first aim I will examine how FGF signaling affects ECM physical properties and cellular organization in the facial primordium. I will quantitatively map the composition of ECM proteins and ECM compliance across the face with confocal microscopy and atomic force microscopy, then relate regional differences in these measures to facial shape change. This aim will elucidate mechanisms by which molecular signaling of fibroblastic growth factor controls morphogenesis through the ECM and cellular organization. In the second aim, I will increase or decrease tissue compliance directly by altering collagen crosslinking, then repeat the same analyses as the first aim. The second aim will determine if ECM stiffness directs outgrowth directly while the mesenchyme passively conforms around it, or if there is feedback between ECM and cells that actively remodel the environment to direct growth. Failure of the facial primordium to appose and fuse leads to facial clefting. Understanding the role of ECM in directional growth is important for understanding the etiology of birth defects in the face.

项目概要 面部形态发生涉及面部原基的定向生长，直到它们与一个原基并列并融合 其他。目前，原基的生长被认为主要是由大量组织移位驱动的 由间充质组织的增殖梯度引起。然而，我们的实验室和其他人发现，其他 定向细胞迁移和上皮引导生长等机制也可能有所贡献。在 肢芽，外胚层产生的拉力已被证明可以指导细胞定向和生长， 如果不合并，整个肢芽的增殖梯度就无法准确地模拟形态发生 定向细胞行为。我们的实验室之前已经表明，面部的间充质细胞是极化的，并且 成纤维细胞生长因子 (FGF) 信号的激活会破坏这种极性，同时导致异常生长 原基的。细胞周围的细胞外基质 (ECM) 提供了所需的结构支架 产生并引导力穿过组织，上皮中的 ECM 可以引导或限制区域生长。 我假设 ECM 通过成分和结构的区域差异来指导面部的形态发生。 遵守。我将通过两个主要目标来检验这个假设。第一个目标是研究 FGF 信号如何影响 面部原基中的 ECM 物理特性和细胞组织。我将定量绘制 通过共焦显微镜和原子力观察 ECM 蛋白的组成以及整个面部的 ECM 顺应性 显微镜，然后将这些措施的区域差异与面部形状的变化联系起来。这一目标将阐明 成纤维细胞生长因子的分子信号传导通过 ECM 控制形态发生的机制 和细胞组织。在第二个目标中，我将通过改变直接增加或减少组织顺应性 胶原交联，然后重复与第一个目标相同的分析。第二个目标将决定 ECM 是否 僵硬直接引导生长，而间充质被动地顺应其周围，或者如果有反馈 细胞外基质和细胞之间的相互作用，主动重塑环境以直接生长。面部原基衰竭 并置和融合会导致面部裂开。了解 ECM 在定向生长中的作用对于 了解面部出生缺陷的病因。