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

基本信息

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
项目类别：
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

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

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项目摘要

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.

项目摘要面部形态发生涉及面部原始的定向生长，直到它们与一个其他。目前，原始的产物被认为主要由散装组织驱动由间充质组织中的增殖梯度引起。但是，我们的实验室和其他人发现其他机制（例如定向的细胞迁移和上皮引导生长）也可能有助于。在外胚层产生的肢体芽，拉伸力已被证明是指导细胞方向和生长，肢体芽的增殖梯度在不掺入的情况下无法准确模拟形态发生定向细胞行为。我们的实验室以前已经表明，面部的间充质细胞是极化的，并且成纤维细胞生长因子（FGF）的激活信号传导破坏了这种极性，同时导致异常生长原始。细胞周围的细胞外基质（ECM）提供了所需的结构支架通过组织产生和引导力，上皮中的ECM可以在区域指导或限制生长。我假设ECM通过组成的区域差异和遵守。我将以两个主要目的检验这一假设。在第一个目标中，我将研究FGF信号如何影响面部原基中的ECM物理特性和细胞组织。我将定量映射 ECM蛋白质和ECM依从性的组成与共聚焦显微镜和原子力的依从性显微镜，然后将这些度量的区域差异与面部形状变化相关联。这个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蜂窝组织。在第二个目标中，我将通过改变直接增加或降低组织合规性胶原蛋白交联，然后重复与第一个目标相同的分析。第二个目标将确定ECM是否刚度直接引导生长，而间充气在其周围被动地符合或有反馈时在ECM和主动重塑环境以引导生长的细胞之间。面部原则的失败 appose和Fuse会导致面部裂纹。了解ECM在定向增长中的作用对于了解面部出生缺陷的病因。