Defining an Integrated Signaling Network That Patterns the Craniofacial Skeleton

定义一个模拟颅面骨骼的集成信号网络

基本信息

批准号：
8750599
负责人：
David E. Clouthier
金额：
$ 67.12万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-07-01 至 2019-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8750599
关键词：
Address Affect Alagille Syndrome Automobile Driving Bone Morphogenetic Proteins Branchial arch structure Buffers Candidate Disease Gene Cleft Palate Collection Competence Complex Computer Analysis Computer Simulation Congenital Abnormality Data Databases Defect Development Disease Dorsal Embryo Endothelin-1 Enhancers Ensure Etiology Event Face Feedback Fishes Gene Expression Gene Mutation Gene Targeting Genes Genetic Genetic Enhancer Element Goals High-Throughput RNA Sequencing Human Interdisciplinary Study Jaw Mediating Micrognathism Mining Modeling Molecular Profiling Morphogenesis Mus Mutate Noise Notch Signaling Pathway Outcome Pathway interactions Pattern Phenotype Play Positioning Attribute Regulator Genes Regulatory Element Resistance Role Shapes Signal Pathway Signal Transduction Silicon Skeletal Development Skeleton Structural Congenital Anomalies Syndrome System Testing Time To specify Transgenic Mice Transgenic Organisms Work Zebrafish base clinically relevant craniofacial gene function genetic analysis improved in vivo in vivo imaging malformation mathematical model mouse model mutant notch protein novel progenitor programs public health relevance response simulation skeletal spatiotemporal three dimensional structure tool transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): Much of the craniofacial skeleton arises from the pharyngeal arches, 3D structures that undergo complex changes in shape and gene expression over time. However, detailed analyses of early gene expression profiles in the arches have not been performed in vivo. Without such information, it is impossible to predict how gene expression changes will affect subsequent skeletal development. Our labs have extensively studied the endothelin1 (Edn1), bone morphogenetic protein (Bmp), Wnt and Jag/Notch signaling pathways in the arches, as these four pathways pattern the dorsal-ventral (D-V) axis of the facial skeleton. We have shown that Edn1 and Bmp signaling initially promote ventrally- expressed genes and later subdivide the arches into separate D-V sub-domains. Our preliminary data suggest that Wnt signaling controls competence to respond to Edn1/Bmp and that these three pathways are opposed by Jag/Notch signaling. The pathways regulated by these four signals are highly dynamic, containing multiple feedback loops and crosstalk that create a robust system resistant to perturbation. With a collection of mouse and zebrafish mutants in all four signals, we are in a unique position to assess conservation of gene expression across species in sufficient detail for computational modeling. Our goal is to use these models to predict in silicon facial defects observed following genetic perturbations of these signals. Our dual-species approach will identify new candidate genes and generate models that are clinically relevant to human craniofacial genetics. To address these goals we will pursue two specific aims. In Aim 1, we hypothesize that while Edn1, Bmp, Wnt and Jag/Notch signaling are all critical for establishing the initial identities of skeletal progenitor in the arches along the D-V axis, they each play distinct roles. We will address this hypothesis by using high-throughput RNA sequencing to define early changes in gene expression in mutants of all four pathways. These Early Response Profiles (ERPs) will be used to produce models that integrate gene expression changes across mutants to understand both the unique roles of each factor and crosstalk between signals. In Aim 2 we hypothesize that "core" sets of enhancers are responsible for the ERP for each signal, some of which mediate crosstalk between or feedback within a signaling pathway, as well as insulating pathways from one another. We will address this by isolating enhancers for genes identified in Aim 1 and testing their activities in both mice and zebrafish. These will be incorporated into our mathematical models to understand enhancer sensitivity and how this regulates sharpness of gene expression boundaries. Our long-term goal is to build a comprehensive model for a craniofacial gene regulatory network that can be amended as new data are available.

描述（由申请人提供）：大部分颅面骨骼源自咽弓，这些 3D 结构随着时间的推移在形状和基因表达方面经历复杂的变化。然而，尚未在体内对弓中的早期基因表达谱进行详细分析。如果没有这些信息，就不可能预测基因表达变化将如何影响随后的骨骼发育。我们的实验室广泛研究了足弓中的内皮素 1 (Edn1)、骨形态发生蛋白 (Bmp)、Wnt 和 Jag/Notch 信号通路，因为这四种通路塑造了面部骨骼的背腹 (D-V) 轴。我们已经表明，Edn1 和 Bmp 信号传导最初促进腹侧表达的基因，然后将拱细分为单独的 D-V 子域。我们的初步数据表明，Wnt 信号传导控制对 Edn1/Bmp 的反应能力，并且这三种途径与 Jag/Notch 信号传导相反。这四个信号调节的通路是高度动态的，包含多个反馈回路和串扰，从而创建了一个强大的抗扰动系统。通过收集所有四种信号的小鼠和斑马鱼突变体，我们处于独特的地位，可以足够详细地评估跨物种基因表达的保守性，以进行计算建模。我们的目标是使用这些模型来预测在这些信号的遗传扰动后观察到的硅面部缺陷。我们的双物种方法将识别新的候选基因并生成与人类颅面遗传学临床相关的模型。为了实现这些目标，我们将追求两个具体目标。在目标 1 中，我们假设虽然 Edn1、Bmp、Wnt 和 Jag/Notch 信号对于建立沿 D-V 轴的足弓中骨骼祖细胞的初始身份都至关重要，但它们各自发挥着不同的作用。我们将通过使用高通量 RNA 测序来定义所有四种途径突变体中基因表达的早期变化来解决这一假设。这些早期反应谱 (ERP) 将用于生成整合突变体基因表达变化的模型，以了解每个因子的独特作用以及信号之间的串扰。在目标 2 中，我们假设增强子的“核心”组负责每个信号的 ERP，其中一些增强子介导信号通路之间的串扰或信号通路内的反馈，以及使通路彼此绝缘。我们将通过分离目标 1 中确定的基因的增强子并在两只小鼠中测试它们的活性来解决这个问题和斑马鱼。这些将被纳入我们的数学模型中，以了解增强子的敏感性以及它如何调节基因表达边界的锐度。我们的长期目标是建立一个颅面基因调控网络的综合模型，该模型可以随着新数据的出现而进行修改。