Systems Analysis of BMP Regulation in Developing Zebrafish Embryos

斑马鱼胚胎发育中 BMP 调控的系统分析

• 批准号: 8560753
• 负责人: David Michael Umulis
• 金额: $ 30.92万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-09 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8560753
• 关键词: Affect; Biochemical; Bone Morphogenetic Proteins; Cell Differentiation process; Cell Nucleus; Cells; Complex; Data; Data Sources; Development; Dorsal; Embryo; Experimental Designs; Family; Feedback; Gastrula; Gene Expression Profile; Gene Expression Profiling; Genes; Goals; Image; Image Analysis; Individual; Injury; Knowledge; Measurement; Measures; Mediating; Methods; Modeling; Molecular; Neoplasm Metastasis; Organogenesis; Outcome; Pathway interactions; Pattern; Pattern Formation; Reference Standards; Regulation; Role; Signal Transduction; Specific qualifier value; Stem cells; System; Systems Analysis; Testing; Time; Tissues; Vertebrates; Zebrafish; angiogenesis; base; blastocyst; bone morphogenetic protein 5; cell growth; cell type; chordin; data integration; design; disorder control; extracellular; fitness; gastrulation; human disease; information processing; mathematical model; model development; morphogens; mutant; predictive modeling; prospective; protein distribution; public health relevance; research study; spatiotemporal; tissue regeneration

DESCRIPTION (provided by applicant): Bone Morphogenetic Proteins (BMPs) act in developmental pattern formation as a paradigm of extracellular information that is spatially distributed in a gradient as a morphogen, specifying distinct cell types via morphogen levels. In vertebrate dorsal-ventral (DV) axial pattern formation the function, molecular partners, and role of each BMP component are fairly well understood, but the mechanism by which the combined function of the components form a robust BMP gradient is complex and still poorly understood. The field is at an impasse to go beyond the current paradigms and solve the mechanism by which the extracellular factors and multiple feedback loops interact and regulate each other's' activity spatially and temporally to generate a gradient that patterns the embryo. The zebrafish system is sufficiently well defined now to allow effective and testable mathematical models to be generated that could move this field forward. Moreover, we know very little about how BMP regulators modulate the actual signaling gradient. The objective is to discover and discriminate mechanisms of BMP regulation by utilizing quantitative image acquisition and analysis, geometrically accurate mathematical models of early zebrafish embryo DV patterning, and mixed-quality constraint based optimization. In Aim 1 the spatiotemporal formation of the BMP signaling gradient will be quantitatively investigated by measuring phospho-Smad 1/5 levels IN TOTO in wild-type and BMP component mutant zebrafish embryos, segment all nuclei in each embryo, and register the data to a standard embryo. These studies will provide the first-ever (semi)-quantitative data that can be used to discern the spatial and quantitative differences and similarities of individual BMP extracellular modulators to understand their roles in BMP signaling gradient formation. In Aim 2 networks for BMP-mediated signaling control will be identified by developing, optimizing, and analyzing 3D spatiotemporal models. An image-based zebrafish late blastula- gastrula embryo BMP pattern formation model will be developed and tested for multiple alternative mechanisms of BMP regulation that guide pattern formation dynamics. In aim 3 we will use Model-Based Optimal Design of Experiments to reduce the complexity of factorial design required for comprehensive analysis of multiple-component networks. Additionally, we will determine the mechanism of Cvl2, Tsg1, and Chd regulation of dynamic BMP signaling to test the model's predictive ability and delineate the action of this important network. The goal of this aim is to carry out simultaneous gene perturbation experiments that will provide the greatest amount of information pertaining to BMP regulation. Understanding the mechanism of BMP-mediated patterning in vertebrates will provide the basis for tightly controlling BMP signaling in tissue regeneration and other prospective treatments of human disease.

描述（由申请人提供）：骨形态发生蛋白（BMP）以发育模式形成起作用，作为细胞外信息的范式，在空间上以梯度分布为形态学，通过形态学水平指定了不同的细胞类型。在脊椎动物背腹侧（DV）轴向图案形成中，每个BMP成分的功能，分子伙伴和作用都得到了充分的理解，但是组件组合函数形成强大的BMP梯度的合并功能的机制是复杂的，并且仍然不懂。该领域正在超越当前范式，并解决细胞外因子和多个反馈回路相互作用的机制，并在空间和时间上互相调节彼此的活动，以生成一种梯度，以模拟胚胎。斑马鱼系统现在已经充分定义了，以允许生成有效且可测试的数学模型，以使该字段向前移动。此外，我们对BMP调节器如何调节实际信号梯度的了解一无所知。目的是通过利用定量图像采集和分析，早期斑马鱼胚胎DV构图的几何准确数学模型以及基于混合质量约束的优化来发现和区分BMP调节的机制。在AIM 1中，将通过测量野生型和BMP成分突变体斑马鱼胚胎中的磷酸化 - SMAD 1/5水平来定量研究BMP信号传导梯度的时空形成，并将每个胚胎中的所有核分段，并将数据注册到标准胚胎中。这些研究将提供有史以来的第一个（半）定量数据，可用于辨别单个BMP细胞外调节剂的空间和定量差异和相似性，以了解其在BMP信号梯度形成中的作用。在AIM 2网络中，将通过开发，优化和分析3D时空模型来确定用于BMP介导的信号控制的网络。将开发并测试基于图像的斑马鱼晚期囊肿胚胎BMP模式形成模型，并测试BMP调节的多种替代机制，以指导模式形成动力学。在AIM 3中，我们将使用基于模型的实验最佳设计来减少多组分网络的全面分析所需的阶乘设计的复杂性。此外，我们将确定动态BMP信号的CVL2，TSG1和CHD调节的机理，以测试模型的预测能力并描述该重要网络的作用。该目标的目的是进行同时提供基因扰动实验，以提供与BMP调节有关的最大信息。了解脊椎动物中BMP介导的图案的机制将为紧密控制组织再生中的BMP信号传导和其他前瞻性治疗人类疾病提供基础。