Characterizing the neural crest response to BMP signaling through gastrulation and neurulation

通过原肠胚形成和神经形成表征神经嵴对 BMP 信号的反应

• 批准号: 10000879
• 负责人: Michael Louis Piacentino
• 金额: $ 16.36万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10000879
• 关键词: Binding; Biology; CHARGE syndrome; California; Cartilage; Cells; Cephalic; Cleft Palate; Clustered Regularly Interspaced Short Palindromic Repeats; Congenital Abnormality; Craniofacial Abnormalities; Data Set; Defect; Dentin; Development; DiGeorge Syndrome; Diffusion; Ectoderm; Embryo; Environment; Epidermis; Event; Face; Ganglia; Gastrula; Genes; Genetic Epistasis; Goals; Image; Individual; Inhibitor of Differentiation Proteins; Institutes; Instruction; Lateral; Lead; Ligands; Mandibulofacial Dysostosis; Measures; Medial; Mentors; Molecular; Neural Crest; Neural Crest Cell; Pathway interactions; Peptide Hydrolases; Peripheral; Peripheral Nervous System; Phase; Plant Roots; Play; Population; Reaction; Regulation; Reporter; Research; Role; Signal Pathway; Signal Transduction; Skeleton; System; Systems Biology; Technology; Testing; Therapeutic; Time; TimeLine; Tooth structure; Training; blastomere structure; body system; bone; bone morphogenetic protein receptors; career; cell type; chromatin immunoprecipitation; craniofacial; craniofacial development; craniofacial disorder; craniofacial structure; epithelial to mesenchymal transition; experience; experimental study; extracellular; gastrulation; in silico; in vivo; in vivo Model; loss of function; mathematical model; morphogens; neural plate; novel; prevent; programs; public health relevance; receptor expression; relating to nervous system; response; spatiotemporal; tool; transcriptome sequencing; translational approach

PROJECT SUMMARY The cranial neural crest (NC) contributes to the formation of many craniofacial structures including the bones and cartilage of the face, tooth dentin and peripheral ganglia. Cell signaling regulates different aspects of cranial NC specification, epithelial-to-mesenchymal transition (EMT) and differentiation and disruptions in this developmental program results in many cranial NC-derived craniofacial birth defects including craniosynestosis, Treacher Collins and CHARGE syndromes, and cleft palate. BMP signaling plays a crucial role during the specification and differentiation of cranial NC, and more recently, BMP signaling was shown to control cranial NC EMT. A mechanistic understanding of the role of BMP signaling during cranial NC development is essential to develop novel preventative and therapeutic measures against craniofacial defects. This proposal will determine the molecular mechanism of BMP gradient formation in the chick gastrula, and how this gradient regulates the formation of cranial cell types including neural, cranial NC, placode and epidermal fates. These experiments will use in vivo and in silico approaches to test the hypothesis that extracellular BMP ligands are produced primarily by the cranial NC and are actively shuttled over long distances to signal most strongly in the nonneural ectoderm. Next, quantitative expression analysis and live imaging will be used to establish the timeline of BMP signaling during gastrulation and neurulation, and analysis of the resulting datasets will determine population- and single-cell-level responses to BMP signals. Differences in signal timing and strength will then be correlated with direct input into different target genes. Finally, the role of BMP target genes Id1/2/3/4 and Fibin during cranial NC EMT will be investigated using in vivo functional analyses. Together, the results of these aims will provide a comprehensive understanding of the regulation and roles of BMP signaling events during early cranial NC development. In addition to identifying targets for translational avenues to prevent craniofacial birth defects, the mentored phase of this proposal will provide Dr. Michael Piacentino with necessary training as he prepares to begin his independent career. Dr. Marianne Bronner's lab at California Institute of Technology, and his assembled advisory council, provide the necessary tools, expertise, and training environment to efficiently execute the proposed aims and establish Dr. Piacentino's independence. This training will be instrumental as Dr. Piacentino begins his independent research program and will provide the experience needed to make lasting impacts on the field of BMP signaling during craniofacial development.

项目摘要 颅神经嵴（NC）有助于许多颅面结构的形成，包括 面部的骨骼和软骨、牙本质和外周神经节。细胞信号调节不同的方面 颅NC规格，上皮间质转化（EMT）和分化和中断， 该发育程序导致许多颅NC衍生的颅面出生缺陷， 颅缝早闭、Treacher柯林斯和CHARGE综合征以及腭裂。BMP信号在细胞内 在颅NC的规范和分化过程中的作用，最近，BMP信号被证明是 控制颅脑NC EMT。BMP信号在颅NC中作用的机制理解 发展是至关重要的，以发展新的预防和治疗措施，对颅面 缺陷 这一建议将确定骨形态发生蛋白梯度形成的分子机制在鸡 原肠胚，以及这种梯度如何调节颅细胞类型的形成，包括神经，颅NC， 基板和表皮的命运。这些实验将使用体内和计算机模拟方法来测试 假设细胞外BMP配体主要由颅侧NC产生， 在非神经外胚层中信号最强。接下来，定量表达分析 并且将使用实时成像来建立原肠胚形成和神经胚形成期间BMP信号传导的时间轴， 分析得到的数据集将确定群体和单细胞水平对BMP的反应 信号.然后，信号定时和强度的差异将与不同目标的直接输入相关联 基因.最后，将研究BMP靶基因Id 1/2/3/4和Fibin在颅NC EMT中的作用 使用体内功能分析。这些目标的结果将共同提供一个全面的 了解BMP信号事件在早期颅NC发育过程中的调节和作用。 除了确定预防颅面出生缺陷的转化途径的目标外， 本提案的指导阶段将为Michael Piacentino博士提供必要的培训， 开始他独立的职业生涯。玛丽安·布朗纳博士在加州理工学院的实验室， 组建咨询理事会，提供必要的工具、专业知识和培训环境， 执行所提出的目标，并建立Piacentino博士的独立性。这项培训将有助于 博士Piacentino开始他的独立研究计划，并将提供所需的经验，使 在颅面发育过程中对BMP信号传导领域的持久影响。