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rest（NC）有助于形成许多颅面结构，包括 脸部，牙齿牙本质和周围神经节的骨骼和软骨。细胞信号调节不同方面 颅NC规格，上皮到间质转变（EMT）以及分化和破坏 该发展计划导致许多颅内NC衍生的颅面出生缺陷，包括 颅内菌病，treacher柯林斯和电荷综合征和left裂。 BMP信号扮演至关重要 在颅NC的规范和分化中的作用，最近，BMP信号被证明是 控制颅NC EMT。对BMP信号在颅NC中的作用的机械理解 开发对于制定针对颅面的新型预防和治疗措施至关重要 缺陷。 该建议将确定雏鸡中BMP梯度形成的分子机制 胃，以及该梯度如何调节颅细胞类型的形成，包括神经，颅NC， 位置和表皮命运。这些实验将在体内和计算机方法中使用 假设细胞外BMP配体主要由颅NC产生，并积极穿梭 在非神经外胚层中最强烈的长距离发出信号。接下来，定量表达分析 现场成像将用于在胃和神经化过程中建立BMP信号的时间表 对所得数据集的分析将确定对BMP的种群和单细胞级响应 信号。然后，信号时序和强度的差异将与直接输入到不同目标相关 基因。最后，将研究BMP靶基因ID1/2/3/4和纤维NC EMT期间纤维的作用 使用体内功能分析。这些目标的结果一起将提供全面的 了解BMP信号事件在早期NC开发过程中的调节和作用。 除了确定转化途径的目标以防止颅面出生缺陷之外， 该提案的指导阶段将为迈克尔·皮亚森蒂诺（Michael Piacentino）博士提供必要的培训。 开始他的独立职业。加利福尼亚理工学院的Marianne Bronne博士实验室及其 组装咨询委员会，提供必要的工具，专业知识和培训环境以有效 执行拟议的目标并建立皮亚切诺博士的独立性。该培训将有助于 Piacentino博士开始了他的独立研究计划，并将提供制作所需的经验 对颅面发育过程中BMP信号传导领域的持久影响。