GRNs Guiding Cranial Versus Trunk Neural Crest Formation

GRN 指导颅神经嵴形成与躯干神经嵴形成

• 批准号: 8752121
• 负责人: Marianne Bronner
• 金额: $ 31.21万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8752121
• 关键词: Affect; Afferent Neurons; Automobile Driving; Bar Codes; Birds; Cartilage; Cell Lineage; Cells; Cephalic; Characteristics; Congenital Abnormality; Core Facility; Coupled; Data; Development; Dissection; Elements; Embryo; Endomesoderm; Enhancers; Face; Ganglia; Gene Expression Profile; Gene Order; Gene Targeting; Genes; Goals; Grant; Hand; Head; Information Networks; Laboratories; Malignant Neoplasms; Mediating; Methods; Modeling; Multipotent Stem Cells; Nature; Neural Crest; Neural Crest Cell; Neuroblastoma; Neuroglia; Neurons; Pathway interactions; Peripheral; Peripheral Nervous System; Population; Positioning Attribute; Regulator Genes; Reporter; Sea Urchins; Series; Signal Transduction; Signaling Molecule; Skeleton; Skin; Stem cells; System; Testing; Transplantation; base; blastomere structure; bone; combinatorial; comparative; craniofacial; egg; gene function; genome analysis; genome-wide; in vivo; loss of function; melanocyte; melanoma; nano-string; neural plate; new technology; novel; programs; receptor; relating to nervous system; research study; spinal nerve posterior root; stem; trait; transcription factor; tumor; vertebrate embryology

The neural crest is a multipotent embryonic cell population that contributes to diverse derivatives, including peripheral ganglia, cartilage and bone of the face, and melanocytes. We have proposed and tested a multistep gene regulatory network (GRN), comprised of a logical series of distinct regulatory steps that act in concert to imbue the cranial neural crest its defining traits. However, there are significant differences in developmental potential and migratory pathways of different neural crest populations arising at different axial levels. Here, we propose to explore GRN differences along the neural axis, focusing on premigratory neural crest cells from two distinct regions: cranial versus trunk. Our preliminary transcriptome analysis reveals many transcription factors and signaling molecules specific to the cranial but not trunk neural crest or vice versa. Our goal is to determine the position of these genes in the cranial versus trunk GRNs. This systems level strategy will provide understanding of why NC GRNs produces a particular regulatory state for use in preprogramming these cells to a different state. The aims are: Aim 1: Multiplex perturbation analysis of GRN connections at cranial and trunk levels. With the genomewide representation ofthe active transcriptome of premigratory cranial and trunk neural crest in hand, we will perform loss-of-function experiments to perturb gene function and quantitate subsequent global transcriptional changes in putative target genes in single embryos using Nanostring analysis. Aim 2: Phylogenomic and funcfional analysis/dissection of neural crest enhancers. We will identify cisregulatory elements that mediate expression of key GRN factors in cranial versus trunk neural crest populations. We will perform multidimensional modeling that incorporates results of transcriptome data and active enhancers with functional perturbation results into representational models of neural crest GRNs. Aim 3: Reengineering ofthe trunk neural crest program to test skeletogenic potenfial. Using GRN informafion, we will challenge the fate of trunk NC by reengineering their regulatory circuits and observing if misexpression/deletion of key GRN subcircuits affects their identify and ability to contribute to cartilage.

神经嵴是一个多能胚胎细胞群，有助于产生多种衍生物， 包括周围神经节、面部软骨和骨骼以及黑素细胞。我们已提出并 测试了多步基因调控网络（GRN），由一系列逻辑上不同的调控步骤组成 它们协同作用，赋予颅神经嵴其定义特征。然而，有显着 不同神经嵴群体的发育潜力和迁移途径的差异 不同的轴向水平。在这里，我们建议沿着神经轴探索 GRN 差异，重点关注 来自两个不同区域的迁移前神经嵴细胞：颅骨与躯干。我们的初步转录组 分析揭示了许多特定于颅骨而非躯干的转录因子和信号分子 神经嵴，反之亦然。我们的目标是确定这些基因在颅骨和躯干中的位置 GRN。该系统级策略将帮助您理解为什么 NC GRN 会产生特定的 用于将这些细胞预编程为不同状态的调节状态。目标是： 目标 1：颅骨和躯干水平 GRN 连接的多重扰动分析。随着全基因组 手头的迁徙前颅骨和躯干神经嵴的活性转录组的表示，我们将 进行功能丧失实验以扰乱基因功能并定量随后的全局 使用纳米线分析单个胚胎中假定目标基因的转录变化。 目标 2：神经嵴增强子的系统基因组学和功能分析/解剖。我们将确定顺监管 介导颅神经嵴和躯干神经嵴关键 GRN 因子表达的元件 人口。我们将进行多维建模，其中结合了转录组数据的结果和 具有功能扰动的主动增强子导致神经嵴 GRN 的代表性模型。 目标 3：重新设计躯干神经嵴程序以测试成骨潜力。使用GRN 信息，我们将通过重新设计其调节电路来挑战主干 NC 的命运，并观察是否 关键 GRN 子电路的错误表达/删除会影响其识别和促进软骨的能力。