Molecular mechanisms driving cessation of neural crest migration and aggregation into cranial ganglia

驱动神经嵴迁移和聚集到颅神经节停止的分子机制

基本信息

批准号：
10309332
负责人：
Hugo Alexander Urrutia
金额：
$ 4.68万
依托单位：
CALIFORNIA INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10309332
关键词：
ATAC-seq Address Adhesions Automobile Driving Behavior Binding Sites Cadherins Candidate Disease Gene Cell Adhesion Molecules Cells Cephalic Characteristics Cytoskeleton Data Set Destinations Development Dorsal Ectopic Expression Elements Embryo Enhancers Epithelial Familial Dysautonomia Fibrosis Fluorescence-Activated Cell Sorting Ganglia Gene Expression Genes Genetic Transcription Genome Goals Health In Situ Hybridization Knowledge Lead Light Maintenance Malignant Neoplasms Mediating Mesenchymal Molecular Neural Crest Neural Crest Cell Neural tube Neuraxis Neuroblastoma Peripheral Peripheral Nervous System Phase Pheochromocytoma Physical condensation Pigments Play Population Process Reaction Regulator Genes Regulatory Element Reporter Role Skeleton Structure Structure of trigeminal ganglion Tissues Transcript Transcriptional Regulation Untranslated RNA Vertebrates XCL1 gene cell type craniofacial epithelial to mesenchymal transition experimental study gastrulation genome-wide in vivo insight loss of function melanoma migration neuromechanism novel somitogenesis spatiotemporal stem-like cell targeted treatment transcription factor transcriptome transcriptome sequencing tumor progression wound healing

项目摘要

Proposal Summary Neural crest cells are an important stem-like cell population characterized by their multipotency and migratory ability. Originating within the forming central nervous system, neural crest cells undergo a spatiotemporally regulated epithelial-to-mesenchymal transition (EMT) to leave the neural tube and become migratory. They then migrate extensively throughout the developing embryo, giving rise to a wide range of derivatives as diverse as elements of the craniofacial skeleton and peripheral nervous system. In the post- migratory phase, neural crest cells condense into different structures, a process that involves loss of migratory characteristics, perhaps reflecting the reverse of the EMT process. While neural crest EMT has been studied extensively, the mechanisms underlying the condensation of neural crest cells to form final derivatives is far less well characterized. To address this knowledge gap, we propose to identify transcriptional changes that occur during gangliogenesis with the goal of identifying those mediating alterations in intercellular adhesion required for neural crest condensation into peripheral ganglia. Our hypothesis is that the gene regulatory mechanisms that play a role during peripheral ganglion formation may reflect a reversal of the EMT process. The goal is to uncover the molecular mechanisms that drive condensation of neural crest cells into ganglia. These may in turn lead to clues regarding the underlying cause of certain types of neurocristopathies like familial dysautonomia and neural crest-derived cancers like neuroblastoma and pheochromocytoma. Aim 1: RNA-sequencing of pure populations of post-migratory cranial neural crest cells: RNA-sequencing of isolated condensing cranial neural crest cells will allow us to identify novel transcription factors and adhesion molecules that may drive neural crest condensation into cranial ganglia. Aim 2: Functional analysis of genes selectively upregulated upon condensation to form ganglia: Identified upregulated genes in condensing cranial neural crest cells will be validated by in situ hybridization and Hybridization Chain Reaction. We will then perform systematic loss-of-function and ectopic expression experiments on selected genes to examine their role in regulating condensation into and differentiation of peripheral ganglia. Aim 3: Characterization of cis-regulatory elements modulating gene expression during ganglion condensation: To identify putative enhancers driving gene expression during cranial neural crest condensation, we will perform ATAC-sequencing to identify conserved noncoding regions in the genome that are accessible to transcription factors during cranial neural crest condensation.

提案摘要神经嵴细胞是一种重要的干细胞样细胞群，其特征在于其多能性和迁徙能力。神经嵴细胞起源于正在形成的中枢神经系统，经历了时空调节的上皮间质转化（EMT）离开神经管并成为迁徙的。然后它们在整个发育胚胎中广泛迁移，产生多种其衍生物与颅面骨骼和周围神经系统的元素一样多样。在后- 迁移阶段，神经嵴细胞凝结成不同的结构，这个过程涉及迁移能力的丧失特征，或许反映了 EMT 过程的相反过程。虽然神经嵴 EMT 已被研究从广泛意义上讲，神经嵴细胞凝聚形成最终衍生物的机制要少得多特征良好。为了解决这一知识差距，我们建议识别转录变化发生在神经节生成过程中，目的是识别那些介导细胞间质改变的神经嵴凝结成周围神经节所需的粘附力。我们的假设是基因在外周神经节形成过程中发挥作用的调节机制可能反映了 EMT 的逆转过程。目标是揭示驱动神经嵴细胞凝结成的分子机制神经节。这些反过来可能会导致有关某些类型的神经脆病的根本原因的线索例如家族性自主神经功能障碍和神经嵴衍生的癌症，例如神经母细胞瘤和嗜铬细胞瘤。目标 1：对迁移后颅神经嵴细胞纯群体进行 RNA 测序：RNA 测序分离的浓缩颅神经嵴细胞将使我们能够识别新的转录因子和粘附可能驱动神经嵴凝结成颅神经节的分子。目标 2：在凝结形成神经节时选择性上调的基因的功能分析：浓缩颅神经嵴细胞中鉴定出的上调基因将通过原位杂交和杂交链式反应。然后我们将进行系统的功能丧失和异位表达对选定的基因进行实验，以检查它们在调节凝结和分化中的作用周围神经节。目标 3：神经节期间调节基因表达的顺式调控元件的表征凝结：识别颅神经嵴凝结过程中驱动基因表达的推定增强子，我们将进行 ATAC 测序来识别基因组中可访问的保守非编码区域颅神经嵴凝结过程中的转录因子。