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：
10450672
负责人：
Hugo Alexander Urrutia
金额：
$ 4.76万
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/10450672
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rest细胞是一个重要的茎状细胞群，其特征是其多能量和迁移能力。起源于形成的中枢神经系统，神经rest细胞经历了时空调节的上皮到间质转变（EMT）离开神经管并变成迁移。然后，他们在整个发育中的胚胎中广泛迁移衍生物与颅面骨骼和周围神经系统的元素一样多样化。在邮政中 - 迁移阶段，神经rest细胞凝结成不同的结构，该过程涉及迁移的丧失特征，也许反映了EMT过程的相反。虽然已经研究了神经rest EMT 广泛的是，神经rest细胞形成最终衍生物的缩合的基础机制要少得多特征很好。为了解决这一知识差距，我们建议确定转录变化发生在神经节发生期间，目的是识别那些介导的细胞间变化神经rest缩合到周围神经节所需的粘附。我们的假设是基因在外围神经节形成期间起作用的调节机制可能反映EMT的逆转过程。目的是揭示将神经rest细胞缩合到进入的分子机制神经节。这些反过来可能会导致有关某些类型神经疾病的根本原因的线索像家族性动物障碍和神经rest衍生的癌症一样，如神经母细胞瘤和嗜铬细胞瘤。 AIM 1：媒介后颅神经rest细胞的RNA序列：RNA测序孤立的冷凝颅神经rest细胞将使我们能够鉴定新的转录因子和粘附可能将神经rest缩合到颅神经节的分子。目标2：在凝结后选择性上调以形成神经节的基因的功能分析：凝结颅神经rest细胞中发现的上调基因将通过原位杂交和杂交链反应。然后，我们将执行系统的功能丧失和异位表达对选定基因进行的实验，以检查其在调节凝结中的作用和分化外围神经节。目标3：在神经节期间调节基因表达的顺式调节元件的表征凝结：确定推定的增强子在颅神经c凝结过程中驱动基因表达，我们将执行ATAC序列，以识别基因组中可访问的保守的非编码区域，这些区域可访问颅神经rest缩合过程中的转录因子。