Transcriptional regulatory landscapes underlying FEZ Formation

自由经济区形成的转录监管格局

基本信息

批准号：
10216391
负责人：
RALPH S MARCUCIO
金额：
$ 4.57万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10216391
关键词：
ATAC-seq B-Cell Acute Lymphoblastic Leukemia Binding Biological Assay Birds Brain Catalogs Cells ChIP-seq Chick Embryo Chromatin Cleft Lip Cleft Palate Competence Complex Congenital Abnormality Craniofacial Abnormalities Data Data Set Development Developmental Biology Disease Dysmorphology Ectoderm Ectoderm Cell Embryo Enhancers Etiology Excision Exhibits Face Fibroblast Growth Factor 8 Foundations Gene Expression Gene Expression Regulation Genes Genetic Genetic Transcription Genomic approach Goals Growth Holoprosencephaly Homeobox Human Jaw Kinetics Knowledge Laboratories Light Limb structure Location Maintenance Molecular Morphogenesis Morphology Mus Neural Crest Neural Crest Cell Nose Organ Outcome PBX3 gene Patients Pattern Process Prosencephalon Regulation Regulator Genes Regulatory Element Research Role SHH gene Shapes Signal Transduction Solid Sonic Hedgehog Pathway Structure Surface Ectoderm Testing Time Tissues Work base cell motility cleft lip and palate craniofacial craniofacial complex design experimental study genetic variant global run on sequencing homeodomain innovation loss of function member neurotransmission response risk variant smoothened signaling pathway transcription factor

项目摘要

Summary: Morphogenesis of vertebrate tissues and organs occurs through well-orchestrated tissue interactions. During development of the upper jaw, signaling interactions among the forebrain, the facial ectoderm, and the intervening neural crest cells regulate development of the upper jaw. Our research has revealed that signals from the brain and the neural crest cells act together to induce expression of Sonic hedgehog (SHH) in the Frontonasal Ectodermal Zone (FEZ), a signaling center located in the surface ectoderm of the Frontonasal Process (FNP) that directs patterned growth of the upper jaw anlagen. Our preliminary data also suggest that Pre-B-cell leukemia homeobox (PBX) transcription factors are involved in inducing and maintaining the pattern of SHH expression in the FEZ by an interacting genetic network. The research proposed in this application is designed to uncover the gene regulatory mechanisms that are activated by signals from the forebrain and the neural crest, as well as, to investigate the role of PBX transcription factors in regulating SHH expression in the FEZ. Specifically, we hypothesize that 1) SHH signaling from the forebrain induces “competence” in surface ectoderm cells to express SHH; 2) subsequently, upon arrival, neural crest cells induce transcription of SHH in competent cells of the FEZ, and 3) PBX transcription factors participate in regulating expression of SHH in the FEZ. We will test this hypothesis in three specific aims using avian and murine embryos. In each Aim, we will use chick embryos because they allow us to manipulate the signaling interactions between the brain and the FEZ (Aim 1), the neural crest cells and the ectoderm (Aim 2), or expression of PBX1 and PBX3 (Aim 3). We will use a variety of genomic approaches (ATAC-seq, ChIP-seq, GRO-seq) to evaluate changes in chromatin and activated transcriptional regulatory elements (TREs) at the SHH locus. In Aim 3, we will also assess morphological, cellular, and molecular outcomes of altering PBX gene expression, and we will use mouse embryos to take advantage of data already generated in the Selleri laboratory. The results of the work proposed in this application will shed light on the molecular aspects of facial development, and will aid understanding of disease processes that occur in this region. Allelic variants in SHH, or SHH pathway members, are associated with human dysmorphologies of the craniofacial complex in diseases such as Holoprosencephaly and cleft lip with our without cleft palate. Recently, PBX genes have been identified as risk variants in patients with craniofacial dysmorphology. Hence, our innovative approach combining studies with a solid foundation in developmental biology with chromatin analyses will yield information that is directly applicable to understanding human dysmorphology.

概括：脊椎动物组织和器官的形态发生是通过精心策划的组织相互作用发生的。上颌的发育，前脑、面部外胚层和下颌之间的信号相互作用我们的研究表明，干预神经嵴细胞调节上颌的发育。来自大脑的神经嵴细胞共同作用，诱导 Sonic Hedgehog (SHH) 在额鼻外胚层区 (FEZ)，位于额鼻表面外胚层的信号中心我们的初步数据还表明，指导上颌原基图案化生长的过程（FNP）。前 B 细胞白血病同源盒 (PBX) 转录因子参与诱导和维持该模式通过与遗传网络相互作用来控制 FEZ 中 SHH 的表达本申请提出的研究是旨在揭示由前脑和大脑信号激活的基因调控机制神经嵴，以及研究 PBX 转录因子在调节 SHH 表达中的作用具体来说，我们认为 1) 来自前脑的 SHH 信号诱导表面的“能力”。外胚层细胞表达 SHH；2) 随后，神经嵴细胞到达后诱导 SHH 转录 3) PBX 转录因子参与调节 FEZ 感受态细胞中 SHH 的表达 FEZ。我们将使用鸟类和小鼠胚胎在三个特定目标中测试这一假设。使用鸡胚胎，因为它们使我们能够操纵大脑和大脑之间的信号相互作用 FEZ（目标 1）、神经嵴细胞和外胚层（目标 2）或 PBX1 和 PBX3 的表达（目标 3）。将使用各种基因组方法（ATAC-seq、ChIP-seq、GRO-seq）来评估染色质的变化在目标 3 中，我们还将评估 SHH 基因座处的激活转录调控元件 (TRE)。改变 PBX 基因表达的形态、细胞和分子结果，我们将使用小鼠胚胎利用 Selleri 实验室已经生成的数据。本申请中提出的方法将阐明面部发育的分子方面，并将有助于了解该区域发生的疾病过程 SHH 或 SHH 通路中的等位基因变异。成员，与以下疾病中的人类颅面复合体畸形有关最近，PBX 基因已被确定为前脑无裂畸形和唇裂（伴有腭裂）的风险。因此，我们将研究与颅面畸形患者的变异相结合。通过染色质分析奠定发育生物学的坚实基础将产生直接的信息适用于理解人类畸形。