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：
10358628
负责人：
RALPH S MARCUCIO
金额：
$ 47.32万
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：
2020-03-11 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10358628
关键词：
ATAC-seq B-Cell Acute Lymphoblastic Leukemia Binding Biological Assay Birds Brain Catalogs Cells ChIP-seq Chick 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 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经过精心修整的组织相互作用发生的。期间上颌的发展，前脑之间的信号相互作用，面外胚层和干预神经rest细胞调节上颌的发育。我们的研究揭示了信号从大脑和神经rest细胞一起起作用，诱导声音刺猬（SHH）的表达额叶外胚层区域（FEZ），一个信号中心，位于额叶的表面外胚层过程（FNP）指导上颚Anlagen的图案增长。我们的初步数据也表明前B细胞白血病同型（PBX）转录因子参与诱导和维持模式通过相互作用的遗传网络在FEZ中表达的表达。该应用程序提出的研究是旨在发现由前脑和信号激活的基因调节机制神经波峰以及研究PBX转录因子在确定SHH表达中的作用 fez。特别是，我们假设1）来自前脑的SHH信号传导在表面上引起“能力” 外胚层细胞表达SHH； 2）随后，到达后，神经rest细胞诱导SHH的转录 FEZ的有能力的细胞和3）参与确定SHH表达的PBX转录因子 fez。我们将使用禽和鼠类胚胎在三个特定目标中检验这一假设。在每个目标中，我们都会使用小鸡胚胎，因为它们允许我们操纵大脑与大脑之间的信号传导相互作用 FEZ（AIM 1），神经rest细胞和外胚层（AIM 2）或PBX1和PBX3的表达（AIM 3）。我们将使用多种基因组方法（ATAC-SEQ，CHIP-SEQ，GRO-SEQ）评估染色质的变化并在SHH基因座处活化的转录调节元件（TRE）。在AIM 3中，我们还将评估改变PBX基因表达的形态学，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。工作结果在本应用中提出的将阐明面部发育的分子方面，并将有助于了解该地区发生的疾病过程。 SHH或SHH途径的等位基因变体成员与颅面复合物的人类运动障碍有关，例如全脑脑和唇裂，没有left裂。最近，PBX基因已被确定为风险颅面畸形患者的变体。因此，我们的创新方法将研究与通过染色质分析的发育生物学中的坚实基础将产生直接的信息适用于了解人类畸形学。