Transcriptional control of epithelial behaviors that drive mammalian neural tube closure

驱动哺乳动物神经管闭合的上皮行为的转录控制

• 批准号: 8887546
• 负责人: Lee A. Niswander
• 金额: $ 31.73万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8887546
• 关键词: Abbreviations; Actins; Address; Affect; Alleles; Animal Model; Attention; Binding; Binding Sites; Biology; Brain; CDH1 gene; Cell Adhesion; Cell Shape; Cell model; Cells; Cephalic; Characteristics; Child; Child Mortality; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Complex; Computer Simulation; Congenital Abnormality; Consensus; Coupling; DNA Binding; Defect; Distal; Dorsal; Drosophila genus; E-Cadherin; Ectoderm; Ectoderm Cell; Embryo; Embryonic Development; Emotional; Epithelial; Equilibrium; Ethylnitrosourea; Evaluation; Event; Exencephalies; Face; Failure; Family; Family member; Filopodia; Foundations; Gene Activation; Gene Expression Profile; Gene Expression Regulation; Gene Targeting; Genes; Genetic; Genetic Screening; Genetic Transcription; Genetic study; Goals; Health; Homologous Gene; Human; Human Genetics; Image; Knowledge; Lead; Life; MAPK8 gene; Malignant Neoplasms; Mediating; Membrane; Mesenchymal; Methods; Molecular; Morphogenesis; Mus; Mutation; Neural Crest; Neural Crest Cell; Neural Fold; Neural Tube Closure; Neural Tube Defects; Neural Tube Development; Neural tube; Neuraxis; Nucleotides; Pathway interactions; Pattern; Phenotype; Process; Public Health; Recycling; Reporter; Research; Role; Sampling; Signal Pathway; Signal Transduction; Site; Somatotropin-Releasing Hormone; Specificity; Spinal; Spinal Cord; Spinal Dysraphism; Surface Ectoderm; Technology; Testing; Time; Tissues; Tomatoes; Transcription Factor AP-1; Transcriptional Regulation; Translating; Tube; Variant; cancer therapy; cell behavior; cell growth regulation; critical developmental period; driving behavior; embryo tissue; epithelial to mesenchymal transition; fly; gene function; gene repression; insight; knock-down; meetings; mortality; mouse model; mutant; neural plate; new technology; novel; preference; prevent; programs; public health relevance; regional difference; relating to nervous system; repaired; research study; seal; transcription factor

 DESCRIPTION (provided by applicant): Failure of neural tube closure is a devastating birth defect. Research in the Niswander lab has provided significant insights into the molecular and cellular regulation of NT closure. We have created and studied mouse models with neural tube defects (NTDs) to elucidate the molecular foundations of NT closure. Moreover, we have created robust and novel technology to visualize NT closure in a living mammalian embryo. Our dynamic imaging and key genetic mutants have focused attention on the little studied but critical role for the non-neural ectoderm (NNE) in NT closure. In addition we developed methods to specifically isolate NNE cells to provide a refined and robust platform to study the biology of the NNE. Here we will build upon our unique perspective and turn our attention to spinal NTDs, to provide insight into the most common type of NTD in humans, and to two genetic pathways that are associated with spina bifida in mice and humans. Aim 1 will test the hypothesis that the two closely related Grainyhead-like (GRHL) transcription factors, GRHL2 and GRHL3, differentially control cranial and spinal NT closure through unique and differential activation of target genes, in part mediated by interaction with the JNK signaling pathway that activates the AP1 (cJUN/cFOS) transcription factor. Aim 2 will extend our live platform to test the hypothesis that GRHL-regulated NNE transcriptional programs drive NT closure by controlling cell adhesion, recycling of membrane components, cell shape changes, and/or actin dynamics. Aim 3 will combine our comprehensive molecular and cellular insights with novel unpublished analyses of hundreds of NTD samples to test the hypothesis that mutations identified in GRHL3 and the JNK pathway are causative for spinal NTDs in humans. Relevance of research to public health: The proposed experiments will lead to new cellular and molecular insights into the causes of caudal NTDs, the most common type of NTD and which leads to a profoundly important and frequently disrupted aspect of mammalian embryogenesis. Moreover, our studies will impart novel insights into the general mechanisms of embryonic tissue fusion including the face and body wall. The insights gained here may lead to therapies of general application for treatment of embryonic tissue closure defects that together represent a significant percentage of human birth defects. Abbreviations used in proposal: CDH1 Cadherin1 or E-cadherin EMT Epithelial-to-mesenchymal transition GRHL Grainyhead-like (GRH is the fly homolog) KD Knock-down mT/mG Membrane tomato/membrane GFP fluorescent reporter, GFP expression is activated by Cre NNE Non-neural or surface ectoderm NT Neural tube NTD Neural tube defect Grhl2-null: We will use Grhl21Nisw allele that we isolated in our ENU-screen and which has the same phenotype as other Grhl2 null alleles. Grhl3-Cre: We will use Grhl3-Cre which generates a null allele (obtained from S. Coughlin; Grhl3tm1(cre)Cgh).

 描述（由适用提供）：神经元管闭合的失败是毁灭性的先天缺陷。 Niswander Lab的研究为NT闭合的分子调节提供了重大见解。我们创建并研究了具有神经元管缺陷（NTD）的小鼠模型，以阐明NT闭合的分子基础。此外，我们创造了强大而新颖的技术，以可视化活着的哺乳动物胚胎中的NT闭合。我们的动态成像和关键的遗传突变体已将注意力集中在NT闭合中非神经外胚层（NNE）的较小但至关重要的作用上。此外，我们开发了特定分离NNE细胞的方法，以提供一个精致且可靠的平台来研究研究的生物学 nne。在这里，我们将基于我们独特的观点，并将注意力转向脊柱NTD，以深入了解人类中最常见的NTD类型，以及在小鼠和人类中与Spina Bifida相关的两种遗传途径。 AIM 1将检验以下假设：两个密切相关的颗粒状样（GRHL）转录因子GRHL2和GRHL3，通过与JNK信号通路相互作用介导的靶基因的唯一和差异激活，通过与JNK信号途径相互作用介导的AP1（CJUN/CFOS）的转录因子进行了差异地控制颅和脊柱NT闭合。 AIM 2将扩展我们的实时平台，以测试通过控制细胞粘附，膜组件的回收，细胞形状变化和/或肌动蛋白动力学来驱动NT闭合的假设。 AIM 3将将我们的综合分子和细胞见解与数百种NTD样品的新分析结合在一起，以测试以下假设：在GRHL3和JNK途径中鉴定出的突变对于人类的脊柱NTD是严重的。研究与公共卫生的相关性：拟议的实验将导致对尾骨NTD的原因的新细胞和分子见解，NTD是最常见的NTD类型，并导致哺乳动物胚胎发生的深刻重要且经常破坏的方面。此外，我们的研究将提供新的见解，以了解包括面部和身体壁在内的胚胎组织融合的一般机制。这里获得的见解可能会导致一般应用治疗胚胎组织闭合缺陷的疗法，这些缺陷共同代表了人类出生缺陷的很大一部分。 Abbreviations used in proposal: CDH1 Cadherin1 or E-cadherin EMT Epithelial-to-mesenchymal transition GRHL Grainyhead-like (GRH is the fly homolog) KD Knock-down mT/mG Membrane tomato/membrane GFP fluorescent reporter, GFP expression is activated by Cre NNE Non-neural or surface ectoderm NT Neural tube NTD Neural管缺陷grhl2-null：我们将使用我们在ENU屏幕中隔离的GRHL21NISW等位基因，并且与其他GRHL2 NULL等位基因具有相同的表型。 GRHL3-CRE：我们将使用生成无效等位基因的GRHL3-CRE（从S. coughlin; grhl3tm1（CRE）CGH获得）。