Mechanisms of folate action during nervous system development

叶酸在神经系统发育过程中的作用机制

基本信息

批准号：
10115144
负责人：
Laura Noemi Borodinsky
金额：
$ 34.34万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-05-15 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10115144
关键词：
Actins Adherens Junction Antibodies Apical Biological Assay C cadherin Cell Adhesion Cell Adhesion Molecules Cell Shape Cell-Cell Adhesion Cells Clinical Congenital Abnormality DNA Methylation DNA biosynthesis Diagnosis Embryo Endocytosis Endosomes Environmental Risk Factor Failure Fetal Development Fetus Folic Acid Genetic Genetic Epistasis Goals Human Image In Situ In Vitro Incidence Integral Membrane Protein Knowledge Measures Methodology Microscopy Microtubules Modification Molecular Myosin ATPase Nature Neural Tube Closure Neural Tube Defects Neural tube Neurons Newborn Infant Pathway interactions Phase Phenotype Phosphorylation Pregnancy Prevention Prevention Measures Protein Dynamics Proteins Proteomics Public Health Publishing Receptor Protein-Tyrosine Kinases Regulation Reporter Research Resolution Risk Role Signal Pathway Signal Transduction Signaling Molecule Supplementation Surface System Testing Time Ubiquitin Ubiquitination United States Western Blotting Xenopus laevis apical membrane base beta catenin constriction early pregnancy experimental study folate-binding protein folic acid supplementation fortification loss of function nervous system development neural plate neuroregulation novel optogenetics prevent rapid growth receptor recruit sensor trafficking ubiquitin ligase uptake

项目摘要

Project summary Neural tube defects (NTDs) are among the most common serious birth defects diagnosed in human fetuses and newborns with a combined incidence of ~1/1,000 in the United States and an estimated of 300,000 or more newborns worldwide each year. NTDs result from the failure of neural tube closure during the early fetal development. A combination of genetic and environmental factors appears to regulate the formation of the neural tube. Notably, folate supplementation during pregnancy prevents NTDs by unclear mechanisms. Our recently published study demonstrates that folate receptor 1 (Folr1), one of folate uptake systems, localizes to the apical surface of Xenopus laevis neural plate and is necessary for neural plate cell apical constriction during neural plate folding. Moreover, we find that Folr1 interacts with adherens junction components, C-cadherin and β- catenin suggesting that folate signaling might regulate neural plate cell-cell adhesion during neural tube formation. Our overall research goal is to elucidate the cellular and molecular mechanisms underlying neural tube formation. We will test the hypothesis that folate participates in the changes in cell shape that neural cells undergo during neurulation by recruiting its receptor and triggering a novel and dynamic signaling pathway. The first specific aim will consist in determining the molecular mechanisms underlying folate/Folr1 promotion of neural plate cell apical constriction during neural tube formation. We will identify the molecular mechanisms of Folr1 regulation of cell adhesion remodeling necessary for neural tube formation. In the second specific aim we will discover the signaling pathways recruited by folate/Folr1 that are necessary for neural plate cell apical constriction and neural tube formation. We will interrogate the ubiquitination pathway through gain and loss of function approaches and epistasis experiments. We will assess the role of folate in cell adhesion molecule and cytoskeletal dynamics by live imaging embryos expressing fluorescently tagged proteins or reporters of cell adhesion and cytoskeletal components during neural tube formation. We will use state-of-the-art methodologies including proteomics of immunoprecipitates, super resolution microscopy, reporters of cytoskeletal and cell adhesion dynamics and optogenetic approaches to manipulate signaling pathways. Although folate fortification has been a highly effective public health measure in reducing NTDs, the lack of mechanism-based understanding of NTD prevention leads to general concerns regarding unintended consequences resulting from supplementation. Optimal folate supplementation, risk groups and treatment of folate-insensitive NTDs are some of the unsolved clinical aspects awaiting for the full elucidation of the molecular and cellular mechanisms underlying folate action in neural tube formation.

项目摘要神经管缺陷（NTD）是在人类胎儿和在美国，新生儿总共发生了〜1/1,000，估计为30万或更多全世界新生儿。 NTD是由于胎儿早期闭合神经管的失败而导致的发展。遗传因素和环境因素的结合似乎调节了中性的形成管子。值得注意的是，怀孕期间的补充叶酸可以通过不清楚的机制来阻止NTD。我们最近已发表的研究表明，叶酸受体1（Folr1）是叶酸摄取系统之一，本地化于顶端爪蟾Laevis神经板的表面，对于神经元期间神经板细胞顶部收缩是必不可少的盘子折叠。此外，我们发现FOLR1与粘附连接成分C-钙粘着蛋白和β-相互作用 Catenin表明叶酸信号传导可能会调节神经管过程中神经铜细胞粘合剂形成。我们的总体研究目标是阐明中性的细胞和分子机制管形成。我们将测试叶酸参与细胞形状的变化的假设通过招募其接收器并触发新颖而动态的信号通路，在谈判过程中进行谈判。这第一个特定目的将包括确定叶酸/FELR1促进神经元的分子机制神经管形成期间的板细胞顶端收缩。我们将确定FOLR1的分子机制调节神经管形成所需的细胞粘附重塑。在第二个特定目标中，我们将发现叶酸/fly1募集的信号通路，这些信号通路是神经板细胞顶端所必需的收缩和神经管的形成。我们将通过收益和损失来询问泛素化途径功能方法和上位性实验。我们将评估叶酸在细胞粘合分子中的作用和通过实时成像胚胎表达荧光标记的蛋白质或细胞记者的细胞骨架动力学神经管形成过程中的粘附和细胞骨架成分。我们将使用最先进的方法包括免疫沉淀物的蛋白质组学，超级分辨率显微镜，细胞骨架和细胞的记者粘附动力学和光遗传学方法来操纵信号通路。虽然叶酸防御力缺乏基于机制的理解，一直是减少NTD的高效公共卫生措施 NTD的预防导致人们普遍担心补充。最佳叶酸补充，风险组和叶酸不敏感的NTD的治疗是某些在等待分子和细胞机制的尚未解决的临床方面神经管形成中的叶酸作用。