Ibuprofen and enteric neural crest precursor migration

布洛芬和肠神经嵴前体迁移

基本信息

批准号：
8917209
负责人：
Ellen Merrick Schill
金额：
$ 4.81万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-20 至 2016-09-19
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8917209
关键词：
Abdomen Actins Adhesions Affect Age Age-Years Anti-Inflammatory Agents Anti-inflammatory Biochemical Cause of Death Cell Survival Cells Cessation of life Child Colon Congenital Abnormality Congenital Megacolon Cytoskeleton Data Defect Development Diet Disease Distal Distant Enteral Enteric Nervous System Environmental Exposure Environmental Risk Factor Enzymes Exhibits Exposure to Family Fetal Development First Pregnancy Trimester Fishes Focal Adhesions Folic Acid Genes Genetic Predisposition to Disease Genetic Risk Goals Health Human Ibuprofen Image Immunohistochemistry In Vitro Incidence Individual Infant Intestines Investigation Lead Length Life Measures Mediating Medicine Microscopy Molecular Morbidity - disease rate Mus Mutation Nervous system structure Neural Crest Neural Tube Defects Neuroglia Neurons Obstruction PTGS1 gene PTGS2 gene Pathway interactions Pharmaceutical Preparations Physiology Population Preclinical Drug Evaluation Pregnancy Proteins Regulation Research Project Grants Risk Severities Structure Supplementation Susceptibility Gene Testing Therapeutic Time Transgenic Animals United States Vertebrates Vomiting Woman Zebrafish base cell motility cellular imaging critical period cyclooxygenase 1 disease-causing mutation disorder risk effective intervention gene environment interaction gene interaction improved in utero in vivo inhibitor/antagonist migration mortality mouse model nervous system development novel null mutation premature prevent research study time use

Abdomen Actins Adhesions Affect Age Age-Years Anti-Inflammatory Agents Anti-inflammatory Biochemical Cause of Death Cell Survival Cells Cessation of life Child Colon Congenital Abnormality Congenital Megacolon Cytoskeleton Data Defect Development Diet Disease Distal Distant Enteral Enteric Nervous System Environmental Exposure Environmental Risk Factor Enzymes Exhibits Exposure to Family Fetal Development First Pregnancy Trimester Fishes Focal Adhesions Folic Acid Genes Genetic Predisposition to Disease Genetic Risk Goals Health Human Ibuprofen Image Immunohistochemistry In Vitro Incidence Individual Infant Intestines Investigation Lead Length Life Measures Mediating Medicine Microscopy Molecular Morbidity - disease rate Mus Mutation Nervous system structure Neural Crest Neural Tube Defects Neuroglia Neurons Obstruction PTGS1 gene PTGS2 gene Pathway interactions Pharmaceutical Preparations Physiology Population Preclinical Drug Evaluation Pregnancy Proteins Regulation Research Project Grants Risk Severities Structure Supplementation Susceptibility Gene Testing Therapeutic Time Transgenic Animals United States Vertebrates Vomiting Woman Zebrafish base cell motility cellular imaging critical period cyclooxygenase 1 disease-causing mutation disorder risk effective intervention gene environment interaction gene interaction improved in utero in vivo inhibitor/antagonist migration mortality mouse model nervous system development novel null mutation premature prevent research study time use

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项目摘要

DESCRIPTION (provided by applicant): Birth defects are the most common cause of death in children under one year of age in the United States. The best way to reduce the morbidity and mortality of specific birth defects is to develop strategies to prevent them from occurring, as exemplified by the decrease of neural tube defects due to folic acid supplementation. Hirschsprung Disease (HSCR) is a birth defect of the enteric nervous system that occurs when enteric neural crest-derived cells (ENCCs) do not migrate all the way to the end of the bowel. The resulting distal bowel aganglionosis leads to a functional obstruction, which causes vomiting, abdominal distension and predisposes to premature death. There are many proteins important for normal ENCC migration and mutations in these genes increase the risk of HSCR, but none of these mutations cause disease in all affected children. Therefore, children who have HSCR must have additional factors that further increase their HSCR risk. To develop strategies to decrease HSCR occurrence, it is important to identify alterable factors, such as maternal diet or medications that could in combination with genetic defects cause disease in a child who would otherwise have been born healthy. Ibuprofen, a non-steroidal anti-inflammatory drug (NSAID) commonly used in pregnancy, inhibited ENCC migration in a zebrafish drug screen, causing a HSCR-like distal bowel aganglionosis in affected fish. Experiments in mammalian primary culture confirmed that ibuprofen slows ENCC migration and appears to disrupt actin cytoskeletal and focal adhesion regulation needed for efficient migration. The goals of this proposal are to determine the biochemical mechanism of ibuprofen's inhibition of ENCC migration and to determine if ibuprofen increases the likelihood of HSCR in genetically predisposed individuals. The biochemical mechanisms that ibuprofen disrupts to inhibit ENCC migration will be investigated using cutting edge time-lapse live cell imaging of fluorescently-tagged proteins. These investigations will allow for in depth analysis of ENCC actin protrusions and the dynamics of focal adhesions. In addition, ENCCs from transgenic animals with null mutations in Cox1 and Cox2, enzymes inhibited by ibuprofen, will be analyzed by immunohistochemistry and time-lapse imaging to determine if ibuprofen inhibits ENCC migration via a Cox- dependent mechanism. I will also determine if mice with null mutations in Cox1 and Cox2 have HSCR-like distal bowel aganglionosis. Finally, mice genetically predisposed to HSCR due to mutations in genes important for normal ENCC migration will be exposed to ibuprofen in utero during the critical period of ENS development. These experiments will examine if ibuprofen could be a common and avoidable exposure during pregnancy that increases HSCR risk by reducing the efficiency of ENCC migration.

描述（由申请人提供）：在美国一岁以下儿童中，先天缺陷是最常见的死亡原因。降低特定先天缺陷的发病率和死亡率的最佳方法是制定防止它们发生的策略，这是通过补充叶酸引起的神经管缺陷的减少来说明的。 Hirschsprung疾病（HSCR）是肠神经系统的先天缺陷，当肠神经rest衍生的细胞（ENCC）不会一直迁移到肠的尽头时。由此产生的远端肠道炎病导致功能性障碍物，导致呕吐，腹部延伸和易患性死亡。对于正常的ENCC迁移和这些基因的突变，有许多蛋白质会增加HSCR的风险，但是这些突变都没有引起所有受影响儿童的疾病。因此，患有HSCR的儿童必须有其他因素，以进一步增加其HSCR风险。为了制定减少HSCR发生的策略，重要的是要识别可改变的因素，例如孕产妇饮食或可能结合遗传缺陷的药物会导致一个否则将是健康的儿童疾病。布洛芬是一种非甾体类抗炎药（NSAID），通常在妊娠中使用，抑制了斑马鱼药物筛查中的ENCC迁移，在受影响的鱼中导致hscr样的消肠炎症性炎症。哺乳动物原发性培养的实验证实，布洛芬会减慢ENCC的迁移，并且似乎破坏了有效迁移所需的肌动蛋白细胞骨架和局灶性粘附调节。该提案的目标是确定布洛芬对ENCC迁移的抑制的生化机制，并确定布洛芬是否增加了基因易感性个体HSCR的可能性。布洛芬破坏了抑制ENCC迁移的生化机制将使用荧光标记蛋白的最新延时活细胞成像研究。这些研究将允许对ENCC肌动蛋白突起和局灶性粘连的动力学进行深入分析。此外，将通过免疫组织化学和延时成像对抑制的Cox1和Cox2中的转基因动物的ENCC进行抑制的酶，以确定Ibuprofen是否通过COX依赖机制抑制ENCC迁移。我还将确定COX1和COX2中无效突变的小鼠是否患有类似HSCR的远端肠动体动烟病。最后，由于ENS发育的关键时期，由于对正常ENCC迁移重要的基因的突变，由于对正常ENCC迁移重要的基因而导致的小鼠会暴露于子宫内布洛芬。这些实验将检查布洛芬在怀孕期间是否可能是常见且可避免的暴露，通过降低ENCC迁移效率来增加HSCR风险。