Transcriptional Control of Liver Development

肝脏发育的转录控制

• 批准号: 7483709
• 负责人: Joshua R. Friedman
• 金额: $ 13.33万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-30 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7483709
• 关键词: Address; Alagille Syndrome; Albumins; Alleles; Biliary; Biliary Atresia; Binding; Binding Sites; Biological Assay; Candidate Disease Gene; Cells; Childhood; Complex; Data; Defect; Derivation procedure; Detection; Development; Diagnosis; Ductal; Embryo; Endoderm; Endothelial Cells; Epithelial Cells; Family; Family member; Fluorescence-Activated Cell Sorting; Foundations; Future; Gene Expression; Gene Targeting; Genes; Genetic; Goals; Green Fluorescent Proteins; Growth; Hepatic; Hepatic artery; Hepatocyte; Human; Investigation; Keratin-19; Laboratories; Lead; Liver; Liver diseases; Measures; Mesenchyme; Messenger RNA; Microarray Analysis; Molecular; Molecular Profiling; Morphology; Mus; Nature; Nucleic Acid Regulatory Sequences; Numbers; Organ; Partner in relationship; Phase; Phenotype; Physicians; Play; Polymerase Chain Reaction; Portal triad; Portal vein structure; Prealbumin; Prevention; Primitive foregut structure; Principal Investigator; Process; Proliferating; Proteins; Purpose; Reagent; Regulation; Reporter; Reporter Genes; Research; Research Personnel; Research Training; Reverse Transcription; Role; Role playing therapy; Scientist; Signal Transduction; Signaling Molecule; Staging; Structure; Structure of septum transversum; Techniques; Testing; Time; Training; Transcriptional Regulation; Transfection; Transgenes; Transgenic Mice; Winged Helix; Work; alpha-Fetoproteins; base; bile duct; career; chromatin immunoprecipitation; improved; in vivo; liver function; member; mutant; novel; programs; promoter; protein expression; recombinase; skills; tool; transcription factor

DESCRIPTION (provided by applicant): The liver develops from cells of the ventral foregut endoderm which differentiate into heptoblasts and proliferate into the adjacent mesenchyme to form the liver bud. Most of these cells mature into hepatocytes, but some of those surrounding the portal veins develop into the ductal plate, which gives rise to the bile ducts. In mice, this is followed by the formation of the hepatic arteries, thereby completing the portal triad. Defects in hepatogenesis are responsible for several pediatric liver diseases. This proposal aims to train the principal investigator (PI) for a research career focused on the study of liver development with the ultimate purpose of ameliorating these and other liver disorders. The goal of Specific Aims 1-3 is to characterize the regulatory networks controlling hepatogenesis. The Foxa family of winged helix transcription factors is expressed in the endoderm immediately before the onset of hepatogenesis, and they bind to the regulatory regions of a number of liver-specific genes. The first specific aim tests the hypothesis that Foxa1 and Foxa2 play a central role in the specification and further development of the liver. Mouse embryos lacking Foxa1 and Foxa2 throughout the endoderm will be generated; preliminary results indicate that in the absence of these factors, hepatic specification does not occur. The mechanism of Foxa function in liver development will be explored using protein and mRNA detection techniques to identify liver transcription factors whose expression is dependent on Foxa1 or Foxa2. In the second specific aim, these findings will be extended using chromatin immunoprecipitation combined with promoter microarray analysis to identify novel Foxa1 and Foxa2 target genes. The third specific aim will expand the focus to include the later stages of bile duct and hepatic artery development through the derivation of a transgenic mouse line allowing the analysis of ductal plate and biliary epithelial cell-specific gene expression. This data will be used to test the hypothesis that hepatic arteriogenesis requires inductive signals from developing bile ducts. Over the course of these studies, the PI will acquire laboratory skills, research training, and critical reagents which will enable him to begin work as an independent physician-scientist. The information which will be obtained from this and future investigations will improve our understanding of liver development and may enhance the diagnosis, treatment, and prevention of liver disease in humans.

描述（由申请人提供）： 肝脏从腹侧内胚层的细胞中发展，该细胞分化为七生细胞并扩散到相邻的间质中形成肝芽。这些细胞中的大多数成熟成肝细胞，但是门静脉周围的一些细胞会发展到导管板中，这会导致胆管。在小鼠中，接下来是肝动脉的形成，从而完成了门户三合会。肝病发生缺陷是多种儿科肝病的原因。该建议旨在培训主要研究者（PI），以研究肝脏发育研究的研究生涯，最终目的是改善这些和其他肝脏疾病。 特定目标1-3的目的是表征控制肝脏发生的调节网络。在肝病发生之前，在内胚层中表达了有翼螺旋转录因子的FOXA家族，它们与许多肝特异性基因的调节区域结合。第一个特定目的检验了FOXA1和FOXA2在肝脏的规范和进一步发展中起着核心作用的假设。将生成整个内胚层中缺少FOXA1和FOXA2的小鼠胚胎；初步结果表明，在没有这些因素的情况下，不会发生肝规范。 FOXA功能在肝发育中的机制将使用蛋白质和mRNA检测技术探索，以鉴定其表达取决于FOXA1或FOXA2的肝转录因子。在第二个特定目的中，这些发现将使用染色质免疫沉淀与启动子微阵列分析相结合，以鉴定新颖的FOXA1和FOXA2靶基因。第三个特定的目标将扩大重点，以包括胆管和肝动脉发育的后期，通过推导转基因小鼠系，从而分析导管板和胆道上皮细胞特异性基因表达。该数据将用于检验以下假设：肝动脉生成需要发育中的胆管感应信号。 在这些研究过程中，PI将获得实验室技能，研究培训和关键试剂，这将使他能够开始担任独立的医师科学家。将从此和未来的研究中获得的信息将提高我们对肝脏发育的理解，并可能增强人类肝病的诊断，治疗和预防。