Fox Transcription Factors in Development of Pulmonary Capillaries

肺毛细血管发育中的 Fox 转录因子

• 批准号: 7799820
• 负责人: Vladimir Kalinichenko
• 金额: $ 36.41万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7799820
• 关键词: Address; Affect; Alleles; Alveolar; Alveolus; Apoptosis; Applications Grants; Binding; Binding Sites; Blood Vessels; Blood capillaries; Boxing; Breeding; Capillary Endothelial Cell; Cell Differentiation process; Cell Proliferation; Cells; Childhood; Complementary DNA; Defect; Development; Diagnosis; Embryo; Embryonic Development; Embryonic Heart; Endoderm; Endothelial Cells; Enhancers; Epithelial Cells; Event; Exhibits; Family; Foxes; Gases; Gene Expression; Gene Targeting; Genes; Genetic; Genetic Screening; Genetic Transcription; Hemorrhage; Human; Laminin; Liver; Lung; Lung diseases; Mediating; Mesenchymal; Mesoderm; Mesoderm Cell; Messenger RNA; Morphogenesis; Mus; Newborn Infant; Normalcy; Organ; Pathway interactions; Perinatal; Peripheral; Phenotype; Play; Polyploid Cells; Primitive foregut structure; Promoter Regions; Proteins; Publishing; Pulmonary alveolar structure; Regulatory Pathway; Research Personnel; Role; Severities; Signal Pathway; Signal Transduction; Specialized Epithelial Cell; Staging; Tamoxifen; Testing; Time; Transgenes; Transgenic Mice; Vascular Endothelial Growth Factor Receptor; aurora B kinase; base; capillary; capillary bed; cell type; human PLK1 protein; laminin-1; lung Kruppel-like factor; lung development; lung injury; lung lobe; member; notch protein; novel; premature; prevent; programs; promoter; receptor; receptor-mediated signaling; recombinase; respiratory distress syndrome; transcription factor

DESCRIPTION (provided by applicant): Forkhead Box (Fox) transcription factors Foxml and Foxfl are expressed in developing pulmonary endothelial cells and play an essential role in lung development. Deficiency of either one of these Fox genes is associated with severe defects in formation of peripheral pulmonary capillaries as well as abnormalities in many organs causing embryonic lethality. Our hypothesis is that Foxml and Foxfl regulate transcription of genes critical for proliferation and differentiation of endothelial cells during pulmonary vascular development. In order to test this hypothesis, we will use the Tie2 promoter driven Tamoxifen-inducible Cre recombinase (Tie2-Cre-ERT2) transgene to mediate developmental-stage-specific deletion of either Foxfl or Foxml LoxP/LoxP (fl/fl) targeted allele in endothelial cells. In Aim 1, we describe plans to use Tie2-Cre-ERT2 Foxml fl/fl embryonic lungs and cultured pulmonary endothelial cells derived from these embryos to determine whether Foxml is required for endothelial cell proliferation and differentiation during lung vascular development. In Aim 2, we will develop a mouse containing a Foxfl fl/fl targeted allele and use the Tie2-Cre- ERT2 transgene to mediate temporal endothelial-specific deletion of the Foxfl fl/fl at different stages of lung development. We will use Tie2-Cre-ERT2 Foxfl fl/fl mice to determine whether Foxfl directly regulates endothelial cell differentiation and identify a time period during which Foxfl function is required to elicit proper vascular lung development and to regulate Foxfl target genes. We have shown that Foxfl regulates expression of genes involved in Notch-2 signaling and that the Notch-2 pathway is critical for normal development of lung microvasculature. In Aim 3, we propose to generate mice with the endothelial-specific Tie-2 promoter driven Notch-2 transgene. These Tie-2 Notch-2 mice will be bred with Foxfl mice to determine whether increased Notch-2 receptor levels in embryonic Foxfl lung will restore normalcy to the development of alveolar microvasculature and prevent perinatal pulmonary hemorrhage in Foxfl newborn mice. Completion of the proposed studies will enable us to determine the role of Foxfl and Foxml proteins in endothelial cells during lung development. These Fox proteins may also provide novel targets in genetic screening, diagnosis and treatment of human pediatric lung diseases including perinatal pulmonary hemorrhage and respiratory distress syndrome of premature born babies.

描述（申请人提供）：Forkhead Box （Fox）转录因子Foxml和Foxfl在肺内皮细胞发育过程中表达，在肺发育过程中起重要作用。缺乏这些Fox基因中的任何一个都与周围肺毛细血管形成的严重缺陷以及许多器官的异常有关，导致胚胎死亡。我们的假设是Foxml和Foxfl调节肺血管发育过程中内皮细胞增殖和分化的关键基因的转录。为了验证这一假设，我们将使用Tie2启动子驱动的他莫昔芬诱导的Cre重组酶（Tie2-Cre- ert2）转基因来介导内皮细胞中Foxfl或Foxml LoxP/LoxP （fl/fl）靶向等位基因的发育阶段特异性缺失。在Aim 1中，我们描述了使用Tie2-Cre-ERT2 Foxml fl/fl胚胎肺和来自这些胚胎的培养肺内皮细胞来确定在肺血管发育过程中内皮细胞增殖和分化是否需要Foxml的计划。在Aim 2中，我们将培养含有Foxfl /fl靶向等位基因的小鼠，并使用Tie2-Cre- ERT2转基因介导Foxfl /fl在肺发育的不同阶段的时间内皮特异性缺失。我们将使用Tie2-Cre-ERT2 Foxfl /fl小鼠来确定Foxfl是否直接调节内皮细胞分化，并确定Foxfl功能诱导血管肺发育和调节Foxfl靶基因所需的时间段。我们已经证明Foxfl调节Notch-2信号通路相关基因的表达，并且Notch-2通路对肺微血管的正常发育至关重要。在Aim 3中，我们提出用内皮特异性Tie-2启动子驱动的Notch-2转基因小鼠。这些Tie-2 Notch-2小鼠将与Foxfl小鼠杂交，以确定Foxfl胚胎肺中Notch-2受体水平的升高是否会恢复Foxfl新生小鼠肺泡微血管的正常发育并预防围产期肺出血。完成这些研究将使我们能够确定Foxfl和Foxml蛋白在肺发育过程中内皮细胞中的作用。这些Fox蛋白也可能为围产期肺出血和早产儿呼吸窘迫综合征等人类儿科肺部疾病的遗传筛查、诊断和治疗提供新的靶点。