Hypoxia-Induced Mitogenic Factor in Perinatal Lung

描述(由申请人提供)： 这项研究的长期兴趣是了解和探索低氧诱导的有丝分裂因子(HIMF)在肺发育和成熟中的细胞和分子机制。HIMF是我们在低氧性肺动脉高压小鼠模型中发现的一种蛋白质。在肺部哮喘模型中，它也被称为FIZZ1(发现于炎症区)。HIMF属于一个新的富含半胱氨酸的细胞因子家族，已知在缺氧性和炎症性肺中特异表达。最重要的是，在围产期，HIMF在发育中的肺中高度上调。这项提案将集中在两个主要领域。首先，它将解决Himf基因在发育中的肺中的表达，特别是在围产期。其次，将探讨hif基因产物在肺发育成熟过程中的细胞和分子作用机制及其与支气管肺发育不良(BPD)和呼吸窘迫综合征(RDS)的关系。第一个目标将解决这一假设，即HIMF在发育中的肺中表达，并调节肺血管的发育，控制实质肺的成熟，并协调血管形成和肺泡化。我们将利用组织学和分子生物学技术研究HIF在发育肺中的时空表达及其与血管生成和肺泡化的关系。第二个目的将检验HIMF在肺的成熟中起重要作用的假设；干扰HIMF基因的表达可能导致肺发育/成熟延迟和支气管肺发育不良(BPD)。本研究的目的是利用RNA干扰(RNAi)技术，在体外培养和移植(体内)胚胎和新生儿肺中，用HIF蛋白、HIMF中和抗体或HIMF基因敲除技术，确定HIMF在围产期血管形成和肺泡化中的作用。第三个目标将解决这样的假设，即在发育中的肺中，hif基因的表达受到转录因子的调控，包括C/EBPot和HIF-2a。它将利用全面的启动子-报告基因转染研究、EMSA、足迹和过度表达研究来确定在围产期对HIF基因表达至关重要的蛋白质-DNA相互作用。

DESCRIPTION (provided by applicant): The long-term interest of this research program is to understand and explore the cellular and molecular mechanisms of hypoxia-induced mitogenic factor (HIMF), a protein we found in a mouse model of hypoxia-induced pulmonary hypertension, in lung development and maturation. It was also called FIZZ1 (found in inflammatory zone) in an asthma model in the lung. HIMF belongs to a new family of cysteine rich cytokines known to be specifically expressed in hypoxic and inflammatory lung. Most importantly, HIMF is highly upregulated in the developing lung during the perinatal period. This proposal will focus on two major areas. First, it will address HIMF gene expression in the developing lung, especially during the perinatal period. Second, it will investigate the cellular and molecular mechanisms of action of HIMF gene product in lung development and maturation and its relation with bronchopulmonary dysplasia (BPD) and respiratory distress syndrome (RDS). The first aim will address the hypothesis that HIMF is expressed in the developing lung and regulates pulmonary vascular development, controls maturation of the parenchymal lung, and coordinates vascularization and alveolarization. We will investigate the temporal-spatial expression of HIMF in the developing lung and its relationship with vasculogenesis and alveolarization using histological and molecular biological techniques. The second aim will examine the hypothesis that HIMF plays an important role in the maturation of the lung; disrupting HIMF gene expression might result in delayed lung development/maturation, and bronchopulmonary dysplasia (BPD). This aim will define the roles of HIMF in vascularization and alveolarization during the perinatal period in cultured (in vitro) and transplanted (in vivo) embryonic and neonatal lungs treated with HIMF protein, HIMF neutralizing antibody, or HIMF gene knock down with RNA interference (RNAi) techniques. The third aim will address the hypothesis that HIMF gene expression in the developing lung is regulated by transcription factors, including C/EBPot and HIF-2a. It will utilize comprehensive promoter-reporter transfection studies, EMSA, footprinting, and overexpression studies to define the protein-DNA interactions critical to HIMF gene expression during the perinatal period.