MUC 2 GENE ACTIVATION IN DEVELOPMENT AND DISEASE

慢性阻塞性呼吸道疾病，如囊性纤维化 以粘蛋白过度生产为特征的。随着时间的推移，这可能会导致 粘液堵塞的形成、感染、支气管扩张、呼吸道 功能不全，和死亡。理解这一系列事件是如何 被触发将有助于设计抑制它的方法。在.期间 上一阶段，我们的实验证实了一个假设，即早期事件 在实验中诱导的高分泌是稳定的 状态粘蛋白(MUC-2)m RNA(内毒素滴注诱导) 航空公司)。我们的实验还显示了MUC-2的阶段性上调 在气管发育的早期。拟议研究的目标是获得 有关如何触发和控制这些事件的信息。研究 在特定目标清单下，L将使用原位杂交技术确定， 气管发育过程中MUC-2激活的时程变化。在……里面 此外，他们将通过凝胶识别相关的蛋白质-DNA相互作用 移位和足迹分析，分离足迹蛋白并绘制 它们在发育过程中表达的时间和地点。他们还将 试图从整体上鉴定控制粘液细胞表型的基因 并鉴定激活其表达的间充质诱导因子。 在特定目标2下列出的研究将使用上皮细胞来确定 培养，MUC 2的内毒素上调是否先前观察到 在体内是直接作用或由中性粒细胞产物介导的。其他 研究将确定内毒素诱导粘蛋白的程度 转录或转录后和鉴定蛋白质-DNA 调节这种诱导的相互作用。因为最近的实验已经 发现假单胞菌(PA)产品上调MUC 2，附加 研究将通过类似的方法分析PA-MUC 2的诱导 用来分析内毒素-MUC-2的诱导并试图识别 PA上清液中的活性成分。从这些信息中获得的信息 研究将有望提出药物干预措施以抑制 人类呼吸道产生过多的粘蛋白。

Chronic obstructive airway diseases such as cystic fibrosis are characterized by mucin overproduction. Over time, this can lead to the formation of mucus plugs, infection, bronchiectasis, respiratory insufficiency, and death. An understanding of how this sequence of events is triggered would help in the design of methods to inhibit it. During the last period, our experiments confirmed the hypothesis that an early event in experimentally induced hypersecretion is the upregulation of steady state mucin (MUC 2) mRNA (induced by endotoxin instillation in rat airways). Our experiments also revealed a phasic upregulation of MUC 2 early in tracheal development. The goal of the proposed studies is to gain information about how these events are triggered and controlled. Studies listed under Specific Aim l will determine, using in situ hybridization, the time course of MUC 2 activation in the developing trachea. In addition, they will identify relevant protein-DNA interactions by gel shift and footprinting assays, isolate footprinted proteins and map the time and place of their expression during development. They will also attempt to identify genes controlling the mucous cell phenotype as a whole and identify mesenchymal induction factors activating their expression. Studies listed under Specific Aim 2 will determine, using epithelial cell cultures, whether the endotoxin upregulation of MUC 2 previously observed in vivo is a direct effect or is mediated by neutrophil products. Other studies will determine the extent to which mucin induction by endotoxin is transcriptional or post-transcriptional and identify protein-DNA interactions mediating this induction. Because recent experiments have revealed MUC 2 upregulation by Pseudomonas (PA) products, additional studies will analyze the PA-MUC 2 induction by methods similar to those used to analyze the endotoxin-MUC 2 induction and attempt to identify the active component in PA supernatant. The information gained from these studies will hopefully suggest pharmacological interventions to inhibit excessive mucin production in human airways.