TRANSGENIC MOUSE MODELS OF PANCREATIC EXOCRINE FUNCTION

胰腺外分泌功能的转基因小鼠模型

• 批准号: 6416519
• 负责人: ROBERT C DE LISLE
• 金额: $ 15万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-06-01 至 2004-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6416519
• 关键词: acinar cell; disease /disorder model; exocytosis; gene targeting; genetically modified animals; glycoprotein structure; granule; laboratory mouse; model design /development; pancreas; pancreatitis; zymogens

DESCRIPTION (provided by applicant): This grant application proposes to develop genetically engineered mouse models for acute pancreatitis and cystic fibrosis-related pancreatitis. We will focus on a recently discovered gene, which preliminary data suggest may play a role in development of pancreatitis, and also in the pathogenesis of cystic fibrosis (CF). The gene is called crpd in the mouse and dmbt1 in humans and it encodes the protein Muclin, a high molecular weight, and sulfated glycoprotein. This gene is most highly expressed in the exocrine pancreas, but also as different transcripts in other organs including the intestine, gallbladder, and kidney. The hypothesis is that Muclin is required for normal zymogen granule formation and subsequent solubilization of digestive enzymes upon exocytosis of the granules. We predict that if this gene is a disrupted zymogen granule will fail to mature creating a situation in the acinar cell where the zymogens will be prone to become activated, resulting in pancreatitis. The Muclin-deficient mouse model is intended to explore this hypothesis and, if this is borne out, it will make looking for mutations in this gene in human idiopathic pancreatitis a reasonable endeavor. We also predict that mice over expressing Muclin will exhibit poor release of zymogens after exocytosis and will be prone to CF-like plugging of the acinar lumen, which may predispose to pancreatitis. In CFTR-/- mice, a model for CF, the pancreas exhibits over expression of Muclin, which is associated with protein, plugs in the acinar lumen. Mice with a Muclin transgene targeted to the pancreas will allow testing of this hypothesis. The specific aims are to produce genetically engineered mice: 1). Globally deficient in Muclin by targeting the first exon, which is present in all transcripts of this gene. 2). An exocrine cell-specific defective Muclin by targeting the last exon which is translated only in exocrine cell transcripts of the gene and codes for a transmembrane domain that may be important for Muclin' s exocrine function; 3). That over expresses Muclin in the pancreatic acinar cell using the rat elastase promoter to direct cell specific expression.

描述（申请人提供）：本赠款申请提议开发 用于急性胰腺炎和囊性的基因工程小鼠模型 与纤维化有关的胰腺炎。我们将专注于最近发现的基因 哪些初步数据表明可能在胰腺炎的发展中起作用， 以及囊性纤维化（CF）的发病机理。该基因称为CRPD 在人类中的小鼠和DMBT1中，它编码蛋白质muclin 分子量和硫酸糖蛋白。该基因表达最高 在外分泌胰腺中，但在其他器官中也是不同的成绩单 包括肠道，胆囊和肾脏。假设是穆克林 是正常的扎原颗粒形成和随后的溶解所必需的 颗粒胞吐作用后的消化酶。我们预测如果这个 基因是一个破坏的酶原颗粒，将无法成熟 酶细胞将容易被激活，因此 在胰腺炎中。 Muclin缺陷鼠标模型旨在探讨这一点 假设，如果证明这一点，它将成为寻找突变 人类特发性胰腺炎中的该基因是合理的努力。我们也是 预测超过表达穆克林的小鼠将表现出Zymogens的不良释放 胞吞作用后，将容易被刺激性腔内插入的CF状插入， 这可能易于胰腺炎。在CFTR - / - 小鼠中，CF的模型 胰腺表现出与蛋白质相关的Muclin的表达 插入腺泡腔。具有粘液转基因的小鼠针对 胰腺将允许检验该假设。具体目的是 产生基因工程的小鼠：1）。全球缺乏穆克林 靶向第一个外显子，该外显子存在于该基因的所有转录本中。 2）。 通过靶向最后一个外显子的外分泌细胞特异性有缺陷的muclin 仅在基因的外分泌细胞转录本中翻译并编码 跨膜结构域可能对Muclin的外分泌功能很重要； 3）。 使用大鼠弹性酶，超过胰腺腺泡细胞中的muclin 启动子直接特定细胞表达。