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: 6620381
• 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 2005-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6620381
• 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（由申请人提供）：本赠款申请建议开发 急性胰腺炎和囊性胰腺炎的基因工程小鼠模型 纤维化相关胰腺炎我们将关注最近发现的一个基因， 初步数据表明其可能在胰腺炎的发展中起作用， 以及囊性纤维化（CF）的发病机制。该基因被称为crpd 在小鼠和人类dmbt 1，它编码的蛋白质Muclin，一个高 分子量和硫酸化糖蛋白。这个基因在人类的大脑中 在胰腺外分泌中，也在其他器官中作为不同的转录物 包括肠胆囊和肾假设穆克林 是正常酶原颗粒形成和随后溶解所必需的 消化酶的分泌。我们预测，如果这 基因被破坏的酶原颗粒将无法成熟， 腺泡细胞中的酶原将容易被激活， 胰腺炎Muclin缺陷小鼠模型旨在探索这一点 假设，如果这一点得到证实，它将使寻找突变， 这个基因在人类特发性胰腺炎中是一个合理的奋进。我们也 预测过表达Muclin的小鼠将表现出酶原的不良释放 在胞吐作用之后，并且将易于发生腺泡腔的CF样堵塞， 这可能导致胰腺炎。在CFTR-/-小鼠中，CF模型， 胰腺表现出与蛋白质相关的Muclin的过度表达， 堵塞腺泡腔带有Muclin转基因的小鼠靶向 胰腺可以检验这个假设。具体目标是 产生基因工程小鼠：1）。全球缺乏Muclin， 靶向存在于该基因的所有转录物中的第一外显子。2）。 通过靶向最后一个外显子， 只在外分泌细胞中翻译的基因转录本， 可能对Muclin的外分泌功能重要的跨膜结构域; 3）。 利用大鼠弹性蛋白酶在胰腺腺泡细胞中过度表达Muclin 启动子以指导细胞特异性表达。