Develop and Characterize a Novel Animal Model of Pancreatic Cancer

描述（由申请人提供）：忠实模拟人类疾病的胰腺导管腺癌（PDAC）动物模型对于研究人员开发有效的早期诊断、预防和治疗策略是必要的。近年来，已经开发了几种PDAC的遗传小鼠模型，并且极大地提高了我们对其生物学的理解。今天广泛使用的遗传模型是通过遗传改变胚胎胰腺或产生消化酶的成体细胞，胰腺腺泡细胞来开发的。这些模型忠实地模拟人类PDAC的发展是有争议的，因为人们普遍认为，在人类中，这种疾病在不同的细胞类型，导管细胞内发展。此外，PDAC是老年人的疾病，在儿童中没有发现。显然，基于成人胰管细胞遗传变化的遗传模型将为研究这种疾病提供重要的模型。先前开发依赖于导管特异性但非胰腺特异性的基因启动子（例如CK19）的胰腺导管特异性模型的尝试已经失败，因为癌基因在所有CK19阳性细胞中的广泛表达是胚胎致死的。在拟议的项目中，我们将开发一种小鼠模型，其中基因表达在成年胰腺导管细胞而不是任何其他细胞中受到特异性控制。为了做到这一点，我们开发了一种新的策略，使用两个必须串联激活的基因启动子的组合。对于导管细胞特异性，我们将使用充分表征的CK19启动子，其在整个身体的导管细胞中具有活性。对于胰腺特异性，我们将使用PDX1启动子，其在胚胎发育期间在胰腺前体细胞中表达。我们将这些启动子组装在一个构建体中，使得导管细胞特异性启动子CK19在胰腺特异性启动子PDX1的控制下。为了暂时控制基因表达，我们将利用我们的系统来表达一种可以被药物他莫昔芬激活的分子。一旦被诱导，这种分子就能够激活或删除专门为此目的而设计的小鼠中的基因，其中许多基因已经可用。该模型将首次允许胰管细胞基因表达的特异性调节，这对于许多生理和病理研究是有用的。一旦胰管特异性表达系统建立，我们将用它来激活癌基因并抑制肿瘤抑制因子，以模拟人类中常见的情况来产生PDAC。在该模型中产生的PDAC样本将与来自其他遗传小鼠模型和人类疾病的样本进行比较。我们期望这种胰腺癌动物模型将更具有临床相关性，并适合于转化研究。

DESCRIPTION (provided by applicant): Animal models of pancreatic ductal adenocarcinoma (PDAC) that faithfully mimic the human disease are necessary to allow researchers to develop effective early diagnosis, prevention, and treatment strategies. In recent years, several genetic mouse models of PDAC have been developed and have greatly improved our understanding of its biology. The genetic models in widespread usage today were developed by genetically altering either the embryonic pancreas or the adult cells that produce digestive enzymes, pancreatic acinar cells. That these models faithfully mimic the development of human PDAC is controversial because it is generally felt that in humans this disease develops within a different cell type, duct cells. Furthermore, PDAC is a disease of the elderly and is not found in children. Clearly, a genetic model based on genetical changes in adult pancreatic duct cells would provide an important model for investigation of this disease. Previous attempts to develop pancreatic duct-specific models relying on gene promoters that were duct specific but not pancreas specific (e.g. CK19) have failed because broad expression of oncogenes in all CK19-positive cells is embryonically lethal. In the proposed project, we will develop a mouse model in which gene expression is controlled specifically in adult pancreatic duct cells rather than any other cells. In order to do so, we have developed a novel strategy that uses a combination of two gene promoters that must be activated in tandem. For duct cell specificity, we will use the well-characterized CK19 promoter, which is active in duct cells throughout the body. For pancreatic specificity, we will use the PDX1 promoter, which is expressed in pancreatic precursor cells during embryonic development. We will assemble these promoters in a construct such that the duct cell- specific promoter CK19 is under the control of the pancreas-specific promoter PDX1. To control gene expression temporally, we will utilize our system to express a molecule that can be activated by a drug, tamoxifen. Once induced, this molecular is capable of activating or deleting genes in mice engineered specifically for this purpose, many of which are already available. This model will allow for the first time the specific regulation of pancreatic duct cell gene expression, which is useful for many physiological and pathological studies. Once the pancreatic duct-specific expression system is established, we will use it to activate an oncogene and inhibit a tumor suppressor to mimic the situation commonly found in humans to generate PDAC. Specimens of the PDAC that develops in this model will be compared with those from other genetic mouse models and with the human disease. We expect that this animal model of pancreatic cancer will be more clinically relevant and appropriate for translational research.