Defining the Contributions of Pancreatic Ductal and Acinar Cells to Tumorigenesis

定义胰管和腺泡细胞对肿瘤发生的贡献

• 批准号: 7686161
• 负责人: Sam C. Wang
• 金额: $ 5.34万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-03 至 2010-07-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7686161
• 关键词: Acinar Cell; Address; Adenocarcinoma; Adopted; Adult; Adverse effects; Alleles; Bypass; Cancer Etiology; Cells; Cessation of life; Characteristics; Conflict (Psychology); Cultured Cells; Data; Development; Disease; Disease Progression; Duct (organ) structure; Ductal; Ductal Epithelial Cell; Ductal Epithelium; Elastases; Evaluation; Excision; Exons; Gene Expression; Gene Mutation; Genes; Genetic; Genetic Recombination; Goals; Growth; Histologic; Histology; Human; Hyperplasia; Immunohistochemistry; Implant; In Situ; In Vitro; Indolent; Injection of therapeutic agent; Intraepithelial Neoplasia; Islet Cell; Laboratories; Lead; Lesion; Luciferases; Malignant Neoplasms; Malignant neoplasm of pancreas; Metaplasia; Mixed Neoplasm; Modeling; Molecular Profiling; Monitor; Morphology; Mus; Mutate; Mutation; Names; Neoplasm Metastasis; Newly Diagnosed; Nude Mice; Orthologous Gene; Pancreas; Pancreatic Ductal Adenocarcinoma; Pancreatic Injury; Pancreatitis; Phase; Phenotype; Play; Population; Research; Role; Sampling; Shapes; Signal Transduction; Solid; Solid Neoplasm; Source; Subfamily lentivirinae; Surface Antigens; Survival Rate; TP53 gene; Tamoxifen; Testing; Time; Transgenic Mice; Tubular formation; Tumor Suppressor Proteins; United States; base; blastomere structure; carcinogenesis; cell transformation; cell type; chronic pancreatitis; defined contribution; human disease; improved; in vivo; insight; intraperitoneal; luminescence; mouse model; novel; pancreatic neoplasm; pancreatic tumorigenesis; promoter; public health relevance; recombinase; response; tool; tumor; tumor growth; tumorigenesis; tumorigenic

DESCRIPTION (provided by applicant): Pancreatic cancer is the fourth-leading cause of cancer deaths in the United States. The 5% five-year survival rate demonstrates the need for improved treatments. To address that need, a better understanding of the mechanisms of pancreatic carcinogenesis is needed. Several factors that may determine tumor phenotype include: 1) cell type of tumor origin, 2) aggregate genetic mutations acquired by transformed cells, and 3) sequence by which genetic mutations are acquired. Based on previous studies and our preliminary data, my hypothesis is that cellular origin is an important factor in determining pancreatic tumor phenotype. Although pancreatic ductal adenocarcinoma (PDA) is the most common form of pancreatic cancer, we still do not know which cell type gives rise to it. This lack of insight is also true for other types of pancreatic tumors. My objectives are to study the tumorigenic effects of mutations expressed specifically to adult pancreatic acinar cells (AC) or ductal cells (DC). I will use two mouse models. The first reproduces PDA via concurrent mutations in k-ras and Trp53; the second is a novel model for solid pseudopapillary tumor (SPT) that our laboratory recently developed and is based on exogenously activated ¿-catenin via Cre recombination-excision of exon 3 (¿-catex3). My first aim is to target mutations to only AC in adult mice. I will cross mice carrying conditionally expressed alleles of mutated k-ras and Trp53 (k-rasG12D/Trp53R172H) or ¿-catenin (¿-catex3) to mice carrying tamoxifen-activated Cre that are expressed to AC via a promoter derived from the elastase gene. Tumors will be characterized with histology and immunohistochemistry. Gene expression profiles will be analyzed by quantitative PCR. My second aim is to evaluate the effects of k-rasG12D/Trp53R172H or ¿-catex3 in DC. Currently, there is no promoter to direct Cre to only ductal epithelium. Instead, I will isolate and culture DC from adult krasG12D/Trp53R172H or ¿-catex3 mice and activate them in vitro with lentiviral delivered Cre. Luciferase will be concurrently introduced to allow in vivo monitoring of tumor growth via luminescence. We will re-implant these cells into nude mice and characterize formed tumors in the same manner as in Aim 1. PUBLIC HEALTH RELEVANCE: Identifying the types of cells that give rise to pancreatic tumors has several important implications. First, we will be able to focus additional research efforts towards that particular cell type and pursue more detailed studies into the mechanisms behind pancreatic tumorigenesis. Secondly, we may be able to deliver more targeted therapy that has increased efficacy and reduced side effects.

描述（由适用提供）：胰腺癌是美国癌症死亡的第四个主要原因。 5％的五年生存率表明需要改善治疗方法。为了满足需求，需要更好地了解胰腺癌的机制。可能决定肿瘤表型的几个因素包括：1）肿瘤起源的细胞类型，2）通过转化细胞获得的聚集遗传突变，以及3）序列获得了遗传突变。根据先前的研究和我们的初步数据，我的假设是细胞起源是确定胰腺肿瘤表型的重要因素。尽管胰腺导管腺癌（PDA）是胰腺癌最常见的形式，但我们仍然不知道哪种细胞类型会引起它。缺乏洞察力对于其他类型的胰腺肿瘤也是如此。我的目标是研究针对成年胰腺腺泡细胞（AC）或导管细胞（DC）的突变的致瘤作用。我将使用两种鼠标型号。第一个通过K-RAS和TRP53中的并发突变复制PDA；第二个是我们实验室最近开发的固体假毛细血管肿瘤（SPT）的新型模型，并基于外源激活的 - 通过外显子3（€-CATEX3）的CRE重组 - 分解 - catenin。我的第一个目的是将突变靶向仅在成年小鼠中。我将横穿携带有条件表达的K-RAS和TRP53（K-Rasg12d/Trp53r172H）或�-catenin（�-Catex3）的小鼠，向携带他莫昔芬激活的CRE的小鼠，这些Cre通过启动子通过弹性蛋白衍生而来的AC表达为AC。肿瘤将以组织学和免疫组织化学为特征。基因表达谱将通过定量PCR分析。我的第二个目的是评估DC中K -Rasg12d/trp53r172h或� -catex3的效果。当前，尚无启动子将CRE引导到仅导管上皮。取而代之的是，我将与成年Krasg12d/trp53r172h或� -catex3小鼠隔离和培养DC，并用慢跑病毒递送的CRE在体外激活它们。将同时引入荧光素酶，以允许通过发光对肿瘤生长进行体内监测。我们将将这些细胞重新植入裸鼠中，并以与AIM 1相同的方式表征形成的肿瘤。首先，我们将能够将更多的研究工作集中在该特定的细胞类型上，并将更详细的研究购买用于胰腺肿瘤发生的机制。其次，我们也许能够提供更有针对性的治疗，从而提高了效率并降低了副作用。