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（由申请人提供）：胰腺癌是美国第四大癌症死亡原因，5% 的五年生存率表明需要改进治疗方法，更好地了解其机制。可能决定肿瘤表型的几个因素包括：1）肿瘤来源的细胞类型，2）转化细胞获得的聚集基因突变，以及3）获得基因突变的序列基于先前的研究和我们的初步研究。数据，我的假设是，细胞起源是决定胰腺肿瘤表型的重要因素，尽管胰腺导管腺癌（PDA）是最常见的胰腺癌形式，但我们仍然不知道是哪种细胞类型引起的。对于其他类型的胰腺肿瘤也是如此。我的目标是研究成人胰腺腺泡细胞 (AC) 或导管细胞 (DC) 特异表达的突变的致瘤作用。我将使用两种小鼠模型。通过 k-ras 和 Trp53 的并发突变复制 PDA；第二个是我们实验室最近开发的基于外源激活的实体假乳头状瘤 (SPT) 的新模型； -catenin 通过 Cre 重组-外显子 3 (¿-catex3) 的切除 我的第一个目标是仅针对成年小鼠中的 AC 进行突变，我将杂交携带突变 k-ras 和 Trp53 (k-rasG12D/) 条件表达等位基因的小鼠。 Trp53R172H) 或 ¿ -catenin (¿-catex3) 到携带他莫昔芬激活的 Cre 的小鼠中，这些小鼠通过弹性蛋白酶基因衍生的启动子表达到 AC，将通过定量 PCR 来分析肿瘤的组织学和免疫组织化学特征。是评估 k-rasG12D/Trp53R172H 或 ¿ 的效果DC中的catex3。目前，没有启动子将Cre仅引导至导管上皮，相反，我将从成人krasG12D/Trp53R172H或¿中分离和培养DC。 -catex3 小鼠，并同时引入慢病毒递送的 Cre 体外激活它们，以便通过发光体内监测肿瘤生长，我们将这些细胞重新植入裸鼠中，并以与 Aim 中相同的方式表征形成的肿瘤。 1. 公共健康相关性：识别引起胰腺肿瘤的细胞类型具有几个重要意义：首先，我们将能够将更多的研究工作集中在特定的细胞类型上，并对其进行更详细的研究。其次，我们也许能够提供更有针对性的治疗，提高疗效并减少副作用。