Molecular Studies on MUC4 Mucin Gene

MUC4粘蛋白基因的分子研究

• 批准号: 7533778
• 负责人: Surinder K. Batra
• 金额: $ 22.4万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7533778
• 关键词: Actins; Adhesions; Adhesives; Alabama; Animal Model; Breeding; California; Cancer cell line; Carcinoma; Cell Polarity; Cell Proliferation; Cell Surface Proteins; Cell surface; Cells; Cloning; Co-Immunoprecipitations; Collaborations; Complementary DNA; Condition; Cytoplasm; Cytoplasmic Tail; Cytoskeleton; Data; Detection; Development; Diagnostic Neoplasm Staging; Disease; Disease Progression; Down-Regulation; Ductal Epithelial Cell; EGF-Like Domain; ERBB2 gene; Environment; Event; Exhibits; Exons; Extracellular Matrix; Fibroblasts; Funding; Genes; Genetic; Genomics; Glycoproteins; Growth; Human; In Vitro; Introns; Invasive; Knock-in Mouse; Laboratories; Length; Lesion; Localized; MUC4 mucin; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Membrane; Modeling; Molecular; Mucins; Mus; Mutation; Nature; Neoplasm Metastasis; Neoplasms; Nidogen; Normal Cell; Oncogenic; Other Genetics; Pancreas; Pancreatic Adenocarcinoma; Pancreatic Intraepithelial Neoplasia; Pathogenesis; Patients; Phenotype; Premalignant; Process; Property; Protein Overexpression; Proteins; Public Health; RNA Splicing; Receptor Protein-Tyrosine Kinases; Regulation; Reporting; Research; Research Personnel; Research Proposals; Role; San Francisco; Signal Transduction; Specificity; Staging; Structure; Study models; System; TP53 gene; Tandem Repeat Sequences; Testing; Thinking; Transgenic Mice; Transgenic Organisms; Tumorigenicity; Universities; Up-Regulation; Variant; Wild Type Mouse; Work; apomucin; base; cDNA Library; cancer cell; carcinogenesis; cell growth; cell motility; cell transformation; chronic pancreatitis; in vivo; insight; intraepithelial; mouse model; neoplastic; neoplastic cell; novel; outcome forecast; pancreatic neoplasm; programs; size; tumor growth; tumor progression; tumorigenic

DESCRIPTION (provided by applicant): The overall objective of this research proposal is to define the multifaceted role of the MUC4 mucin in the pathogenesis of pancreatic cancer (PC). During past funding cycles, we have made considerable progress, beginning with the cloning of the MUC4 cDNA from human pancreatic tumor cDNA libraries, establishing its complete genomic organization to understanding its function and regulation in PC cells. In our recent studies, we have shown a direct association of MUC4 with the tumorigenic and metastatic pancreatic cancer phenotype and provided experimental evidence for a functional role of MUC4 in growth and metastatic properties of PC cells. We have also demonstrated the oncogenic potential of MUC4 in NIH3T3 mouse fibroblast cells. Interestingly, our studies have also revealed that MUC4 regulates the expression of HER2 by post-transcriptional mechanism(s). We have shown that both MUC4 and HER2 physically interact with each other in PC cells. In this renewal application, we aim to establish the mechanistic basis for the pathological role of MUC4 in PC and to precisely define its cooperative action with other genetic aberrations during the course of pancreatic cancer pathogenesis. We hypothesize that the multi-domain structural features of MUC4 enable it to functionally participate in the early development and malignant process of PC cells via altering the normal cell-cell and cell-ECM (extra-cellular matrix) interactions, while simultaneously forming novel, cancer-specific interactions. To test our hypothesis, we propose three specific aims. In Aim 1, we will investigate the specific domain(s) or motif(s) in MUC4 responsible for its interaction with HER2, and determine the specificity and nature (direct or indirect) of interaction. In addition, we will define the mechanisms by which MUC4 regulates HER2 expression. Aim 2 will investigate the molecular mechanisms implicated in defining the multiple roles of MUC4 in tumor growth and metastasis. In Aim 3, we will investigate the cooperative action of MUC4 in combination with other defined oncogenic mutations during the early development of pancreatic cancer by generating MUC4-transgenic mice with conditional pancreas-specific expression. Additionally, we will investigate the cooperative action of MUC4 and K-rasG12D (oncogenic) in mouse models by cross-breeding. We will also utilize cultured mouse pancreatic ductal cells (PDCs), PDCs harboring K-ras mutations, and/or p53 inactivation to study MUC4 function(s) in syngeneic system. Taken together, these studies will establish the mechanistic basis for the role of MUC4 in early and late stages of pancreatic cancer progression. PUBLIC HEALTH RELEVANCE: The proposed second competitive research program in its 10th year aims to understand the mechanistic basis of the role of MUC4 in the pathogenesis of lethal pancreatic cancer (PC). During previous cycles, studies pertinent to MUC4 structure, function and regulation were performed in PC cells. MUC4 is aberrantly expressed in majority of pancreatic cancer cases compared to no detection in the normal pancreas. The proposed studies will investigate the precise role of MUC4 and its domains using human pancreatic cancer cell lines as well as in transgenic and syngeneic animal models.

描述（由申请人提供）：该研究建议的总体目的是定义MUC4粘蛋白在胰腺癌发病机理（PC）中的多方面作用。 在过去的融资周期中，我们取得了长足的进步，从人类胰腺肿瘤cDNA文库的MUC4 cDNA开始，建立了其完整的基因组组织，以了解其在PC细胞中的功能和调节。 在我们最近的研究中，我们显示了MUC4与肿瘤和转移性胰腺癌表型的直接关联，并为MUC4在PC细胞的生长和转移特性中的功能作用提供了实验证据。 我们还证明了MUC4在NIH3T3小鼠成纤维细胞中的致癌潜力。 有趣的是，我们的研究还表明，MUC4通过转录后机制调节HER2的表达。 我们已经表明，MUC4和HER2在PC细胞中彼此之间都有物理相互作用。 在此续签应用中，我们旨在为MUC4在PC中的病理作用建立机械基础，并在胰腺癌发病机理过程中精确定义其与其他遗传畸变的合作作用。 我们假设MUC4的多域结构特征使其能够通过改变正常的细胞细胞和细胞ECM（细胞外基质）相互作用，从而在PC细胞的早期发育和恶性过程中进行功能，同时形成新型的新型癌症，癌症特异性相互作用。 为了检验我们的假设，我们提出了三个具体目标。 在AIM 1中，我们将在MUC4中研究特定的域或基序，以使其与HER2相互作用，并确定相互作用的特异性和性质（直接或间接）。 此外，我们将定义MUC4调节HER2表达的机制。 AIM 2将研究与MUC4在肿瘤生长和转移中的多重作用有关的分子机制。 在AIM 3中，我们将通过在胰腺癌的早期发展过程中与其他定义的致癌突变结合使用MUC4的合作作用，通过产生具有条件胰腺特异性表达的MUC4-转基因小鼠。 此外，我们将通过跨繁殖在小鼠模型中研究MUC4和K-RASG12D（致癌）的合作作用。 我们还将利用培养的小鼠胰腺导管细胞（PDC），具有K-RAS突变的PDC和/或p53灭活来研究合成系统中的MUC4功能。 综上所述，这些研究将建立MUC4在胰腺癌进展的早期和晚期的作用的机械基础。 公共卫生相关性：拟议的第二年竞争研究计划旨在了解MUC4在致命胰腺癌发病机理（PC）中的作用的机理基础。 在先前的周期中，在PC细胞中进行了与MUC4结构，功能和调节有关的研究。 与正常胰腺未检测相比，大多数胰腺癌病例在大多数胰腺癌病例中都异常表达。 拟议的研究将使用人类胰腺癌细胞系以及转基因和同性动物模型研究MUC4及其结构域的精确作用。