EGFRvIII in Pancreatic Cancer

胰腺癌中的 EGFRvIII

• 批准号: 7874684
• 负责人: MANEESH JAIN
• 金额: $ 7.43万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-19 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7874684
• 关键词: Agar; Alternative Splicing; Amino Acids; Antibodies; Appearance; Applications Grants; Behavior; Biological Markers; Breast; Cancer Etiology; Cancer cell line; Cells; Cessation of life; Cysteine; DNA Sequence Rearrangement; Data; Development; Diagnosis; Diagnostic; Drug resistance; Ductal Epithelial Cell; EGFR Gene Amplification; EGFR Protein Overexpression; Early Diagnosis; Epidermal Growth Factor Receptor; Epithelial; Epitopes; Exons; Fibroblasts; Genes; Glioblastoma; Glycine; Goals; Head and Neck Cancer; Head and neck structure; Human; In Vitro; Incidence; Incidence Study; Investigation; Lead; Ligands; Lung; Malignant Neoplasms; Malignant neoplasm of pancreas; Malignant neoplasm of prostate; Messenger RNA; Monoclonal Antibodies; Mus; Neoplastic Cell Transformation; Non-Small-Cell Lung Carcinoma; Nude Mice; Oncogenic; Ovarian; Pancreas; Pancreatic Adenocarcinoma; Pathogenesis; Pharmaceutical Preparations; Phenotype; Play; Principal Investigator; Prostate; Proteins; RNA; Radioimmunotherapy; Radioisotopes; Relapse; Reporting; Research; Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Rodent; Role; Sampling; Signal Pathway; Solid Neoplasm; Testing; Therapeutic; Therapeutic Intervention; Tissues; Toxin; Training; Tumor Antigens; Tumor Markers; Tumor Tissue; United States; Variant; Virus; base; cancer cell; cancer type; extracellular; in vivo; mRNA Expression; malignant phenotype; metastatic process; mutant; neoplastic cell; novel; novel marker; outcome forecast; overexpression; pancreatic neoplasm; public health relevance; receptor; tumor; tumorigenic

DESCRIPTION (provided by applicant): Pancreatic cancer is the fourth leading cause of cancer related death in the United States. There is an urgent need to identify novel biomarkers for early diagnosis and targets for therapeutic interventions. Overexpression of EGFR in PC cells plays a crucial role in the uncontrolled proliferation of neoplastic cells. Amplification or overexpression of the wild-type EGFR (EGFRwt) gene is frequently associated with rearrangements or alternate splicing resulting in the expression of structurally altered EGFR mRNA and protein. EGFRvIII is the most is the most common naturally occurring mutant form of EGFR and is expressed in many cancer types including glioblastoma, breast, ovarian, head and neck, lung and prostate cancer. Although amplification of EGFR and its ligands is well documented in pancreatic cancer, no efforts have been made to study the incidence or involvement of EGFRvIII in pancreatic cancer. Our preliminary results suggest that EGFRvIII is highly expressed in pancreatic cancer tissues. Based on the preliminary findings we hypothesize that "naturally occurring epidermal growth factor receptor variant III (EGFRvIII) contributes to the pathogenesis of pancreatic adenocarcinomas". To test the hypothesis following specific aims are proposed: SPECIFIC AIM I will investigate the expression of EGFR and EGFR variants in human pancreatic tumors by reverse transcriptase- polymerase chain reaction (RT-PCR) and immunohistochemical analysis. PCR- amplification will demonstrate the expression of mRNA. Immunohistochemical analysis using a EGFRvIII specific monoclonal antibody will indicate the presence and cellular localization of the EGFRvIII protein. This aim will provide the information on the incidence of expression of EGFRvIII in human pancreatic tumors. SPECIFIC AIM II will express EGFRvIII in highly tumorigenic (HPAF) and poorly tumorigenic (MiaPaCa) human pancreatic cancer cell lines and immortalized human pancreatic ductal cells (HTERT/HPNE) to establish its contribution to the malignant phenotype. This aim will support our prediction that expression of EGFRvIII in cultured immortal pancreatic cells is sufficient to lead to a transformed phenotype; that the cells will gain the ability to form foci in soft agar and to form tumors in nude mice. Taken together, these studies will provide the incidence and function of a novel naturally occurring variant of EGFRvIII in the lethal pancreatic cancer. The long-term goal of this project is to use the newly developed anti-EGFRvIII antibodies for targeted therapy of pancreatic cancer. PUBLIC HEALTH RELEVANCE: Mutant epidermal growth factor receptor, EGFRvIII is an attractive target for targeted therapy due to its tumor-restricted expression and presence of unique antigenic epitopes. In this grant application, we propose to investigate EGFRvIII's expression in pancreatic tumors (various grades of malignancy), its oncogenic function in immortal pancreatic cells, and its effect on tumorigenecity of pancreatic cancer cells.

描述(申请人提供)：胰腺癌是美国癌症相关死亡的第四大原因。迫切需要为早期诊断和治疗干预确定新的生物标记物。EGFR在PC细胞中的过表达在肿瘤细胞的失控增殖中起着重要作用。野生型EGFR基因的扩增或过表达经常与重排或交替剪接有关，导致结构改变的EGFR mRNA和蛋白的表达。EGFRvIII是最常见的自然发生的EGFR突变形式，在许多癌症类型中都有表达，包括胶质母细胞瘤、乳腺癌、卵巢癌、头颈部癌、肺癌和前列腺癌。尽管EGFR及其配体在胰腺癌中的扩增已有很好的文献报道，但尚未有人致力于研究EGFRvIII在胰腺癌中的发生率或参与。我们的初步结果表明，EGFRvIII在胰腺癌组织中高表达。根据初步发现，我们假设“自然发生的表皮生长因子受体变异体III(EGFRvIII)与胰腺癌的发病机制有关”。为了验证这一假设，提出了以下具体目标：特异性目的I将通过逆转录聚合酶链式反应(RT-PCR)和免疫组织化学分析来研究EGFR及其变异体在人胰腺肿瘤中的表达。用聚合酶链式反应(PCR)扩增后可检测到mRNA的表达。使用EGFRvIII特异性单抗的免疫组织化学分析将表明EGFRvIII蛋白的存在和细胞定位。这将为研究EGFRvIII在人胰腺肿瘤中的表达情况提供信息。特异性AIM II将在高致瘤性(HPAF)和低致瘤性(MiaPaCa)人胰腺癌细胞系和永生化人胰腺导管细胞(hTERT/HPNE)中表达EGFRvIII，以确定其在恶性表型中的作用。这一目标将支持我们的预测，即在培养的永生胰腺细胞中表达EGFRvIII足以导致表型转化；该细胞将获得在软琼脂中形成病灶的能力，并在裸鼠体内形成肿瘤。综上所述，这些研究将提供一种新的自然发生的EGFRvIII变异在致命性胰腺癌中的发生率和功能。该项目的长期目标是将新开发的抗EGFRvIII抗体用于胰腺癌的靶向治疗。公共卫生相关性：突变的表皮生长因子受体，EGFRvIII，由于其肿瘤限制性表达和独特的抗原表位的存在，是有吸引力的靶向治疗的目标。在这项拨款申请中，我们建议研究EGFRvIII在胰腺肿瘤(不同级别的恶性肿瘤)中的表达，它在永生胰腺细胞中的致癌功能，以及它对胰腺癌细胞肿瘤发生的影响。