EGFRvIII in Pancreatic Cancer

胰腺癌中的 EGFRvIII

• 批准号: 7641980
• 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 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7641980
• 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（由申请人提供）：胰腺癌是美国癌症相关死亡的第四大原因。迫切需要鉴定用于早期诊断的新型生物标志物和用于治疗干预的靶点。EGFR在PC细胞中的过表达在肿瘤细胞的不受控制的增殖中起着至关重要的作用。野生型EGFR（EGFRwt）基因的扩增或过表达通常与重排或选择性剪接相关，导致结构改变的EGFR mRNA和蛋白质的表达。EGFRvIII是EGFR最常见的天然突变形式，在许多癌症类型中表达，包括胶质母细胞瘤、乳腺癌、卵巢癌、头颈癌、肺癌和前列腺癌。尽管EGFR及其配体的扩增在胰腺癌中有充分的记载，但尚未努力研究胰腺癌中EGFRvIII的发病率或参与。我们的初步结果表明，EGFRvIII是高表达的胰腺癌组织。基于初步的研究结果，我们假设“天然存在的表皮生长因子受体变体III（EGFRvIII）有助于胰腺癌的发病机制”。为了验证这一假设，提出了以下具体目标：具体目的我将通过逆转录-聚合酶链反应（RT-PCR）和免疫组化分析研究EGFR及其变体在人胰腺肿瘤中的表达。PCR扩增将证明mRNA的表达。使用EGFRvIII特异性单克隆抗体的免疫组织化学分析将指示EGFRvIII蛋白的存在和细胞定位。这一目标将提供有关EGFRvIII在人类胰腺肿瘤中表达发生率的信息。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，以确定其对恶性表型的贡献。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足以导致转化的表型;细胞将获得在软琼脂中形成病灶和在裸鼠中形成肿瘤的能力。总之，这些研究将提供一种新的天然存在的EGFRvIII变异体在致命性胰腺癌中的发病率和功能。本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。公共卫生关系：突变型表皮生长因子受体（EGFRvIII）由于其肿瘤限制性表达和存在独特的抗原表位而成为靶向治疗的有吸引力的靶点。在本申请中，我们计划研究EGFRvIII在胰腺肿瘤（各种恶性程度）中的表达，其在永生胰腺细胞中的致癌功能，以及其对胰腺癌细胞致瘤性的影响。