LCM & 2D PAGE in Early Detection of Pancreatic Cancer

描述（由申请人提供）： 本申请的目的是鉴定胰腺癌患者血清特异性识别的胰腺癌细胞特异性抗原，并在申请人的实验室中建立研究胰腺癌的新领域。这项研究的长期目标是开发有效的方法来检测，筛查，预防和治疗人类胰腺癌。每年，在这个国家发现30，300例新的胰腺癌病例。可悲的是，由于诊断困难，疾病的内在侵略性以及有效的系统治疗的不可用性，每年死于该疾病的人数大约相同。许多遗传分析研究和基于蛋白质组学的方法已被用于鉴定胰腺癌中可用作生物标志物的改变的基因、蛋白质或抗原。不幸的是，使用来自胰腺癌患者的整个活检标本或细胞培养物导致对胰腺癌具有低特异性的标志物的鉴定。我们的实验室最近应用新技术分离纯癌细胞，并鉴定出与胰腺癌患者血清反应而不与健康志愿者血清反应的胰腺癌抗原。基于这些结果，我们假设可以通过筛选胰腺癌细胞特异性蛋白与胰腺癌患者血清中自身抗体的反应性来鉴定肿瘤细胞特异性抗原。为了测试这一点，我们最近使用了最先进的技术，如激光捕获显微切割（LCM），二维凝胶电泳（2D-PAGE），和蛋白质印迹分析，以确定肿瘤特异性抗原。在这里，我们提出鉴定胰腺癌细胞特异性抗原，其仅与来自胰腺癌患者的血清反应，而不与来自其他胰腺疾病患者、患有其他恶性肿瘤（例如GI和乳腺癌）的患者的血清和来自健康个体的血清反应。我们的具体目标是：i）鉴定与来自胰腺癌患者的血清中的自身抗体反应的癌细胞特异性抗原，ii）确定抗原对胰腺癌的特异性，iii）确定胰腺癌细胞特异性抗原的身份。这些癌细胞特异性抗原可作为胰腺癌早期检测、筛查和靶向治疗的生物标志物。

DESCRIPTION (provided by applicant): The goal of this application is to identify pancreatic cancer cell-specific antigens that are specifically recognized by sera from patients with pancreatic carcinoma and to establish a new area of research in the applicant's laboratory to investigate pancreatic cancer. The long-term goal of this research is to develop effective means to detect, screen, prevent, and treat pancreatic cancer in humans. Each year, 30,300 new cases of pancreatic cancer are detected in this country. Sadly, about the same number die per year from the disease due to difficulties in diagnosis, the intrinsic aggressive nature of the disease, and the unavailability of effective systemic treatment. Many genetic profiling studies and proteomic-based approaches have been used to identify altered genes, proteins or antigens in pancreatic cancer that could be used as biomarkers. Unfortunately, the use of entire biopsy specimens or cell cultures from pancreatic cancer patients led to identification of markers with low specificity for pancreatic cancer. Our laboratory recently applied new technologies in isolating pure cancer cells and identified pancreatic cancer antigens that reacted with sera from patients with pancreatic cancer and not with sera from healthy volunteers. Based on these results, we hypothesized that tumor cell-specific antigen(s) can be identified by screening pancreatic cancer cell-specific proteins for reactivity with autoantibodies from sera of pancreatic carcinoma patients. To test this, we recently used state-of-the-art technologies such laser capture microdissection (LCM), 2-dimensional gel electrophoresis (2D-PAGE), and Western blot analysis to identify tumor specific antigens. Here, we propose to identify pancreatic cancer cell-specific antigens that will react only with sera from patients with pancreatic cancer and not with sera from patients with other pancreatic diseases, patients with other malignancies (e.g. Gl and breast), and sera from healthy individuals. Our specific aims are: i) to identify carcinoma cell-specific antigen(s) that react with autoantibodies in sera from patients with pancreatic carcinoma, ii) to determine the specificity of the antigen(s) for pancreatic cancer, iii) to determine the identity of the pancreatic cancer cell-specific antigen(s). These cancer cell-specific antigens could be developed as biomarkers for early detection, screening, and targeted therapy of pancreatic cancer.