Biomarker Discovery for Early Detection of Pancreatic Ductal Adenocarcinoma

项目总结（见说明）： 浸润性胰腺导管腺癌（PDAC）是人类最致命的恶性肿瘤之一， 五年存活率不到百分之五与PDAC相关的高死亡率是 这主要是由于疾病诊断的晚期和胰腺癌对化疗的抗性。 传统的化疗和放疗治疗方法。这些观察突出了关键的 需要在手术切除可以治愈的阶段早期发现胰腺肿瘤。的 这项提案的主要目标是使用最先进的质谱分析和基因工程 小鼠模型代表胰腺肿瘤的浸润前或早期，器官局限阶段，以确定 一套全面的生物标志物，可联合用于前瞻性诊断早期肿瘤 人类的疾病。在目标1中，我们将在表达一种免疫缺陷蛋白的遗传近交系小鼠中鉴定血清生物标志物。 胰腺导管上皮和KrasG 12 D小鼠中Kras的激活等位基因（KrasG 12 D） 在p16 lnk 4a或p53缺陷背景下表达。KRAS的组成性激活在超过 90%的PDAC病例，它可能代表胰腺癌中最早的遗传病变之一。目的2 将利用来自目标1中所述的相同动物模型的新衍生的胰腺导管细胞系， 表征细胞表面蛋白质组和分泌组中发生的变化的完整谱 随着肿瘤从浸润前病变进展到完全转移性疾病。在目标3中，我们将利用 组织学引导MALDI-TOF MS组织分析以鉴定与早期 使用来自动物模型的胰腺组织的胰腺瘤形成。根据这些结果，我们预计 编制一套全面的生物标志物，用于检测早期胰腺癌。最后，在目标4中， 我们将测试在小鼠系统中鉴定的早期生物标志物的特异性和灵敏度， 使用从健康供体收集的人血清样品诊断胰腺肿瘤的能力， 代表慢性胰腺炎和PDAC的病例。这些研究的目标将是发展一种 基于抗体的诊断分析，可用于筛查胰腺癌的高危患者。

PROJECT SUMMARY (See instructions): Infiltrating pancreatic ductal adenocarcinoma (PDAC) is one of the most fatal human malignancies, with an overall five-year survival rate of less than five percent. The high mortality rate associated w/ith PDAC is primarily due to the late stage at which disease is diagnosed and the resistance of pancreatic cancer to traditional chemotherapy and radiotherapy treatment approaches. These observations highlight the critical need for early detection of pancreatic neoplasia at a stage where surgical resection can be curative. The major goal of this proposal will be to use state-of-the-art mass spectrometry and genetically engineered mouse models representing preinvasive or early, organ-confined stages of pancreatic neoplasia to identify a comprehensive set of biomarkers that can be used in combination to prospectively diagnose early neoplasic disease in humans. In Aim 1, we will identify serum biomarkers in genetically inbred mice that express an activated allele of Kras (KrasG12D) in the pancreatic ductal epithelium and in mice with KrasG12D expressed on a p16lnk4a- or p53-deficient background. Constitutive activation of KRAS is observed in over 90% of PDAC cases and it likely represents one of the earliest genetic lesions in pancreatic cancer. Aim 2 will utilize newly-derived pancreatic ductal cell lines from the same animal models described in Aim 1 to characterize the complete spectrum of changes that occur in the cell-surface proteome and in the secretome as neoplasia progresses from preinvasive lesions to fully metastatic disease. In Aim 3, we will utilize histology-guided MALDI-TOF MS tissue profiling to identify biomarkers associated with early-stage pancreatic neoplasia using pancreatic tissue from the animal models. From these results, we expect to compile a comprehensive set of biomarkers for detection of early-stage pancreatic cancer. Finally, in Aim 4, we will test the specificity and sensitivity of the early-stage biomarkers identified in the murine system for their ability to diagnose pancreatic neoplasia using human serum samples collected from healthy donors and cases representing chronic pancreatitis and PDAC. The goal of these studies will be the development of an antibody-based diagnostic assay that can be used to screen high-risk patients for pancreatic cancer.