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，具有 总体五年存活率不到5%。与PDAC相关的高死亡率是 主要是由于疾病诊断的晚期以及胰腺癌对 传统的化疗和放射治疗方法。这些观察结果突显了 需要在手术切除可以治愈的阶段及早发现胰腺肿瘤。这个 这项提案的主要目标将是使用最先进的质谱学和基因工程 代表胰腺肿瘤侵袭前或早期器官受限阶段的小鼠模型 一套全面的生物标志物，可以结合使用来前瞻性地诊断早期肿瘤 人类的疾病。在目标1中，我们将在基因近交系小鼠中鉴定表达An的血清生物标记物。 Kras(KrasG12D)在胰腺导管上皮和KrasG12D小鼠中的激活等位基因 在p16lnk4a或p53缺失的背景下表达。观察到了KRAS的结构性激活。 90%的PDAC病例，它可能是胰腺癌最早的遗传损害之一。目标2 将利用来自目标1中描述的相同动物模型的新来源的胰腺导管细胞系来 描述在细胞表面蛋白质组和分泌组中发生的全部变化的光谱 随着肿瘤从侵袭前病变发展到完全转移的疾病。在目标3中，我们将利用 组织学引导的MALDI-TOF MS组织谱识别与早期相关的生物标记物 用胰腺组织制作胰腺肿瘤动物模型。从这些结果中，我们预计 编制一套全面的检测早期胰腺癌的生物标志物。最后，在目标4中， 我们将测试在小鼠系统中识别的早期生物标记物的特异性和敏感性 使用从健康捐赠者和患者身上收集的人血清样本诊断胰腺肿瘤的能力 6例表现为慢性胰腺炎和PDAC。这些研究的目标将是开发一种 基于抗体的诊断分析，可用于筛查胰腺癌高危患者。

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.