Markers of Early Pancreatic Cancer

描述（由申请人提供）：本项目的总体目标是开发新的胰腺癌血清和胰液标志物，可用于诊断有疾病症状的患者，并识别早期无症状胰腺癌患者。NCI计划审查小组（PRO）将其列为研究重点-“识别和开发用于早期发现胰腺癌及其“并发症”的监测和诊断方法”。一组敏感和特异的胰腺癌标志物将大大有助于胰腺癌的早期诊断和早期发现。我们将研究三种标记策略，旨在检测血清或胰液中的胰腺肿瘤。每一个目标都集中在已经成为第2阶段研究主题的特定的、充分表征的蛋白质或DNA标记物上。一种标志物，血清MIC-1（aim #1）对CA 19 -9具有上级的总体诊断灵敏度。对于目标#2，我们将重新评估胰腺癌的一个众所周知的标志物，突变KRAS。以前的研究发现，它在胰腺中的检测缺乏必要的特异性来区分胰腺癌和胰腺炎。为了克服这种疾病特异性的缺乏，我们已经开发了一种新型的、高度敏感和特异性的测定法（“Ligands”）来检测和定量点突变，如突变型KRAS。我们的初步2期研究表明，定量突变KRAS显着提高其诊断效用。对于目标#3，我们将使用一种新的突变检测方法，当基因如p53和p16以低浓度存在于临床样品如胰液中时，该方法可以检测基因如p53和p16中的未知突变（有限稀释-PCR，或“LD-PCR”和异源双链分析（HA））。我们将3A的结果与使用p53基因芯片获得的结果进行比较。我们将追求的具体目标是：目标; 1：确定血清MIC-1的敏感性和特异性; 1A：作为早期胰腺癌的标志物。IB：作为无症状胰腺癌的预测因子，在发生这种疾病的高风险人群中，和1C;以确定不过表达MIC-1的胰腺癌的生物学和临床意义。目的2A：开发和验证5种KRAS和1种BRAF配体测定，以产生用于目的2B的组。2B：证明Ligandine检测和定量胰腺癌患者和适当对照的胰液和血清中KRAS 2和BRAF突变的诊断效用。3A;证明一种新的检测方法（有限稀释PCR或LD-PCR与异源双链分析（HA））用于检测和定量胰腺疾病或胰腺癌筛查患者的胰液中低丰度p53和p16突变的诊断效用。3B：比较目的3A（LD/PCR/HA）和p53基因芯片检测胰腺液中p53突变的能力。我们还将确定组合来自这3个目的的标记物的结果的诊断效用。

DESCRIPTION (provided by applicant): The overall goal of this project is to develop new serum and pancreatic juice markers of pancreatic cancer that can be used to diagnose patients with symptoms of their disease and to identify patients with early asymptomatic pancreatic cancer. The NCI Program review group (PRO) cited as a research priority- "to identify and develop surveillance and diagnostic methods for the early detection of pancreatic cancer and its' precursors'". A panel of sensitive and specific markers of pancreatic adenocarcinoma would greatly facilitate the early diagnosis and early detection of this disease. We will examine three marker strategies that aim to detect pancreatic neoplasia in serum or in pancreatic juice. Each aim focuses on specific, well-characterized protein or DNA markers that have already been the subject of phase 2 studies. One marker, serum MIC-1 (aim #l) has superior overall diagnostic sensitivity to CA19-9. For aim #2 we will re-evaluate a well-known marker of pancreatic cancer, mutant KRAS. Previous studies have found that its detection in the pancreas lacks the necessary specificity to differentiate pancreatic cancer from pancreatitis. To overcome this lack of disease specificity, we have developed a novel, highly sensitive and specific assay ("LigAmp") to detect and quantify point mutations such as mutant KRAS. Our preliminary phase 2 studies indicate that quantifying mutant KRAS significantly improves its diagnostic utility. For aim #3 we will use a novel mutation detection approach that can detect unknown mutations in genes such as p53 and pi 6 when they are present at low concentration in clinical samples such as pancreatic juice (limiting dilution-PCR, or "LD-PCR" and heteroduplex analysis (HA)). We will compare the results of 3A with those obtained using p53 Gene chips. The specific aims we will pursue are: Aim; 1: To determine the sensitivity and specificity of serum MIC-1; 1A: as a marker of early pancreatic cancer. IB: as a predictor of asymptomatic pancreatic cancer in a population at high risk of developing this disease, and 1C; to determine the biological and clinical significance of pancreatic cancers that do not overexpress MIC-1. Aim 2A: To develop and validate five KRAS and one BRAF LigAmp assays to produce a panel for use in aim 2B. 2B: To demonstrate the diagnostic utility of LigAmp to detect and quantify KRAS2 and BRAF mutations in pancreatic juice and in serum from patients with pancreatic cancer and appropriate controls. 3A; To demonstrate the diagnostic utility of a novel assay (Limiting dilution PCR or LD-PCR with Heteroduplex analysis (HA)) to detect and quantify low abundance p53 and pi 6 mutations in the pancreatic juice of patients with pancreatic disease or undergoing pancreatic cancer screening. 3B; To compare the results of aim 3A (LD/PCR/HA) with p53 GeneChip for their ability to detect p53 mutations in pancreatic juice. We will also determine the diagnostic utility of combining the results of the markers derived from these 3 Aims.