A Lectin Glycoarray Approach for Markers of Pancreatic Cancer

描述（由申请人提供）：胰腺癌是人类恶性肿瘤中最致命的形式之一，早期发现将大大有助于提高患者的生存率。糖微阵列方法将用于在人血浆中寻找胰腺癌的早期检测标志物。我们将使用完整的N-连接的血浆糖蛋白分离凝集素亲和柱的多维液相分馏。完整糖蛋白的基于液体的分级分离最初将涉及无孔色谱法，随后是液体毛细管等电聚焦以进一步分离蛋白糖型。将每种液体级分点在硝酸纤维素包被的显微镜载玻片上以产生天然糖蛋白微阵列，然后通过各种荧光标记的凝集素询问所述天然糖蛋白微阵列以探测每个微阵列点是否存在特异性聚糖部分。将分析癌症和正常患者以及炎性病变患者的血浆样本，以寻找揭示癌症进展期间发生的特定聚糖结构变化的模式。将对来自癌症和正常患者以及相关炎性病变（慢性胰腺炎）患者的各30份样本的分析测试集进行初步分析，以识别此类潜在标志物。新的软件将用于分析这些阵列和由此产生的测试集。将鉴定与癌症相对于正常或炎症相关的糖蛋白，并使用凝集素提取/LC ESI/MS/MS和QIT-TOF（MALDI MSn）质谱法鉴定结构部分，以检查可作为癌症标志物的详细结构变化。虽然许多标记糖蛋白本身在血浆中具有相对高的丰度，但基于聚糖结构的变化，它们可能是癌症特异性的。这项研究在公共卫生领域具有重要意义，因为它可能提供一种在血浆中寻找疾病标志物的方法。这些标志物可能与疾病的阶段有关，因此可能对个性化治疗很重要，这是临床医学中的一个重要问题。

DESCRIPTION (provided by applicant): Pancreatic cancer is one of the most deadly forms of human malignancy for which early detection would greatly facilitate increased patient survival. A glyco-microarray approach will be used to search for early detection markers of pancreatic cancer in human plasma. We will use multi-dimensional liquid-phase fractionation of intact N-linked plasma glycoproteins isolated by lectin affinity columns. The liquid-based fractionation of intact glycoproteins will initially involve nonporous chromatography, followed by liquid capillary isoelectric focusing to further separate protein glycoforms. Each liquid fraction will be spotted on nitrocellulose- coated microscope slides to produce a natural glycoprotein microarray which will then be interrogated by various fluorescently-labeled lectins to probe each microarray spot for the presence of specific glycan moieties. Plasma samples from cancer and normal patients and patients with inflammatory lesions will be analyzed to search for patterns that reveal specific glycan structural changes that occur during cancer progression. Initial analysis will be performed on an analytical test set of 30 samples each from cancer and normal patients and from patients with related inflammatory lesions (chronic pancreatitis) to identify such potential markers. Novel software will be used to analyze these arrays and the resulting test set. Glycoproteins that are associated with cancer versus normal or inflammation will be identified and the structural moieties identified using lectin extraction/LC ESI/MS/MS and QIT-TOF (MALDI MSn) mass spectrometry to examine the detailed changes in structure that may serve as markers of cancer. While many of the marker glycoproteins themselves are of relatively high abundance in plasma, they may be cancer-specific based upon changes in glycan structure. This research has important implications in public health in that it may provide a methodology for searching for markers of disease in plasma. These markers may be related to the stage of the disease and, thus, may become important for personalized treatment, an important issue in clinical medicine.