Serum glycoprotein markers of cancer using an ion mobility/mass spec approach

使用离子淌度/质谱方法测定癌症的血清糖蛋白标记物

• 批准号: 8019264
• 负责人: David M. Lubman
• 金额: $ 31.29万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8019264
• 关键词: Address; Adult; Affinity; Antibodies; Antibody Affinity; Archives; Benign; Binding; Biological Assay; Biological Markers; Blinded; Cancer Etiology; Cessation of life; Clinical; Collection; Colorectal Cancer; Cystic Neoplasm; Data; Decision Making; Detection; Diagnostic; Disease; Early Diagnosis; Epitopes; Excision; Fractionation; Future; Gender; Glycoproteins; Incidence; Inflammatory; Label; Lead; Lectin; Link; Liquid substance; Malignant Neoplasms; Malignant neoplasm of esophagus; Malignant neoplasm of ovary; Malignant neoplasm of pancreas; Masks; Mass Spectrum Analysis; Methodology; Methods; Minority; Monitor; Mucinous; Mucinous Neoplasm; Non-Insulin-Dependent Diabetes Mellitus; Pain; Pancreas; Pancreatic Adenocarcinoma; Pancreatic carcinoma; Papillary; Patients; Pattern; Phase; Polysaccharides; Primary carcinoma of the liver cells; Production; Protein Glycosylation; Proteins; Resectable; Sampling; Screening for cancer; Sensitivity and Specificity; Serum; Serum Proteins; Specificity; Staging; Structure; Survival Rate; Techniques; Testing; United States; Unresectable; Validation; Work; base; carbohydrate structure; chronic pancreatitis; glycosylation; high throughput analysis; ion mobility; mortality; outcome forecast; success; sugar; tandem mass spectrometry; tool; validation studies

DESCRIPTION (provided by applicant): Pancreatic adenocarcinoma is the 4th leading cause of cancer-related death in the United States (1) and is associated with a 4% five-year survival rate. Most patients have unresectable disease at presentation although a small minority of patients (10-20%) is found to have early resectable pancreatic cancer. A method for early-stage detection of pancreatic cancer would provide a prospect of long-term survival and even curability. A highly specific noninvasive test that can differentiate early pancreatic adenocarcinoma from unresectable disease, as well as chronic pancreatitis and other non-pancreatic cancers and inflammatory conditions may provide a valuable clinical tool. Many types of malignancies have been shown to possess unique glycan moieties on specific glycoproteins in comparison to the same proteins found in normal states or other diseases that can be detected in the serum. In the proposed work, we will use a mass spec based assay to enhance the analytical specificity for detecting changes in glycan structures between disease states. We will use a platform involving an antibody capture of selected proteins from serum followed by deglycosylation and collection of the glycans from these proteins. The glycans will then be analyzed by an ion mobility/tandem mass spectrometry method which is sensitive to even subtle changes in glycan structure. The ion mobility method can provide a rapid separation of the glycans including glycoforms and the use of tandem mass spec detection can readily observe changes in the glycan structure at the sugar level as a function of disease. The method can be multiplexed to analyze several glycoprotein changes simultaneously where it can be used for high-throughput analysis of a large number of samples using the combined separation capabilities of the ion mobility method and mass spec techniques. The proposed work will involve studying fifteen antibody/protein combinations which were selected based on our preliminary glycoarray studies. We hypothesize that unique glycosylation patterns can be observed in proteins from serum samples of patients with benign pancreatic mucinous cystic neoplasms (MCNs) and intraductal papillary mucinous neoplasms (IPMNs), patients with long-term type II diabetes, patients with chronic pancreatitis, and patients with pancreatic cancer. To address this hypothesis, we will use 50 serum samples from each of the five disease groups to calculate sensitivity and specificity, to identify markers that should be advanced to a blinded test set and to define cut- points for decision making. We will also screen for comparison 50 samples each of other cancers that are archived including esophageal, colorectal, hepatocellular, and ovarian cancers. We hypothesize that the glycosylation patterns of pancreatic cancer are unique and do not overlap with other cancers. PUBLIC HEALTH RELEVANCE: If this work is successful then it will lead to potential markers that can be used for future pre-validation studies for the early detection of pancreatic cancer based upon an antibody/ion mobility/tandem mass spectrometry assay that will reveal unique changes that can be detected in patient serum.

描述（由申请人提供）：胰腺癌是美国癌症相关死亡的第四大原因 (1)，五年生存率为 4%。尽管少数患者（10-20%）被发现患有早期可切除的胰腺癌，但大多数患者就诊时患有不可切除的疾病。一种早期检测胰腺癌的方法将为长期生存甚至治愈提供前景。一种高度特异性的非侵入性测试可以区分早期胰腺腺癌和不可切除的疾病，以及慢性胰腺炎和其他非胰腺癌和炎症性疾病，可能提供有价值的临床工具。与正常状态或可在血清中检测到的其他疾病中发现的相同蛋白质相比，许多类型的恶性肿瘤已被证明在特定糖蛋白上拥有独特的聚糖部分。在拟议的工作中，我们将使用基于质谱的测定来增强检测疾病状态之间聚糖结构变化的分析特异性。我们将使用一个平台，该平台涉及从血清中捕获选定蛋白质的抗体，然后进行去糖基化并从这些蛋白质中收集聚糖。然后通过离子淌度/串联质谱法对聚糖进行分析，该方法对聚糖结构的细微变化也很敏感。离子淌度方法可以快速分离包括糖型在内的聚糖，并且使用串联质谱检测可以轻松观察糖水平上聚糖结构随疾病的变化。该方法可以多重使用，同时分析多种糖蛋白变化，利用离子淌度法和质谱技术的组合分离能力，可用于大量样品的高通量分析。拟议的工作将涉及研究十五种抗体/蛋白质组合，这些组合是根据我们初步的糖阵列研究选择的。我们假设，在良性胰腺粘液性囊性肿瘤（MCN）和导管内乳头状粘液性肿瘤（IPMN）患者、长期II型糖尿病患者、慢性胰腺炎患者和患者的血清样本中可以观察到独特的糖基化模式。患有胰腺癌。为了解决这一假设，我们将使用来自五个疾病组的每一个的 50 个血清样本来计算敏感性和特异性，以确定应进入盲法测试集的标记物，并定义决策的切入点。我们还将筛选比较 50 个已存档的其他癌症样本，包括食管癌、结直肠癌、肝细胞癌和卵巢癌。我们假设胰腺癌的糖基化模式是独特的，并且与其他癌症不重叠。 公共健康相关性：如果这项工作成功，那么它将产生潜在的标记物，可用于未来的预验证研究，以基于抗体/离子淌度/串联质谱分析来早期检测胰腺癌，从而揭示独特的变化可以在患者血清中检测到。