Serum Glyco-Markers of Early Hepatocellular Carcinoma Using a Mass Spec Approach

使用质谱方法检测早期肝细胞癌的血清糖标记物

• 批准号: 10285013
• 负责人: David M. Lubman
• 金额: $ 38.43万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10285013
• 关键词: Abdomen; Address; Archives; Biological Assay; Biological Markers; Blinded; Cancer Etiology; Cessation of life; Characteristics; Cirrhosis; Clinical; Clinical Data; Clinical Treatment; Complex; Computer software; Coupled; Data; Databases; Detection; Disease; Early Detection Research Network; Early Diagnosis; Enzyme-Linked Immunosorbent Assay; Ethnic Origin; Etiology; Glycopeptides; Glycoproteins; Goals; Guidelines; Haptoglobins; Incidence; Laboratories; Lectin; Liver diseases; Malignant Neoplasms; Mass Spectrum Analysis; Measurement; Methods; Minor; Monitor; Patients; Peptides; Performance; Phase; Polysaccharides; Primary carcinoma of the liver cells; Professional Organizations; Proteins; Publishing; Race; Reaction; Sampling; Screening for cancer; Serum; Site; Specificity; Structure; Survival Rate; Techniques; Testing; Ultrasonography; Validation; Work; alpha-Fetoproteins; base; bioinformatics tool; biomarker performance; biomarker validation; carbohydrate structure; curative treatments; detection sensitivity; early detection biomarkers; glycoproteomics; glycosylation; high risk; improved; innovation; multiplex assay; novel; screening; sex; sialylation; success; surveillance strategy; tandem mass spectrometry

Abstract: Hepatocellular carcinoma (HCC) is the fourth most common cause of cancer-related death worldwide and is rising in incidence in the US. Over 80% of patients in the US with HCC have underlying cirrhosis, and professional societies guidelines recommend surveillance in all patients with cirrhosis to facilitate early detection. Unfortunately, only 20-30% of patients are detected at an early stage when potentially curative treatments are possible. Our proposal, based on our strong preliminary data evaluating glycoproteomic profiles, addresses the clear unmet need for accurate early HCC detection biomarkers. Unique changes in glycosylation in proteins, which involve structural changes in glycan groups, have been shown to be important serum biomarkers for early HCC detection. Importantly, even subtle changes involving minor structures can be very specific in differentiating cirrhosis versus early stage HCC. In this study, we will use a mass spectrometry approach, which has proven to be a highly sensitive and accurate way to detect and quantitatively monitor glycan structural changes. Our published mass analysis discovery work, tandem mass spectrometry measurements, and novel bioinformatic tools for glycan and glycopeptide analysis demonstrate these minor changes in structure can act as promising early HCC detection biomarkers. In aim 1 these methods will be demonstrated for glycopeptide screening from serum using novel mass spec assays using parallel reaction monitoring (PRM) analysis based on a stepped HCD fragmentation method which can ultimately be used to distinguish early stage HCC from cirrhosis. The PRM method is a targeted assay which can distinguish small changes in glycan structure and can be multiplexed to monitor large numbers of markers simultaneously. In addition, using separations coupled to the PRM strategy, we will be able to distinguish different isomeric forms of fucosylation and sialylation in glycan structures which may contain important disease related markers. In aim 2 the methods developed herein will be optimized and then applied to a large confirmation set of 300 early stage HCC and cirrhosis samples for 20 target glycopeptides from Haptoglobin and to several other potential glycopeptide markers discovered and evaluated in our prior work. In aim 3 we will then validate the biomarker performance in a blinded phase II biomarker study using a large set of 800-1000 early stage HCC and cirrhosis patients, diverse with regard to sex, race/ethnicity, and liver disease etiology. Through these aims, we will identify a panel of glycopeptides that can serve as highly accurate biomarkers for early HCC detection.

摘要：肝细胞癌(Hcc)是全球第四大癌症相关死亡原因。 在美国的发病率正在上升。在美国，超过80%的肝细胞癌患者有潜在的肝硬变， 专业协会指南建议对所有肝硬变患者进行监测，以便于早期发现。 不幸的是，只有20%-30%的患者在早期阶段被发现，当时潜在的治愈方法是 有可能。我们的建议，基于我们评估糖蛋白质组谱的强大初步数据，解决了 明确尚未满足的对准确的肝细胞癌早期检测生物标志物的需求。蛋白质中糖基化的独特变化， 涉及多糖基团的结构变化，已被证明是早期重要的血清生物标志物 肝细胞癌检测。重要的是，即使是涉及微小结构的细微变化也可以在区分方面非常具体 肝硬变与早期肝细胞癌。在这项研究中，我们将使用质谱学方法，这已被证明 是检测和定量监测多糖结构变化的一种高度灵敏和准确的方法。我们的 已发表的质量分析发现工作、串联质谱学测量和新的生物信息学 糖链和糖肽分析工具表明，这些微小的结构变化可能是有希望的 肝细胞癌早期检测生物标志物。在目标1中，这些方法将用于糖肽的筛选 采用基于STEP的平行反应监测(PRM)分析的新型质谱仪分析血清 HCD碎片法，最终可用于区分早期肝细胞癌和肝硬变。这个 PRM方法是一种靶向分析方法，可以区分糖链结构的微小变化，并且可以进行多重分析 同时监测大量的标志物。此外，使用与PRM策略相结合的分离， 我们将能够区分多糖结构中岩藻糖基化和唾液酸化的不同异构体 可能包含重要的疾病相关标志物。在目标2中，这里开发的方法将被优化 然后应用于300个早期肝细胞癌和肝硬变样本的20个目标的大确认集 结合珠蛋白糖肽和其他几种潜在糖肽标记物的发现和评价 在我们之前的工作中。在目标3中，我们将在II期盲法研究中验证生物标记物的性能 使用800-1000名早期肝细胞癌和肝硬变患者的大集合，这些患者在性别、种族/民族、 和肝病病因学。通过这些目标，我们将确定一组可以高度 用于肝细胞癌早期检测的准确生物标志物。