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 并且在美国发病率正在上升。在美国，超过 80% 的 HCC 患者有潜在的肝硬化，并且 专业协会指南建议对所有肝硬化患者进行监测，以促进早期发现。 不幸的是，只有 20-30% 的患者能够在早期阶段发现潜在的治愈性治疗。 可能的。我们的建议基于评估糖蛋白组谱的强大初步数据，解决了 对准确的早期 HCC 检测生物标志物的需求显然尚未得到满足。蛋白质糖基化的独特变化， 涉及聚糖基团的结构变化，已被证明是早期诊断的重要血清生物标志物。 肝癌检测。重要的是，即使是涉及较小结构的细微变化也可以非常具体地区分 肝硬化与早期肝癌。在本研究中，我们将使用质谱方法，该方法已被证明可以 是一种高度灵敏且准确的检测和定量监测聚糖结构变化的方法。我们的 发表的质量分析发现工作、串联质谱测量和新颖的生物信息学 聚糖和糖肽分析工具表明这些结构上的微小变化可能具有前景 早期 HCC 检测生物标志物。在目标 1 中，这些方法将被证明可用于糖肽筛选 使用基于步进的平行反应监测 (PRM) 分析的新颖质谱分析来检测血清 HCD 碎片方法最终可用于区分早期 HCC 和肝硬化。这 PRM 方法是一种靶向检测方法，可以区分聚糖结构的微小变化，并且可以进行多重检测 同时监测大量标记。此外，使用与 PRM 策略相结合的分离， 我们将能够区分聚糖结构中岩藻糖基化和唾液酸化的不同异构形式 其中可能包含重要的疾病相关标记。在目标 2 中，本文开发的方法将得到优化 然后应用于 20 个目标的 300 个早期 HCC 和肝硬化样本的大型确认集 发现并评估了触珠蛋白的糖肽和其他几种潜在的糖肽标记物 在我们之前的工作中。在目标 3 中，我们将在盲法 II 期生物标志物研究中验证生物标志物的性能 使用大量 800-1000 名早期 HCC 和肝硬化患者，这些患者在性别、种族/民族方面各不相同， 和肝脏疾病的病因学。通过这些目标，我们将确定一组可以高度发挥作用的糖肽。 用于早期 HCC 检测的准确生物标志物。