Screening of Glycan Markers in Serum for Early Detection of HCC in Different Etiologies of Disease

摘要：肝细胞癌是癌症相关死亡的第三大常见原因。 在全球范围内，并且在美国的发病率正在上升。在美国，90%的肝癌患者有潜在的 因此，指南建议建议对所有肝硬变患者进行监测，以促进早期 侦测。不幸的是，只有20%-30%的患者被及早发现，因此有资格 有可能治愈的疗法。肝癌对可靠的生物标志物的需求尚未得到满足，以促进 坚持筛查，及早发现。在拟议的工作中，我们将开发早期检测 基于糖蛋白图谱的肝细胞癌治疗策略。蛋白质糖基化的独特变化，这是 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 癌症检测。重要的是，细微的变化可能只涉及微小的结构，但它们可能非常重要 在区分肝硬变与早期和晚期肝细胞癌方面具有特异性。此外，这些变化可能是 具体到肝病的病因学。这些糖链结构的变化将被检测和监测 使用已被证明是一种准确的表征方法的质谱学方法进行定量 即使是结构上的微小变化，作为基于我们先前质量的生物标记物也可能是重要的 已为多糖开发的分析、串联质谱学测量和数据库 和糖肽分析。这一点将用于从血清中筛选糖链和糖肽。 使用与质谱学相结合的新的提取和分离方法，最终可用于 鉴别早期肝细胞癌与肝硬变。拟议中的工作将提供分离和质量分析 方法实现了多糖和糖肽的异构体分离，允许明确指定 与疾病状态有关的蛋白质糖基化。我们将能够区分不同的异构体形式 岩藻糖化和唾液酸化，可能包含重要的疾病相关标志物。新的软件将是 开发并用于指认这些糖链结构。这些标记将被发现用于特定的病因 丙型肝炎病毒相关的、酒精相关的和非酒精性脂肪肝相关的肝细胞癌的病因。这将是一项多点研究，包括 肿瘤生物标记物实验室所需的所有组件，包括样品和样品制备、分离 以及质谱学分析、生物信息学评价和统计分析。最终，我们将发展 从患者血清中发现糖链/糖肽标记物的方法及其潜在的鉴定 标记物和新检测方法的发展，以提供对这些标记物的有限确认。

Abstract: Hepatocellular carcinoma (HCC) is the third most common cause of cancer-related death worldwide and is rising in incidence in the US. 90% of patients in the US with liver cancer have underlying cirrhosis, thus guideline recommendations recommend surveillance in all patients with cirrhosis to facilitate early detection. Unfortunately, only 20-30% of patients are detected with early detection and are thus eligible for potentially curative treatments. There is an unmet need for reliable biomarkers for HCC to facilitate adherence to screening and for early detection. In the proposed work we will develop early detection strategies for HCC based on glycoproteomic profiles. Unique changes in glycosylation in proteins, which involve structural changes in glycan groups, have been shown to be important serum biomarkers for early cancer detection. Importantly, the subtle changes may only involve minor structures but they can be very specific in differentiating cirrhosis versus early versus late stage HCC. In addition, these changes may be specific to the etiology of liver disease. These glycan structural changes will be detected and monitored quantitatively using a mass spectrometry approach which has proven to be an accurate way to characterize even minor changes in structure which may be significant as biomarkers based on our previous mass analysis, tandem mass spectrometry measurements and databases which have been developed for glycan and glycopeptide analysis. This will be demonstrated for both glycan and glycopeptide screening from serum using novel extraction and separation methods coupled to mass spectrometry which can ultimately be used to distinguish early stage HCC from cirrhosis. The proposed work will deliver separations and mass spec methods enabling isomeric separation of glycans and glycopeptides, permitting unequivocal assignment of protein glycosylation related to disease state. We will be able to distinguish different isomeric forms of fucosylation and sialylation which may contain important disease related markers. Novel software will be developed and used to assign these glycan structures. The markers will be discovered for specific etiologies of HCV-related, alcohol-related and NAFLD-related etiologies of HCC. This will be a multisite study to include all components required for a tumor biomarker lab including samples and sample preparation, separations and mass spec analysis, bioinformatics evaluation and statistical analysis. Ultimately, we will develop methods for discovery of glycan/glycopeptide markers from patient serum, the identification of potential markers and the development of new assays to provide a limited confirmation of these markers.