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: 10657544
• 负责人: David M. Lubman
• 金额: $ 35.82万
• 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/10657544
• 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; Isomerism; 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; Recommendation; Sampling; Screening for cancer; Serum; Site; Specificity; Structure; Survival Rate; Techniques; Testing; Validation; Work; alpha-Fetoproteins; 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; ultrasound

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）是全球与癌症相关死亡的第四个最常见原因 并正在美国发病率上升。在美国，超过80％的HCC患者患有肝硬化，并且 专业社会指南建议对所有肝硬化患者进行监视，以促进早期发现。 不幸的是，在潜在的治疗性治疗是在早期阶段仅检测到20-30％的患者 可能的。我们的建议基于我们的强大初步数据评估糖蛋白质组概况，以解决 明确的未满足的需求是准确的早期HCC检测生物标志物。蛋白质中糖基化的独特变化， 涉及聚糖组的结构变化，已被证明是早期的重要血清生物标志物 HCC检测。重要的是，即使涉及次要结构的微妙变化也可以在区分方面非常具体 肝硬化与早期HCC。在这项研究中，我们将使用一种质谱法，已证明 是一种高度敏感和准确的方法，可以检测和定量监测聚糖结构变化。我们的 已发表的质量分析发现工作，串联质谱测量和新型生物学 聚糖和糖肽分析的工具表明，结构上的这些微小变化可以是有希望的 早期HCC检测生物标志物。在AIM 1中，将证明这些方法用于糖肽筛查 使用平行反应监测（PRM）分析的新型质量规格测定的血清基于阶梯 HCD破碎方法最终可用于区分早期HCC和肝硬化。这 PRM方法是一种有针对性的测定法，可以区分聚糖结构的小变化，并且可以多重 同时监视大量标记。另外，使用与PRM策略耦合的分离， 我们将能够区分聚糖结构中不同的异构体形式 可能包含重要的疾病相关标记。在AIM 2中，本文开发的方法将被优化 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 发现和评估的其他几种潜在的糖肽标记物中的糖肽和其他几种潜在的糖肽标记物 在我们的先前工作中。在AIM 3中，我们将在II期生物标志物研究中验证生物标志物的性能 使用大量的800-1000阶段的HCC和肝硬化患者，在性别，种族/种族方面有所不同 和肝病病因。通过这些目标，我们将确定一组糖肽，这些糖肽可以很高 适用于早期HCC检测的精确生物标志物。

项目成果

GlycoSLASH: Concurrent Glycopeptide Identification from Multiple Related LC-MS/MS Data Sets by Using Spectral Clustering and Library Searching.

• DOI: 10.1021/acs.jproteome.3c00066
• 发表时间: 2023-05-05
• 期刊: JOURNAL OF PROTEOME RESEARCH
• 影响因子: 4.4
• 作者: Li, Sujun;Zhu, Jianhui;Lubman, David M.;Zhou, He;Tang, Haixu
• 通讯作者: Tang, Haixu

Large Scale Screening and Quantitative Analysis of Site-Specific N-Glycopeptides from Human Serum in Early Alzheimer's Disease Using LC-HCD-PRM-MS.

• DOI: pii: 587
• 发表时间: 2022
• 期刊: Journal of proteomics & bioinformatics
• 作者: Pan L;Lin Y;Zhu J;Zhang J;Tan Z;Lubman DM
• 通讯作者: Lubman DM

The analysis of alpha-1-antitrypsin glycosylation with direct LC-MS/MS.

使用直接 LC-MS/MS 分析 Alpha-1-抗胰蛋白酶糖基化

• DOI: 10.1002/elps.201700426
• 发表时间: 2018-09
• 期刊: Electrophoresis
• 影响因子: 2.9
• 作者: Yin H;An M;So PK;Wong MY;Lubman DM;Yao Z
• 通讯作者: Yao Z

David M. Lubman-The University of Michigan-A retrospective in research.

• DOI: 10.1002/mas.21718
• 发表时间: 2023-03
• 期刊: Mass spectrometry reviews
• 影响因子: 6.6
• 作者: Lubman DM

Rapid separation of blood plasma exosomes from low-density lipoproteins via a hydrophobic interaction chromatography method on a polyester capillary-channeled polymer fiber phase.

• DOI: 10.1016/j.aca.2021.338578
• 发表时间: 2021-07-04
• 期刊: Analytica chimica acta
• 影响因子: 6.2
• 作者: Huang S;Ji X;Jackson KK;Lubman DM;Ard MB;Bruce TF;Marcus RK
• 通讯作者: Marcus RK