Development of the GlycoHCCTyper for the early detection of HCC

开发用于早期检测 HCC 的 GlycoHCCTyper

• 批准号: 10382624
• 负责人: Stephen Castellino
• 金额: $ 42.84万
• 依托单位: GLYCOPATH, INC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-01 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10382624
• 关键词: AFP gene; Acute Disease; Address; Algorithms; Antibodies; Biological Assay; Biological Markers; Blinded; Blood specimen; CA-125 Antigen; Ceruloplasmin; Chronic Disease; Cirrhosis; Clinical; Detection; Development; Diagnosis; Early Diagnosis; Ensure; Evaluation; FDA approved; Glycoproteins; Goals; Hand; Haptoglobins; Hemopexin; Image; Immunoglobulin G; Laboratories; Lectin; Link; Low-Molecular-Weight Kininogen; MALDI-TOF Mass Spectrometry; Malignant neoplasm of liver; Mass Spectrum Analysis; Medical; Methods; Modification; Orosomucoid; Patients; Phase; Polysaccharides; Primary Malignant Neoplasm of Liver; Primary carcinoma of the liver cells; Process; Proteins; Proteomics; Reproducibility; Sampling; Serum; Slide; Small Business Technology Transfer Research; South Carolina; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; System; Technology; Time; Tissue Sample; Transferrin; Universities; Validation; Vitamin D-Binding Protein; Work; alpha 2-Glucoproteins; alpha-Fetoproteins; apolipoprotein D; biomarker signature; cancer biomarkers; clinical development; cohort; design; glycoproteomics; glycosylation; histidine-rich glycoprotein; instrument; mass spectrometer; non-alcoholic fatty liver disease; nonalcoholic steatohepatitis; novel; prospective; protein purification; sample collection; sugar; AFP gene; Acute Disease; Address; Algorithms; Antibodies; Biological Assay; Biological Markers; Blinded; Blood specimen; CA-125 Antigen; Ceruloplasmin; Chronic Disease; Cirrhosis; Clinical; Detection; Development; Diagnosis; Early Diagnosis; Ensure; Evaluation; FDA approved; Glycoproteins; Goals; Hand; Haptoglobins; Hemopexin; Image; Immunoglobulin G; Laboratories; Lectin; Link; Low-Molecular-Weight Kininogen; MALDI-TOF Mass Spectrometry; Malignant neoplasm of liver; Mass Spectrum Analysis; Medical; Methods; Modification; Orosomucoid; Patients; Phase; Polysaccharides; Primary Malignant Neoplasm of Liver; Primary carcinoma of the liver cells; Process; Proteins; Proteomics; Reproducibility; Sampling; Serum; Slide; Small Business Technology Transfer Research; South Carolina; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; System; Technology; Time; Tissue Sample; Transferrin; Universities; Validation; Vitamin D-Binding Protein; Work; alpha 2-Glucoproteins; alpha-Fetoproteins; apolipoprotein D; biomarker signature; cancer biomarkers; clinical development; cohort; design; glycoproteomics; glycosylation; histidine-rich glycoprotein; instrument; mass spectrometer; non-alcoholic fatty liver disease; nonalcoholic steatohepatitis; novel; prospective; protein purification; sample collection; sugar

Alterations in glycosylation have long been associated with the development and progression of many types of chronic and acute diseases. Our group (Mehta) was one of the first to perform glycan analysis in serum and perform proteomics on specific glycoforms (glycoproteomics). Using such methods, we identified a number of serum glycoproteins with altered glycosylation in hepatocellular carcinoma, a primary cancer of the liver. However, because of limitations in technology, we were forced to either examine each protein one at a time or examine pools of proteins without the ability to link a particular glycan to a given protein. In some situations, we performed structural glycan analysis on purified proteins which provided the best “biomarker” information, but took days to weeks for analysis. Alternatively, we could forgo true structural information and use lectins to determine if only one specific sugar moiety was present, and perform analysis in a more rapid manner. But again, this was generally done only on one protein at a time. In all of these situations, the biomarker potential of these glycoproteins was diminished because of the technology used. To address this limitation, GlycoPath has recently developed a streamlined antibody capture slide array approach to directly profile N-linked glycans on captured serum glycoproteins. This process requires only a few microliters of sample and utilizes simple methods that require no protein purification or sugar modifications prior to analysis. This method is referred to as the GlycoTyper. In this method, N-linked glycans are released from antibody captured glycoproteins and are directly analyzed by MALDI-TOF mass spectrometry. We hypothesize that this method can be used to identify glycan biomarkers reflective of the changes that occur during the development of hepatocellular carcinoma. In this Phase I STTR application we anticipate developing a reproducible and translatable workflow using MALDI-MS of captured proteins that can accurately detect the presence of hepatocellular carcinoma.

糖基化的改变长期以来一直与 许多类型的慢性和急性疾病。我们的小组（Mehta）是第一个表演的人之一 血清中的聚糖分析并对特定糖型（糖蛋白质组学）进行蛋白质组学。使用 这种方法，我们鉴定了许多血清糖蛋白，并在 肝细胞癌，肝脏的原发性癌。 但是，由于技术的局限性，我们被迫检查每种蛋白质 一次一个或检查蛋白质池，没有能力将特定的聚糖连接到给定的 蛋白质。在某些情况下，我们对纯化蛋白进行了结构性聚糖分析，该蛋白质的结构性分析 提供了最佳的“生物标志物”信息，但花了数天到几周才能进行分析。另外，我们 可能会忘记真正的结构信息，并使用讲座来确定是否只有一种特定的糖 存在部分，并以更快的方式进行分析。但同样，这通常是 一次仅在一种蛋白质上完成。在所有这些情况下，这些情况的生物标志物潜力 由于使用的技术，糖蛋白降低了。 为了解决这一限制，糖原最近开发了一种简化的抗体捕获幻灯片 阵列方法在捕获的血清糖蛋白上直接介绍N连接的聚糖。这个过程 仅需几个微量的样品，并且使用不需要蛋白质的简单方法 分析前进行纯化或糖修饰。该方法称为糖蛋白酶。 在这种方法中，N连接的聚糖是从捕获的糖蛋白中释放出来的，并且是 通过MALDI-TOF质谱法直接分析。我们假设这种方法可以是 用于识别反映在开发过程中发生的变化的聚糖生物标志物 肝细胞癌。在此阶段，我预计会开发一个 使用捕获的蛋白质的MALDI-MS可重现和可翻译的工作流程，可以准确地 检测肝细胞癌的存在。