Developing high-throughput IMS-MS and IMS-IMS-MS techniques for glycomics analysi

描述（由申请人提供）：在过去的二十年中，美国食管腺癌（EAC）发病率的增加代表了一个重大的健康挑战。考虑到接受EAC治疗的患者的高死亡率，这一点尤其明显。目前科学研究的目标是确定与EAC相关的分子标记。考虑到碳水化合物在细胞过程中的多元作用，糖组学是一个受到特别关注的领域。比较糖组学分析的一个限制因素是假定存在于生物样品中的无数聚糖结构，这对分析化学家在组分分辨率和鉴定方面提出了挑战。这对于基于质谱（MS）的分析平台来说尤其成问题，因为单靠质谱无法实现同分异构体的分辨率。在这里，我们提出使用离子迁移率光谱（IMS）技术结合质谱快速表征血浆聚糖消化。具体而言，多维IMS （IMS-IMS）方法将被开发，以提供等离子体样品的最高效率表征。IMS-IMS与质谱的结合可以快速分辨多糖异构体。这种使能技术允许对生物标志物验证所需的数百个血浆样品进行高通量比较。作为研究的一部分，新开发的技术将应用于使用1000个血浆样本的群体研究来验证候选聚糖的生物标志物。这里提出的工作可能对疾病诊断以及追踪与疾病进展相关的生理变化（或治疗导致的回归）的能力具有巨大的意义。此外，信息丰富的数据集也可能为疾病的分子因果机制提供线索。

DESCRIPTION (provided by applicant): The increased incidence of Esophageal Adenocarcinoma (EAC) in the United States over the past two decades represents a significant health challenge. This is especially evident considering the high mortality rate of patients who have undergone treatment for EAC. A current goal of scientific research is to identify molecular markers associated with EAC. Considering the multivariate roles of carbohydrates in cellular processes, one field receiving particular attention is glycomics. A limiting factor for comparative glycomics profiling is the myriad glycan structures postulated to exist in biological samples which present challenges for analytical chemists in the form of component resolution and identification. This is especially problematic for mass spectrometry (MS)-based analytical platforms because isomer resolution cannot be achieved with MS alone. Here we propose the use of ion mobility spectrometry (IMS) techniques combined with MS for the rapid characterization of plasma glycan digests. Specifically, multidimensional IMS (IMS-IMS) methods will be developed to provide the highest efficiency characterization of plasma samples. The combination of IMS-IMS with MS allows for rapid resolution of glycan isomers. This enabling technology allows for high-throughput comparison of hundreds of plasma samples necessary for biomarker validation. As part of the research proposed here, the newly developed technology will be applied to biomarker validation of glycan candidates using a population study of 1000 plasma samples. The work proposed here could have tremendous implications for disease diagnostics as well as the ability to track physiological changes associated with disease progression (or regression resulting from therapy). In addition it is possible that the information-rich datasets will also provide clues into molecular causal mechanisms of disease. PUBLIC HEALTH RELEVANCE: The proposed research will develop ion mobility spectrometry (IMS) techniques coupled with mass spectrometry (MS) for comparative glycomics analyses of human plasma. The new technology provides the greatest ability to resolve glycan isomers and thus correlate changes in specific structures with phenotypic differences. The approach will be used to discover and validate new glycan biomarkers for esophageal adenocarcinoma (EAC).