Supplemental for Detection of Glycopeptides of MCI in Patient Serum

摘要：阿尔茨海默病(AD)是一种神经退行性疾病，占痴呆的大部分 病例，这影响到全球约3000万患者。轻度认知障碍(MCI)已被认识到 作为进展期AD前临床损害的中间状态。由于阿尔茨海默病引发的大脑变化 在出现初始症状之前，如MCI，需要早期诊断生物标志物。 研究。然而，由于缺乏确定性，AD的早期诊断是一个巨大的挑战 生物标记物，与其他类似神经退行性疾病重叠的生物标记物，以及发现 检测这些生物标志物的微创方法。已经证明，人类社会的独特变化 涉及糖基团结构变化的蛋白质的糖基化可能是重要的血清生物标志物 用于早期发现各种疾病。我们已经从患者血清中识别出了这种潜在的糖肽， 这可能有助于识别早期MCI的发展。因此，在拟议的工作中，我们将采用有针对性的 基于细节的质谱法筛选患者血清中MCI与正常对照的标志物 多聚糖的结构及其位置特异性。这将基于我们使用多路并行反应的目标 监测(PRM-MS)定量检测目标糖肽，我们可以监测多达50个目标 标记同时出现。然后，我们将使用潜在的PRM分析来演示初步的确认研究 糖肽标记物可以区分正常和MCI患者并决定其表现 根据每个标记物的敏感度/特异度。这项工作将导致糖肽生物标记物的早期阶段 使用多路PRM-MS方法的MCI，这些标记物将用于进一步的临床 验证。此外，我们还将用其他神经退行性疾病的样本来测试这种方法。

Abstract: Alzheimer’s disease (AD) is a neurodegenerative disorder that accounts for the majority of dementia cases, which affects around 30 million patients worldwide. Mild cognitive impairment (MCI) has been recognized as an intermediate state of clinical impairment before advanced AD. Due to changes in the brain triggered by AD before the presentation of initial symptoms, such as for MCI, there is a need for early-stage diagnosis biomarker research. However, early-stage diagnosis of AD represents a significant challenge due to a lack of definitive biomarkers, an overlap of biomarkers with other similar neurodegenerative diseases, and the ability to find minimally invasive methods for detection of these biomarkers. It has been shown that unique changes in glycosylation in proteins that involve structural changes in glycan groups may be important as serum biomarkers for early detection of various diseases. We have identified such potential glycopeptides from patient serum, which may serve to identify early-stage MCI development. In the proposed work we will thus employ a targeted mass spectrometry approach to screen patient serum for markers of MCI versus normal based on the detailed structure of glycans and their site specificity. This will be based on our aim to use a multiplexed Parallel Reaction Monitoring (PRM-MS) assay to quantitatively detect targeted glycopeptides where we can monitor up to 50 target markers simultaneously. We will then demonstrate an initial confirmation study using the PRM assay of potential glycopeptide markers that can discriminate among normal versus MCI patients and determine the performance of each marker based on its sensitivity/specificity. This work will result in glycopeptide biomarkers of early-stage MCI using a multiplexed PRM-MS method where these markers will then be available for further clinical validation. In addition, we will also test this method against samples from other neurodegenerative diseases.