A Novel Platform for Maternal Alcohol Consumption Screening

孕产妇酒精摄入量筛查的新平台

• 批准号: 8822061
• 负责人: Jayanth Ramadoss
• 金额: $ 23.49万
• 依托单位: TEXAS A&M AGRILIFE RESEARCH
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-05 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8822061
• 关键词: Agreement; Alcohol abuse; Alcohol consumption; Alcohols; Animal Model; Behavior Therapy; Behavioral; Biological Markers; Child; Chronic; Clinical; Collaborations; Complex; Data; Detection; Development; Diagnosis; Diagnostic; Dose; Early Diagnosis; Early identification; Fetal Alcohol Exposure; Fetal Alcohol Spectrum Disorder; Future; Genomics; Glycopeptides; Goals; Human; Individual; Knock-out; Lead; Learning Disabilities; Lectin; Link; Location; Maps; Mass Spectrum Analysis; Measures; Methodology; Methods; Modality; Modeling; Mothers; Nutritional; Oligosaccharides; Patient Self-Report; Plasma; Plasma Proteins; Polysaccharides; Post-Translational Protein Processing; Pregnancy; Pregnant Women; Prevalence; Prevention; Proteins; Proteomics; Psyche structure; Questionnaires; Rattus; Reporting; Resolution; Schools; Sialic Acids; Specificity; Strategic Planning; Technology; Testing; Time; Transferrin; Translating; United States; United States National Institutes of Health; Validation; Variant; Woman; alcohol consumption during pregnancy; alcohol effect; alcohol exposure; analytical method; base; carbohydrate-deficient transferrin; cost; disability; drinking; glycosylation; high throughput screening; high throughput technology; metabolomics; neurobehavioral; novel; pregnant; prevent; public health relevance; screening; statistics; sugar; tool; validation studies

DESCRIPTION (provided by applicant): Maternal alcohol abuse during pregnancy can result in Fetal Alcohol Spectrum Disorders (FASD), a lifelong disability characterized by a range of neuroanatomical and neurobehavioral deficits. Current estimates of FASD prevalence is about 2-5% among young school children in the United States. Therefore, early detection of alcohol use among pregnant women is critical to prevent FASD. Maternal self-reporting is largely unreliable and the utility of current biomarkers is also limited during pregnancy. Among these markers, glycosylated variants of the protein transferrin (carbohydrate deficient transferrin, CDT), is reported to be the most diagnostically specific biomarker for chronic alcohol abuse. However, the mean sensitivity is about ~65.4% with a wide range depending upon the analytical method employed. Moreover, utilizing the conventional electrophoretic, chromatographic, and immunometric methods, it has so far been difficult to characterize the composition and abundance of all glycoforms. This is critical because alcohol produces complex effects on the glycans including loss of sialic acids, partial loss of sugars, and/or complete loss of oligosaccharide chains. In a widely utilized FASD rat model, we have for the first time utilized state of the art mass spectrometry and obtained preliminary data characterizing the sequences and abundances of maternal plasma transferrin N- linked oligosaccharides preserving the location of the N-glycosite. Based on our novel data, we propose the following Specific Aim: Develop and validate a novel and sensitive high throughput platform utilizing a traditional biomarker (variants of transferrin) for maternal alcohol consumption screening in a FASD rat model. Two sub aims are proposed to characterize and validate the protein's post-translational glycosylation signature abundance profile in the maternal plasma and relating it to the dose and duration of alcohol exposure. We will utilize a pregnant rat model exposed to graded alcohol doses administered for different durations of exposure during gestation. Plasma protein (transferrin) identification, sequence mapping, glycopeptide identification with diagnostic fragmentation spectra, and High Resolution/Accurate Mass quantitation will be performed. Validation studies will include N-linked Glycosite confirmation and lectin-based probing. Based on our preliminary data, we expect the proposed platform to provide a highly sensitive protein post-translational modification signature profile that will greatly enhance the window of detection and also clearly distinguish between the alcohol doses. At the end, we plan to correlate the glycosylation signature profile with FASD neurobehavioral measures. We also believe that these novel methodologies can be in the future extended to pregnant women and form the basis for a high throughput screening tool that will help establish a standardized diagnostic criteria with a real clinical impact not only in diagnosis but also in treatment and prevention.

描述（由申请人提供）：母亲在怀孕期间酗酒可导致胎儿酒精谱系障碍 (FASD)，这是一种以一系列神经解剖学和神经行为缺陷为特征的终身残疾。目前估计美国幼儿中 FASD 的患病率约为 2-5%。因此，早期发现孕妇饮酒对于预防 FASD 至关重要。母亲的自我报告很大程度上不可靠，并且当前生物标志物在怀孕期间的效用也受到限制。在这些标记物中，据报道，转铁蛋白的糖基化变体（碳水化合物缺乏的转铁蛋白，CDT）是慢性酒精滥用诊断上最具特异性的生物标记物。然而，平均灵敏度约为 65.4%，范围很宽，具体取决于所采用的分析方法。此外，利用传统的电泳、色谱和免疫测定方法，迄今为止很难表征所有糖型的组成和丰度。这是至关重要的，因为酒精会对聚糖产生复杂的影响，包括唾液酸的损失、糖的部分损失和/或寡糖链的完全损失。在广泛使用的 FASD 大鼠模型中，我们首次利用最先进的质谱法并获得了表征保留 N-糖位点位置的母体血浆转铁蛋白 N-连接寡糖的序列和丰度的初步数据。根据我们的新数据，我们提出以下具体目标：开发并验证一种新颖且灵敏的高通量平台，利用传统生物标志物（转铁蛋白变体）在 FASD 大鼠模型中进行母体酒精消耗筛查。提出了两个子目标来表征和验证母体血浆中蛋白质的翻译后糖基化特征丰度谱，并将其与酒精暴露的剂量和持续时间联系起来。我们将利用怀孕大鼠模型，在妊娠期间暴露于不同持续时间的分级酒精剂量。将进行血浆蛋白（转铁蛋白）鉴定、序列作图、具有诊断碎片谱的糖肽鉴定以及高分辨率/精确质量定量。验证研究将包括 N-连接 Glycosite 确认和基于凝集素的探测。根据我们的初步数据，我们期望所提出的平台能够提供高度敏感的蛋白质翻译后修饰特征谱，这将大大增强检测窗口 并且还清楚地区分了酒精剂量。最后，我们计划将糖基化特征谱与 FASD 神经行为测量相关联。我们还相信，这些新方法将来可以扩展到孕妇，并构成高通量筛查工具的基础，这将有助于建立标准化的诊断标准，不仅在诊断方面而且在治疗和预防方面具有真正的临床影响。