Biomarker for Sleep Loss: A Proteomic Determination

描述（由申请人提供）：本申请涉及广泛的挑战领域（03）：生物标志物发现和验证以及特定的挑战主题，03-HL-101：鉴别和验证血液、血管、心脏和呼吸道功能障碍的诊断和治疗反应的临床相关、可量化的生物标志物。睡眠不足在美国人中很常见。一些群体，如卫生保健工作者（医生，护士），由于他们的工作时间表，睡眠不足的风险特别高。睡眠不足对认知能力有重大影响，增加了车祸和医疗差错的风险。它还导致葡萄糖处理异常（胰岛素抵抗），肥胖率增加，并且是心血管疾病的危险因素。然而，个体之间对睡眠不足的反应存在重大差异，最近的数据表明这种差异在很大程度上是遗传的。虽然这些后果和个体差异是已知的，但目前还没有经过验证的生物标志物来评估睡眠不足。该提案计划利用一种新的蛋白质组学方法来识别睡眠不足的生物标志物。研究将在健康志愿者中进行，这些志愿者在正常的一天中每4小时采集一次血液样本，以及在渐进性睡眠剥夺期间和恢复睡眠期间。这些人类研究已经在10名对睡眠剥夺的影响相对抵抗的受试者（低反应者）和10名表现出明显行为反应的受试者（高反应者）中进行。 公共卫生相关性：最先进的蛋白质组学方法将应用于这些已经收集的血浆样本，以解决两个基本问题：（1）在睡眠剥夺的情况下，控制昼夜节律的影响，血液中的蛋白质是否会发生变化，这些变化可以作为睡眠丧失的生物标志物;以及（2）候选生物标志物随睡眠丧失的变化幅度在对睡眠丧失具有低响应的受试者与对睡眠丧失具有高响应的受试者之间是否不同？为了解决这些问题，我们将使用一种深入的定量蛋白质组学方法，结合同位素标记，多维蛋白质鉴定技术（MudPIT）协议和质谱法，用于高置信度鉴定睡眠剥夺受试者血浆中的蛋白质变化。拟议的研究旨在确定睡眠不足的生物标志物。我们假设有分子特征反映了困倦的状态。我们将使用一种新的蛋白质组学方法来识别对睡眠不足做出反应的蛋白质。

DESCRIPTION (provided by applicant): This application addresses broad Challenge Area (03): Biomarker Discovery and Validation and specific Challenge Topic, 03-HL-101: Identify and validate clinically relevant, quantifiable biomarkers of diagnostic and therapeutic responses for blood, vascular, cardiac, and respiratory tract dysfunction. Sleep loss is common in the American population. Some groups, such as health care workers (physicians, nurses), are particularly at risk for sleep loss because of their work schedules. Sleep loss has a major impact on cognitive performance with an increased risk of vehicular crashes, and medical errors. It also results in abnormalities in glucose handling (insulin resistance), increased obesity rates, and is a risk factor for cardiovascular disease. There are, however, major differences in the response to sleep loss between individuals and recent data indicate that this difference is in large part genetic. While these consequences and individual differences are known, there is currently no validated biomarker to assess sleep loss. This proposal plans to utilize a novel proteomic approach to identify biomarkers for sleep loss. Studies will be done in healthy volunteers, who had blood samples taken every 4 hours during a normal day, as well as during progressive sleep deprivation, and during recovery sleep. These human studies have already been conducted in 10 subjects who are relatively resistant to the effects of sleep deprivation (low responders) and 10 who show a marked behavioral response (high responders). PUBLIC HEALTH RELEVANCE: A state of the art proteomic approach will be applied to these already collected plasma samples to address two fundamental questions: (1) are there changes in proteins in blood with sleep deprivation, controlling for circadian influences, that can act as biomarkers for sleep loss; and (2) is the magnitude of change with sleep loss in candidate biomarkers different between subjects with a low response to sleep loss to those with a high response? To address these questions we will use an in-depth quantitative proteomic approach, combining isotopic labeling, a multidimensional protein identification technology (MudPIT) protocol and mass spectrometry, for high confidence identification of protein changes in the plasma of sleep deprived subjects. The proposed studies are designed to identify a biomarker for sleep loss. We hypothesize that there are molecular signatures that reflect the state of sleepiness. We will use a novel proteomic approach to identify proteins that change in response to sleep loss.