Proteomic and Transcriptomic Biomarkers of Circadian Timing

摘要 该项目的目标是通过开发可靠的诊断方法来优化人类的健康、性能和安全 昼夜节律的生物标记物，用于从单个生物样本中识别个体内的生物时间。 我们对昼夜节律对人类健康重要性的认识在增加；昼夜节律的扰乱 时间选择与代谢紊乱、心血管疾病、免疫失调和癌症有关。一个 最近的一项研究测试了大约17,000个基因，发现近一半的基因在至少一个人类组织中循环，而超过 其中900个循环基因编码的蛋白质是药物靶标、运输药物或与药物有关 代谢性1.在这一新兴知识的基础上，我们应该能够规律化昼夜节律，以防止 并在理想的昼夜节律时间给药，以提高疗效和/或 减少副作用。然而，我们将昼夜节律计时纳入临床决策和 由于我们目前无法快速、轻松地测量昼夜节律，治疗受到了影响。 目前评估昼夜节律的方法需要几个小时(甚至一天)的采样，而 病人处于可控状态。我们的目标是开发一种方法，可以估计个人的昼夜节律时间 在白天或晚上的任何时间采集的单一血液样本。要做到这一点，我们将使用两种最先进的方法， 一种基于血浆蛋白质组学的方法来识别一组节律蛋白质(扩展了我们的初步数据)和 一种基于全血的单核细胞方法，使用15个转录本(以验证和扩展最近的 研究)。我们将通过收集以下信息来确保选定的蛋白质生物标记物小组对不同的睡眠-觉醒模式具有健壮性 在习惯性睡眠时间、工作日睡眠时间缩短和周末睡眠时间延长条件下的样本 睡眠，以及一群健康、紧张、健康的人在大约40个小时的受控姿势、醒来和行为上 休息的成年人在高度受控的实验室条件下研究了一周。我们还将在 一系列有昼夜节律睡眠障碍的患者。我们将单独验证基于蛋白质组学的 生物标记物和基于单核细胞的转录本生物标记物，并探索结合它们是否可以改善 我们时间估计的准确性。在所有情况下，生物标志物小组对昼夜节律的估计将是 与从血浆或唾液中提取的褪黑素(目前的“黄金标准”昼夜节律阶段)相比 标记)。 这项提议的项目有可能：1)揭示生理节律影响的新的生理通路 和睡眠；2)为改善对疑似昼夜节律患者的诊断和治疗铺平道路 睡眠障碍(睡眠-觉醒延迟阶段障碍、轮班工作障碍)和其他睡眠病理(失眠、 3)通过个体化治疗时机推进个体化用药(时间医学)。

ABSTRACT The goal of the project is to optimize human health, performance, and safety by developing robust diagnostic biomarkers for circadian timing to identify, from a single biospecimen, the biological time within an individual. Our understanding of the importance of circadian timing to human health is increasing; disruption of circadian timing is associated with metabolic disorders, cardiovascular disease, immune dysregulation, and cancers. A recent study tested ~17,000 genes and found that nearly half cycled in at least one human tissue, and more than 900 of those cycling genes coded for proteins that are drug targets, transport drugs, or are involved in drug metabolism1. Building on this emerging knowledge, we should be able to regularize circadian timing to prevent disease, and to administer many short half-life drugs at their ideal circadian time to increase efficacy and/or reduce side effects. However, our ability to incorporate circadian timing into clinical decision-making and treatment is impaired by our current inability to measure circadian phase quickly and easily. Current methods for assessing circadian timing require sampling over hours (or even up to a day) while the patient is in controlled conditions. We aim to develop a method that can estimate individual circadian time with a single blood sample taken at any time of the day or night. To do this, we will use two state of the art methods, a plasma proteomics-based method to identify a panel of rhythmic proteins (extending our preliminary data) and a whole blood-derived monocyte-based method using a panel of 15 transcripts (to validate and extend a recent study). We will ensure the selected protein biomarker panel is robust to varying sleep-wake patterns by collecting samples under conditions of habitual sleep timing, under shortened “weekday” sleep and extended “weekend” sleep, and across ~40 hours of controlled posture, wake, and behaviour from a group of healthy, entrained, well- rested adults studied for a week in highly controlled laboratory conditions. We will also test both methods in a series of patients with circadian rhythm sleep disorders. We will validate separately the proteomics-based biomarker and the monocyte-based transcript biomarker, and also explore whether combining them can improve the accuracy of our timing estimates. In all cases, circadian phase estimates from the biomarker panels will be compared with those derived from plasma or saliva melatonin (the current “gold-standard” circadian phase marker). The proposed project has the potential to: 1) reveal novel physiological pathways affected by circadian timing and sleep; 2) pave the way for improved diagnosis and treatment for patients with suspected circadian rhythm disorders (delayed sleep-wake phase disorder, shift work disorder) and other sleep pathologies (insomnia, hypersomnia); 3) advance personalized medicine through individualized treatment timing (chronomedicine).