The metabolomic study of healthy ageing applying dried blood and urine spot collection

背景和科学问题--英国人口寿命的增长速度超过了健康寿命，导致晚年健康状况不佳的时间更长，并给NHS带来了相关的经济压力。有必要改善英国人口的健康老龄化。久坐的行为与运动的作用被证明对健康的衰老-新陈代谢轴有很大的影响；运动是一种积极的生活方式干预，对所有人都是可用的。代谢变化可以通过对数千种代谢物的非靶向性代谢组学分析来研究。在这些研究中，生物流体样本在一个专门的中心收集，这需要受试者出差，需要专家人员和仪器来收集和处理样本；这一过程昂贵且效率低下。然而，远离专门中心的样本收集将降低样本收集成本，并使纵向研究得以廉价进行，并将对受试者的不便降至最低。在远离专业中心的地方采集干燥的血液或尿点(DBS/DUS)，然后进行代谢组学分析，提供了一个简单而可靠的解决方案。这些设备目前被应用于靶向代谢物分析，我们希望开发这项技术用于健康年龄的非靶向代谢组学研究。SCIENTIFIFIC目标-我们建议开发和应用非靶向代谢组学的干血和尿点采集应用于健康年龄的研究。目的1-发展干血和尿点的非靶向代谢组学分析方法。与检测到的代谢物类别的灵敏度和范围相关的小样本量(通常为5微升)在分析上具有挑战性。在这里，将开发和评估两种分析技术，用于非靶向代谢物分析；(1)液体萃取表面分析(LESA)与质谱仪联用，用于快速和自动化的样品提取和分析；(2)毛细管液相色谱-质谱法，用于分析脂肪和亲水性代谢物类别。OUTPUT-适用于干血和尿斑的非靶标代谢物分析方法。目的2-应用AIM 1开发的方法研究现有采集材料上干血和尿斑的稳定性；代谢物稳定性是这些研究中关注的问题。在这里，在一系列现有的文件和卡片上收集的干血和尿斑的稳定性将进行长达18个月的评估。同时，将开发和研究涂有酶和蛋白酶抑制剂的新的收集系统，以评估其适用性。这两项研究都将评估可检测到的代谢物类别的稳定性和多样性。OUTPUT-干血和尿点采集的方案和设备。目的3-应用(1)和(2)中开发的方法研究成年人久坐行为和运动中的新陈代谢。与伯明翰大学健康老龄化和锻炼的合作者将对应用DBS/DUS的成年人的久坐和锻炼行为进行纵向代谢研究。OUTPUT-关于不同年龄段久坐和锻炼受试者代谢变化的机械信息。

BACKGROUND AND SCIENTIFIC PROBLEM - The lifespan of the UK population is increasing at a greater rate than healthspan providing longer periods of ill health in later life and the associated economic strain on the NHS. There is a necessity to improve the healthy ageing of the UK population. The role of sedentary behaviour vs. exercise is being shown to have a strong influence on the healthy ageing-metabolism axis; exercise is a positive lifestyle intervention available to all. Metabolic changes can be studied applying nontargeted metabolomic analysis of thousands of metabolites. In these studies, biofluid samples are collected in a specialised centre which requires the subject to travel and expert staff and instruments to collect and process the samples; this process is costly and inefficient. However, sample collection away from specialised centres will reduce sample collection costs and allow longitudional studies to be performed cheaply and at minimal inconvenience to the subjects. The collection of dried blood or urine spots (DBS/DUS) away from specialised centres followed by metabolomic analysis offers a simple and robust solution. These devices are currently applied for targeted metabolite assays and we wish to develop this technology for non-targeted metabolomic studies of healthy ageing.SCIENTIFIC OBJECTIVE - We propose the development and application of dried blood and urine spot collection with non-targeted metabolomics to be applied in the study of healthy ageing.AIM 1 - Develop analytical methodologies for non-targeted metabolomic analysis of dried blood and urine spots. The small sample volumes (typically < 5microL) are analytically challenging in relation to sensitivity and range of metabolite classes detected. Here, two analytical techniques will be developed and assessed for non-targeted metabolomic analysis; (1) Liquid Extraction Surface Analysis (LESA) coupled to mass spectrometry for rapid and automated sample extraction and analysis and (2) capillary liquidchromatography-mass spectrometry for analysis of lipid and hydrophilic classes of metabolites.OUTPUT - analytical methods applicable for the non-targeted metabolomic analysis of dried blood and urine spots.AIM 2 - Study the stability of dried blood and urine spots on currently available collection materials applying methods developed in aim 1; the metabolite stability is a concern in these studies. Here, the stability of dried blood and urine spots collected on a range of currently available papers and cards will be assessed for up to 18 months. At the same time new collection systems coated with enzyme and protease inhibitors will be developed and studied to assess their applicability. Both of these studies will assess stability and diversity of metabolite classes detectable.OUTPUT - protocol and device for dried blood and urine spot collection.AIM 3 - Apply the methods developed in (1) and (2) to study metabolism in sedentary behaviour and exercise in adults. With collaborators in healthy ageing and exercise at the University of Birmingham a longitudional metabolomic study of sedentary vs. exercise behaviour in adults applying DBS/DUS will be performed.OUTPUT - mechanistic information on changes in metabolism in sedentary vs. exercising subjects atdifferent ages.