High Resolution Mass Spectrometry to enhance hormonal profiling across the lifespan.

高分辨率质谱分析可增强整个生命周期的荷尔蒙分析。

• 批准号: BB/V019066/1
• 负责人: Ruth Andrew
• 金额: $ 95.44万
• 依托单位: University of Edinburgh
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FV019066%2F1
• 关键词: Resolution; Mass; Spectrometry; enhance; hormonal

Hormones formed from cholesterol, known as "steroids", play critical roles in regulating stress, inflammation, metabolism, muscle and bone strength and fertility. Understanding how hormones control body functions is key to supporting life-long health, e.g. do hormones during pregnancy determine wellbeing during life and do they determine physical and mental frailty with age? Our team investigate nutrition and stress during life-stages e.g. pregnancy, childhood, menopause, ageing and study the differences in health of women vs men with age, e.g. protection against heart and lung disease and dementia. To gain insight into hormonal signalling, researchers must assess a wide array of molecules from large ones (like DNA and proteins) to small molecules, known as "metabolites", each with their own role to play. To measure small metabolites, we use machines called "mass spectrometers", which weigh molecules and can identify them. In the Edinburgh Mass Spectrometry Core, we focus on "targeted hormonal profiles" to allow researchers to investigate amounts and turnover of sets of closely related hormones in humans and animal models. Researchers interrogate hormone families of interest and our particular expertise lies in profiling steroid hormones. "Targeted profiling" of all members of the steroid hormone family, in excess of 40, offers a road map of how these hormones are activated and inactivated, in other words a "fingerprint" of health status. When a particular hormonal pathway is earmarked as vital to a body response, we can measure the building bricks used to create the hormones, the amounts of active hormones and the breakdown products all together, letting us know how quickly the body is making and removing essential hormones. We have gradually expanded the technology in our lab to encompass a hand-picked suite of instruments well matched to targeted hormonal profiling. We have refined pertinent hormonal screens and adapted measurements in adult humans to smaller volume samples to allow us to study babies and animal models with the same fidelity. However, in widening our view, we have rendered the profiles increasingly complex, bringing with it challenges. It is becoming harder for us to be sure we are not inadvertently measuring two or more closely related hormones at the same time by mistake. We can use a technique called "chromatography" to spread out the steroids to limit interference. Indeed we are skilled at this, but as complexity rises this approach causes longer analysis times and slows down lab productivity. In the current era of large population studies, we must achieve highly specific readouts but with faster throughput. Therefore we are applying to install a new "high resolution" mass spectrometer to support high-turnaround and high quality hormonal profiling. This system will allow us to measure weights of hormones more accurately so we can confidently work in a faster timeframe. We will also upgrade to include automated sample preparation. We currently inject a sample into the instrument, perform our analysis and then wait while the system stabilises before moving onto the next sample. With the new instrument we will operate two injectors in tandem, which will allow us to switch between them and thus run twice as fast. One will operate while the other stabilises and resets and vice versa. We will be able to measure more complex mixtures in single experiments, faster. This will be a big step forward from our currently set-up where we have to analyse the sample several times, often consuming more sample than is routinely available from infants and animals. We will initially roll-out projects related to early-life stress in babies, and effects of gender on lung health and growth in humans and animals, including mice, sheep and quail. We will make the approaches we develop available to support many University researchers and industries and we will communicate our findings quickly to scientists and the public.

由胆固醇形成的激素，被称为“类固醇”，在调节压力、炎症、新陈代谢、肌肉和骨骼强度以及生育能力方面起着关键作用。了解激素如何控制身体功能是支持终身健康的关键，例如，怀孕期间的激素是否决定了一生的幸福，它们是否决定了随着年龄的增长而出现的身体和精神脆弱？我们的团队调查了生命阶段的营养和压力，例如怀孕，儿童，更年期，衰老，并研究了女性与男性随着年龄增长的健康差异，例如预防心脏和肺部疾病以及痴呆症。为了深入了解激素信号，研究人员必须评估从大分子（如DNA和蛋白质）到小分子（称为“代谢物”）的各种分子，每种分子都有自己的作用。为了测量小的代谢物，我们使用称为“质谱仪”的机器，它可以称量分子并识别它们。在爱丁堡质谱核心中，我们专注于“靶向激素谱”，以使研究人员能够调查人类和动物模型中密切相关的激素的数量和周转率。研究人员询问感兴趣的激素家族，我们的特殊专长在于分析类固醇激素。对超过40种类固醇激素家族的所有成员进行“有针对性的分析”，提供了这些激素如何被激活和失活的路线图，换句话说，提供了健康状况的“指纹”。当一个特定的激素途径被指定为对身体反应至关重要时，我们可以测量用于产生激素的积木，活性激素和分解产物的数量，让我们知道身体制造和去除必需激素的速度。我们已经逐渐扩大了我们实验室的技术，包括一套精心挑选的仪器，与有针对性的激素分析相匹配。我们已经完善了相关的激素筛选，并将成年人的测量调整为较小体积的样本，使我们能够以相同的保真度研究婴儿和动物模型。然而，在扩大我们的视野时，我们使概况变得越来越复杂，带来了挑战。我们越来越难以确定我们是否无意中错误地同时测量了两种或更多种密切相关的激素。我们可以用一种叫做“色谱法”的技术来分散类固醇以限制干扰。事实上，我们在这方面很熟练，但随着复杂性的增加，这种方法会导致更长的分析时间，并降低实验室的生产力。在当前的大人群研究时代，我们必须实现高度特异性的读数，但要有更快的吞吐量。因此，我们正在申请安装一台新的“高分辨率”质谱仪，以支持高周转和高质量的激素分析。这个系统将使我们能够更准确地测量激素的重量，这样我们就可以在更快的时间内自信地工作。我们还将进行升级，以包括自动化样品制备。我们目前将样品注入仪器，进行分析，然后等待系统稳定，然后再进入下一个样品。有了新仪器，我们将操作两个注射器串联，这将使我们能够在它们之间切换，从而运行速度提高一倍。一个将运行，而另一个稳定和重置，反之亦然。我们将能够在单个实验中更快地测量更复杂的混合物。这将是一个很大的进步，从我们目前的设置，我们必须分析样品几次，往往消耗更多的样品比常规提供的婴儿和动物。我们将首先推出与婴儿早期生活压力有关的项目，以及性别对人类和动物（包括小鼠，绵羊和鹌鹑）肺部健康和生长的影响。我们将使我们开发的方法可用于支持许多大学的研究人员和行业，我们将迅速向科学家和公众传达我们的发现。