Building research capacity with high-throughput Ultra-performance liquid chromatography high resolution Q-TOF

利用高通量超高效液相色谱高分辨率 Q-TOF 建设研究能力

• 批准号: MR/X013537/1
• 负责人: Zoltan Takats
• 金额: $ 85.8万
• 依托单位: Imperial College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FX013537%2F1
• 关键词: Building; research; capacity; throughput; Ultra

The analysis of biological molecules by mass spectrometry (LC-MS) enables the identification and quantification of many thousands of metabolites, lipids, and proteins in a single measurement. Over the last 10-15 years, there has been a revolution in instrument sensitivity and resolution allowing deeper insight into the molecular processes inside living cells. At Imperial College London, we have world-leading capabilities for metabolite and lipid characterisation from clinical and research samples and have pioneered workflows to image the molecular footprint of clinical tissue (such as cancer biopsies). However, we lack the technical instrumentation for similar proteomics workflows. We wish to establish a state-of-the-art multi-user platform to support health research with cutting-edge proteomics applications and small-molecule detection. Investment in a new LC-MS platform will enable the unique capacity for comprehensive molecular footprinting of clinical samples, leading to a greater understanding of the chemical reactions that give us life, and new ways of treating disease. The instrument that we have identified (EvoSep - 7600 ZenoTOF) has unique attributes to allow us to identify different classes of molecules (such as bile-acid species) and analyse thousands of proteins in a high-throughput manner (~20 minutes per sample) to support research in key areas such as neurodegeneration, cancer, nutrition, experimental medicine and anti-microbials.Neurodegeneration: The Dementia Research Institute in Imperial College have hundreds of blood and brain samples for studying diseases such as Alzheimer's disease (AD). In particular, understanding how different genetic variants can lead to AD will be determined, in part, through proteomic analysis of >260 brains, leading to greater insight into this currently incurable disease. Other areas of priority areas include Huntington's disease and chronic pain.Cancer: Imaging mass spectrometry of cancer tissue has been pioneered by our research group, revealing distinct regions within tumours that may cause different responses to chemotherapies. We can use a laser to dissect these distinct regions (sometimes only a few cells) and further characterise the molecular pathways by highly sensitive proteomics and metabolomic profiling. We wish to apply this to better understand cancers and their response to drugs, in particular breast and glioblastoma.Nutrition and experimental medicine: Detecting changes in nutrient availability is essential to maintaining healthy physiology. Understanding nutrient-sensing requires detecting distinct classes of molecules (bile acids and signalling peptides) at low abundance in biological fluids. The sensitivity of the 7600 ZenoTOF will permit quantification of the low abundance of signalling peptides within blood while the unique EAD fragmentation feature will characterise bile acids to identify new relationships with gut microbiota. Gut microbiotas play a fundamental role in gastrointestinal health, this platform will contribute to understanding the interaction between gut microbiota and ourselves as well as the mechanisms behind the health benefits of intestinal microbiota transplantation.Antimicrobial resistance: The recent pandemic has highlighted the urgent need for new antivirals. Using LC-MS, we will investigate the relationship between viruses and their host (humans) by applying technologies that "label" proteins coming into contact with the viral genetic material. In this way, we detect what the virus interacts with at a molecular level to replicate itself. Understanding this can lead to the discovery of new ways to treat viruses, by producing antivirals that disrupt these interactions. It will also support research into the development of a new non-antibiotic class of anti-microbials that disrupt how bacteria pathologically interact with our body.

通过质谱法（LC-MS）分析生物分子使得能够在单次测量中鉴定和定量数千种代谢物、脂质和蛋白质。在过去的10-15年里，仪器灵敏度和分辨率发生了革命性的变化，使人们能够更深入地了解活细胞内的分子过程。在伦敦帝国理工学院，我们拥有世界领先的临床和研究样本代谢物和脂质表征能力，并开创了临床组织（如癌症活检）分子足迹成像的工作流程。然而，我们缺乏类似蛋白质组学工作流程的技术工具。我们希望建立一个最先进的多用户平台，以支持具有尖端蛋白质组学应用和小分子检测的健康研究。对新LC-MS平台的投资将使其具有对临床样本进行全面分子足迹分析的独特能力，从而更好地了解赋予我们生命的化学反应以及治疗疾病的新方法。我们发现的仪器（EvoSep - 7600 ZenoTOF）具有独特的属性，使我们能够识别不同类别的分子（如胆汁酸种类），并以高通量的方式分析数千种蛋白质（每个样本约20分钟），以支持关键领域的研究，如神经变性，癌症，营养，实验医学和抗微生物。帝国理工学院的痴呆症研究所拥有数百份血液和大脑样本，用于研究阿尔茨海默病（AD）等疾病。特别是，了解不同的遗传变异如何导致AD将部分通过对>260个大脑的蛋白质组学分析来确定，从而更深入地了解这种目前无法治愈的疾病。其他优先领域包括亨廷顿氏病和慢性疼痛。癌症：我们的研究小组开创了癌症组织的成像质谱，揭示了肿瘤内可能对化疗产生不同反应的不同区域。我们可以使用激光来解剖这些不同的区域（有时只有几个细胞），并通过高灵敏度的蛋白质组学和代谢组学分析进一步研究分子途径。我们希望将其应用于更好地了解癌症及其对药物的反应，特别是乳腺癌和胶质母细胞瘤。营养和实验医学：检测营养物质可用性的变化对维持健康的生理至关重要。了解营养传感需要检测不同类别的分子（胆汁酸和信号肽）在生物液体中的低丰度。7600 ZenoTOF的灵敏度将允许对血液中低丰度的信号肽进行定量，而独特的EAD片段化功能将使胆汁酸分解，以确定与肠道微生物群的新关系。肠道菌群在胃肠道健康中起着基础性的作用，该平台将有助于了解肠道菌群与我们自身之间的相互作用，以及肠道菌群移植对健康益处背后的机制。抗生素耐药性：最近的大流行凸显了对新抗病毒药物的迫切需求。使用LC-MS，我们将通过应用“标记”与病毒遗传物质接触的蛋白质的技术来研究病毒与其宿主（人类）之间的关系。通过这种方式，我们可以检测到病毒在分子水平上与什么相互作用以复制自己。了解这一点可以通过生产破坏这些相互作用的抗病毒药物来发现治疗病毒的新方法。它还将支持开发一种新的非抗生素类抗微生物药物的研究，这种药物可以破坏细菌与我们身体的病理相互作用。

项目成果

Intracellular Chloride Channels Regulate Endothelial Metabolic Reprogramming in Pulmonary Arterial Hypertension.

细胞内氯离子通道调节肺动脉高压的内皮代谢重编程。

• DOI: 10.1165/rcmb.2022-0111oc
• 发表时间: 2023
• 期刊: American journal of respiratory cell and molecular biology
• 影响因子: 6.4
• 作者: Alzaydi MM

An organ-on-chip model of pulmonary arterial hypertension identifies a BMPR2-SOX17-prostacyclin signalling axis.

• DOI: 10.1038/s42003-022-04169-z
• 发表时间: 2022-11-07
• 期刊: Communications biology
• 影响因子: 5.9