Development of Mass Spectrometric Techniques for Comprehensive Metabolome Analysis in Metabolomics

我的研究计划专注于为蛋白质组学和代谢组学开发新的质谱学技术。与蛋白质组学相比，代谢组学是一个相对较新的领域，但在研究系统生物学和寻找疾病生物标志物方面，代谢组学具有产生丰富信息的潜力，通常是对蛋白质组学的补充。代谢组学涉及对生物样本中存在的所有小分子代谢物(代谢组)的研究。小分子是重要的细胞成分，在功能基因组研究中可以作为疾病的生物标志物、表型指示物和分子靶标。虽然近年来与代谢组学有关的出版物数量有所增加，但该领域仍然面临一些分析挑战，特别是在生成全面和定量的代谢组图谱方面；目前的分析工具只能检测和量化一小部分代谢组。因此，我们提出的研究的目标是开发新的质谱学技术来解决这些分析挑战。 为了实现我们的目标，我们将开发一个高代谢组覆盖率、高检测特异性和高通量的定量代谢组谱的综合分析平台。该平台的工作流程将包括代谢物样品的前处理、代谢物提取、代谢物的同位素标记、标记代谢物的液相色谱(LC)分离结合质谱仪(MS)和串联质谱仪(MS/MS)用于代谢物的定量和鉴定，以及数据分析。我们将开发新的方法来提高此工作流中每个步骤的分析性能。具体地说，我们将开发(1)通过有效地分离化学基团来处理代谢组样本的改进的技术，(2)针对不同代谢物或次代谢组的新的和改进的同位素标记化学，以增加整体代谢组的覆盖率，(3)多维分离工具，以降低代谢组样本的复杂性，用于改进的MS分析，(4)新的和改进的MS和MS/MS策略，以提高检测的灵敏度和特异度，(5)扩展的代谢组库，包括实验和预测的MS/MS谱，以及(6)基于网络的化合物鉴定公共资源。在开发阶段，我们将把每种开发技术应用于真实世界的样本，以衡量其分析性能，并说明该方法的实用价值。我们还将调查每种方法操作背后的物理和化学过程，以进一步优化其分析性能。最终，我们将整合这些方法来开发一个分析平台，该平台将能够高效和特异地生成全面和定量的生物样本代谢组谱。 这项研究的新颖性和预期意义在于开发了一种新的基于MS的分析技术，可用于大规模的生物系统研究，如发现用于疾病诊断或预后的特定生物标记物，监测个体的健康状况以改善生活方式并及早发现与身体健康和心理健康相关的疾病，以及应用系统生物学从整体上调查和了解生物系统的功能。由于我们预计代谢组学在未来的许多领域将发挥越来越重要的作用，这一研究计划为培养一批高技能和极受欢迎的分析化学家提供了巨大的机会。

My research program focuses on developing new mass spectrometric techniques for proteomics and metabolomics. Compared to proteomics, metabolomics is a relatively new field, but has the potential to produce a wealthy array of information, often complementing proteomics, in studying systems biology and searching for disease biomarkers. Metabolomics involves the study of all the small molecule metabolites (the metabolome) present in a biological sample. Small molecules are important cellular components that can be used as disease biomarkers, as phenotype indicators and as molecular targets in functional genomics research. While the number of publications related to metabolomics has increased in recent years, this field still faces a number of analytical challenges, particularly in the area of generating comprehensive and quantitative metabolome profiles; current analytical tools can detect and quantify only a small fraction of the metabolome. Thus the objective of our proposed research is to develop new mass spectrometric techniques to address some of these analytical challenges. To achieve our goal, we will develop an integrated analytical platform for quantitative metabolome profiling with high metabolome coverage, high detection specificity and high throughput. The workflow of this platform will involve several steps, including preprocessing of metabolomic samples, metabolite extraction, isotope labeling of metabolites, liquid chromatography (LC) separation of the labeled metabolites combined with mass spectrometry (MS) and tandem MS (MS/MS) for metabolite quantification and identification, and data analysis. We will develop new methods to improve the analytical performance of each step in this workflow. Specifically, we will develop (1) improved techniques to process the metabolome samples with efficient fractionation of chemical groups, (2) new and improved isotope labeling chemistries to target different groups of metabolites or sub-metabolomes in order to increase the overall metabolome coverage, (3) multidimensional separation tools to reduce metabolome sample complexity for improved MS analysis, (4) new and improved MS and MS/MS strategies to increase detection sensitivity and specificity, (5) an expanded metabolome library including experimental and predicted MS/MS spectra, and (6) a web-based public resource for compound identification. During the development stage, we will apply each developing technique to real world samples to gauge its analytical performance and illustrate the practical utility of the method. We will also investigate the physical and chemical processes underlying the operation of each method so as to further optimize its analytical performance. In the end, we will integrate these methods to develop an analytical platform that will enable the generation of comprehensive and quantitative metabolome profiles of biological samples with high efficiency and specificity. The novelty and expected significance of this research lies in the development of a new MS-based analytical technology that can be used for large scale studies of biological systems, such as discovery of specific biomarkers for diagnosis or prognosis of diseases, monitoring the health status of individuals leading to better life style and early detection of disorders related to physical health and psychological well-being, and applying systems biology to investigate and understand the functions of a biosystem holistically. As we anticipate that metabolomics will play an increasingly important role in many areas in the future, this research program provides great opportunities for training a number of highly skilled and highly sought-after analytical chemists.