Development of Mass Spectrometric Techniques for Comprehensive Metabolome Analysis in Metabolomics

我的研究项目侧重于开发蛋白质组学和代谢组学的新质谱技术。与蛋白质组学相比，代谢组学是一个相对较新的领域，但有可能在研究系统生物学和寻找疾病生物标志物方面产生丰富的信息，通常是蛋白质组学的补充。代谢组学涉及对生物样本中存在的所有小分子代谢物（代谢物组）的研究。小分子是重要的细胞成分，可作为疾病生物标志物、表型指标和功能基因组学研究中的分子靶点。虽然近年来与代谢组学相关的出版物数量有所增加，但该领域仍然面临许多分析挑战，特别是在生成全面和定量代谢组学概况的领域；目前的分析工具只能检测和量化代谢组的一小部分。因此，我们提出的研究目标是开发新的质谱技术来解决这些分析挑战。

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.