High Throughput Lipidomic Profiling with High Structural Specificity for Phenotypic Screening - PROJECT ABSTRACT Lipids are a diverse class of biomolecules that play pivotal roles in various cellular processes and have been implicated in the pathogenesis of diseases such as diabetes, Alzheimer's, Parkinson's, and various cancers. Understanding lipid structures and functions is crucial for unraveling their roles in complex biological systems and diseases. Unambiguous identification of lipids, particularly with high-level structural details and separation of lipid isomers, has been an analytical challenge. However, such knowledge is essential for obtaining a full understanding of lipid functions and discovering lipid biomarkers for diseases. Addressing the absence of rapid lipid profiling methods and that provide sufficient structural details is critical for predicting, controlling, and engineering lipid metabolism on a large scale for biomedical and phenotypic screening. My long-term goal is to enable the elucidation of the detailed molecular mechanisms of lipid metabolism through advancing lipidomic profiling with high structural specificity and to unravel the relationship between specific lipids and associated functions in biological systems and diseases. To achieve this, we propose developing a next-generation lipidomics pipeline using mass spectrometry-based technologies and advanced bioinformatics capabilities. Our approach involves developing a multi-dimensional lipidomic platform using high-resolution structures for lossless ion manipulations-based ion mobility spectrometry-mass spectrometry, advanced fragmentation techniques, and gas-phase reactions for effective lipid structure elucidation, including resolved isomer separation and determining double bond locations. This integrative platform will be coupled with liquid chromatography and a novel data acquisition approach utilizing both data-dependent acquisition and data- independent acquisition for a comprehensive analysis of lipids in complex samples. To ensure robust data processing, lipid identification and structure elucidation, we will develop advanced bioinformatics pipelines using artificial intelligence approaches. Additionally, a high-throughput and quantitative workflow utilizing online solid-phase extraction for rapid lipid screening will be developed. The robustness of this workflow and advanced bioinformatics pipeline will be validated by fully profiling the yeast lipidome. A comprehensive and structurally detailed lipid atlas for the yeast knockout collection will be constructed and provide a foundation for understanding lipid metabolism and associated functions analogous to human genes. We anticipate that this platform will become a fundamental capability for lipid profiling in biomedical research to unravel the intricate relationship between lipids and associated functions in biological systems and human health. Furthermore, we expect this innovative approach to facilitate the discovery of lipid biomarkers for disease diagnostics and drug development.
