Untargeted Metabolite Screening of Ewing Sarcoma Cells and Tissues with Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS).

Altered metabolism is a hallmark of cancer, where rewiring of cellular metabolic pathways is essential for tumor initiation, support of rapid proliferation, and adaptation to new metastatic sites. Metabolomics has emerged as a powerful tool to comprehensively characterize the metabolome of cancers and holds promise to transform understanding of how oncogenes can drive tumorigenesis and metastasis, identify cancer-specific metabolic dependencies, and provide novel therapeutic opportunities. This chapter describes methods to extract metabolites from Ewing sarcoma (EwS) cells grown in 2D or 3D cultures and from EwS mouse xenograft tissues, and to broadly analyze these compounds using an untargeted metabolite profiling approach. This methodology is likely applicable to a wide range of cultured cell and tissue types.