Spatial metabolomics strategy reveals heterogeneity of prostate cancer based on multi-platform imaging and laser microdissection-gas chromatography-tandem mass spectrometry.

Prostate cancer (PCa) exhibits significant intra-tumor metabolic heterogeneity. In this study, we developed a multi-platform workflow combining pathological staining, matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI), and laser capture microdissection-assistant gas chromatography-tandem mass spectrometry (LCM-GC-MS/MS) to investigate the metabolic spatial heterogeneity of a specific PCa tissue. Four suspicious glandular regions were initially identified by histology and immunofluorescence. MALDI-MSI revealed three distinct metabolic patterns for further analysis. It was found that only one glandular region showed PCa metabolic features, enabling precise differentiation from other regions of interest. The information from LCM-GC-MS/MS complemented that from MSI by confirming and quantifying fatty acid alterations. This workflow broadened the spatial metabolic coverage and enhanced intra-tumor metabolic heterogeneity analysis in PCa.