Spatial and Untargeted Metabolomics Define Location- and Histology-Specific Metabolic Reprogramming in Colorectal Cancer

Colorectal cancer (CRC) exhibits profound molecular heterogeneity based on primary tumor location. A comprehensive understanding of the spatially resolved metabolic differences between right-sided (RCC) and left-sided (LCC) carcinomas remains elusive. Here, to eliminate the influence of inter-individual heterogeneity on metabolic differences, we performed spatial metabolomics using matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) on synchronous bilateral tumors from the same patient. This was complemented by untargeted metabolomic profiling via ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) on tumor and matched normal tissues from 30 CRC patients with different locations and histologies. Our analysis identified 345 metabolites and revealed distinct, location-specific metabolic reprogramming. Among these, 72 altered metabolites between left- (Ltu) and right-sided colon tubular adenocarcinomas (Rtu) indicated disruptions in ether lipid and phosphatidylcholine metabolism of Ltu and nucleotide, amino sugar, and fatty acid metabolism of Rtu. Furthermore, right-sided mucinous adenocarcinomas (Rmu) were characterized by unique sphingolipid metabolism reprogramming, shifting from an 'apoptosis-promoting signaling' to a 'structural construction' function, compared to Rtu. N-Acetylneuraminic acid, oleoyl-l-carnitine and 12-Hydroxystearic acid are representative DEMs in the aforementioned pathway. Spatial analysis delineated metabolic profiles varied spatially from the tumor center to the periphery. Collectively, our findings underscore the influence of primary tumor location on metabolism and suggest subtype-specific metabolic adaptation mechanisms associated with histology and spatial architecture.