Pathology-Guided Spatial Metabolomics Characterizes Glycerophospholipid Remodeling Associated with Radioresistance in Nasopharyngeal Carcinoma

Radioresistance limits durable control of nasopharyngeal carcinoma (NPC), but pretreatment tumor-region metabolic patterns associated with subsequent radiotherapy response remain incompletely characterized. We applied pathology-guided matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) to pretreatment fresh-frozen NPC tissues from three radiosensitive (RS) and two radioresistant (RR) patients. Histologically annotated tumor regions of interest (ROIs) were registered to MSI data, and LC-MS/MS profiles from the corresponding residual tissue blocks supported molecular annotation. Independent proteomic and transcriptomic cohorts were used for cross-omics contextualization. Exploratory analysis of the pathology-defined tumor ROIs highlighted a glycerophospholipid-associated pattern involving phosphatidic acid, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, phosphatidylcholine, and phosphatidylglycerol species. Preranked pathway analysis further highlighted glycerophospholipid metabolism together with membrane- and stress-related processes. In the broader transcriptome- and proteome-driven network analyses, glycerophospholipid metabolism was the only shared human-relevant pathway that also appeared in the pathology-guided MSI enrichment. In this exploratory five-patient cohort, pooled-pixel analysis described a pathology-localized glycerophospholipid-associated pattern in the RR-versus-RS comparison. No MSI feature achieved patient-level significance after multiple-testing correction, and the direction of the highlighted lipid pattern was sensitive to aggregation. These findings are hypothesis-generating and warrant evaluation in larger, independently accrued cohorts.