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Background Intestinal epithelial barrier disruption and immune microenvironment chaos are hallmarks of inflammatory bowel disease (IBD). While mature intestinal epithelial cells (IECs) are known to undergo a glycolytic shift under inflammatory stress, the mechanisms by which this metabolic reprogramming executes epithelial death remained understood. This study aimed to investigate whether glycolytic metabolites triggered membrane lysis and orchestrated inflammatory.Methods Primary IECs and 3D intestinal organoids were subjected to metabolic stress. Protein pyruvylation was identified and quantified using LC-MS/MS combined with pan-pyruvylation antibodies. Spatiotemporal dynamics of mitochondria-plasma membrane contacts were visualized. Experimental colitis models were performed using IEC-specific Rictor K-to-R mutation and neutrophil-specific Tlr9/Sting depletion.Results We found that the dysbiotic IBD microenvironment induced intracellular pyruvate accumulation in mature IECs. This metabolite drove pyruvylation on the mTORC2 complex and caused mTORC2 pathological hyperactivation. Hyperactive mTORC2 suppressed F-actin cytoskeletal dynamics, leading to the spatial de-tethering of ROS-rich, damaged mitochondria. Combined with disrupted autophagy via ROS-oxidized ULK1, these unanchored mitochondria migrated to the plasma membrane and executed localized lipid peroxidation, resulting in cell lysis. The liberated mtDNA activated the neutrophil TLR9/cGAS-STING axis and triggered NETosis. This cascade degraded mucus layers and tight junctions in colitis mice.Conclusions This study presented the metabolism-space-immune pathogenic axis in IBD ( IDDF2026-ABS-0544 Figure 1. Visual abstract). Our findings demonstrate that glycolytic pyruvylation acts as a critical metabolic switch in mitochondrial spatial toxicity. Targeting the axis represents a promising therapeutic strategy for restoring intestinal barrier integrity.Abstract IDDF2026-ABS-0544 Figure 1