Spatial metabolomics reveals region-specific metabolic alterations in progressive multiple sclerosis

Progressive multiple sclerosis (P-MS) represents a major challenge for neuroscience research due to the lack of an effective therapeutic strategy and its complex pathogenesis. Conventionally, P-MS is known to be characterized by pronounced neuroinflammatory processes associated with demyelination and neurodegeneration. P-MS patients also show marked metabolic alterations occurring in both immune and neuronal cells. Nevertheless, the connection between metabolic impairment and disease progression remains poorly investigated field. To fill this knowledge gap, we used chronical experimental autoimmune encephalomyelitis (cEAE), which recapitulates key pathological features of P-MS, to investigate metabolic impairments by spatial mass spectrometry. Moreover, we decided to administer a metabolic reprogramming therapy in these animals at chronic stage, in order to correct the metabolic alteration observed and rescue the pathological phenotype. Spatial mass spectrometry analyses enabled high-resolution mapping of metabolic alterations within the spinal cord among wild type, cEAE, and treated groups. UMAP-based clustering identified distinct metabolic signatures for each condition, revealing disease-associated metabolic disorganization and partial recovery upon treatment. Moreover, K-means clustering allowed the successful discrimination of white and grey matter regions within the spinal cord and spatial clustering analysis revealed a distinct metabolic cluster located in the white matter of WT mice that was lost in cEAE animals and only partially restored in the treated group. Enrichment and spatial correlation analyses revealed major alterations in alanine, aspartate, and glutamate metabolism in EAE mice, pathways tightly linked to glutamate excitotoxicity and oxidative stress. In addition, significant upregulation of arachidonic acid and unsaturated fatty acid biosynthesis was observed, suggesting ongoing inflammatory activity and membrane remodeling. Treatment partially restored amino acid metabolism toward WT-like levels, while lipid metabolism showed only partial recovery, consistent with incomplete resolution of inflammation and lipid signaling disruption. Together, these results provide new insights into the metabolic mechanisms underlying disease progression and suggest that targeting metabolic pathways may represent a novel strategy to mitigate chronic neuroinflammation and neurodegenerative damage in progressive multiple sclerosis.