Unveiling spatial metabolic alterations driving disease progression in progressive multiple sclerosis

Progressive multiple sclerosis (P-MS) represents a major challenge due to the complexity of its pathogenesis and the lack of effective therapies. P-MS is characterized by chronic neuroinflammation, demyelination, neurodegeneration, and profound metabolic alterations in both neuronal and immune cells. However, the contribution of these metabolic deficits to disease progression remains poorly understood. To address this gap, we employed the chronic experimental autoimmune encephalomyelitis (cEAE) mouse model, which recapitulates key features of P-MS, and applied high resolution spatial mass spectrometry (SMS) to map metabolic alterations across brain regions. This multimodal approach integrates spatial metabolomics with computational clustering and dimensionality reduction analyses. UMAP clustering revealed distinct metabolic profiles in wild type and cEAE mice, indicating pronounced metabolic disorganization in cEAE. Spatial cluster analysis delineated white and grey matter regions and identified a white matter specific metabolic cluster that was lost in cEAE. Enrichment and spatial correlation analyses revealed marked alterations in alanine, aspartate, and glutamate metabolism pathways linked to the Krebs cycle, excitotoxicity, and oxidative stress together with increased biosynthesis of arachidonic acid and unsaturated fatty acids, consistent with persistent inflammatory signaling. To counteract these alterations, we tested the effect of metabolic reprogramming therapy in cEAE mice based on the administration of metabolic precursors, including essential amino acids, Krebs cycle intermediates, and co-factors. SMS analysis showed that treated mice exhibited an intermediate metabolic phenotype between wild type and cEAE animals, with partial restoration of amino acid and lipid related metabolic pathways. Overall, this study demonstrates that spatially resolved metabolic mapping combined with computational analyses can help clarify mechanisms underlying disease progression in P MS. These findings support targeted metabolic modulation as a promising approach to restore neuronal function and reduce chronic neuroinflammation and neurodegeneration.