DOP074Cell type–specific immunometabolic reprogramming in Crohn’s ileitis

Abstract Background Metabolic rewiring is integral to immune cell activation and tissue inflammation, but its spatial and cell type–specific organization in Crohn’s disease (CD) remains poorly understood. In this study, we aimed to construct a spatially resolved single-cell atlas of immunometabolic states in human ileal CD, spanning active inflammation, remission, and health, to identify activity-linked and persistent metabolic signatures. Methods Terminal ileal tissue was obtained from patients with active CD (n = 10), CD in remission (n = 8), and controls (n = 10). Imaging mass cytometry was performed using a 37-marker panel incorporating lineage, structural and 14 metabolic proteins. Data were analyzed at single-cell resolution to define metabolic programs across identified cell clusters and spatial neighborhoods. In parallel, peripheral blood mononuclear cells were profiled by suspension mass cytometry to assess systemic signatures. Comparative and spatial correlation analyses were conducted in R using Phenograph clustering and neighborhood interaction modelling. Results IMC revealed extensive, cell type–resolved metabolic reprogramming in active CD. Inflammation was characterized by reduced mitochondrial and tricarboxylic acid cycle activity (ATP5A, CytC, VDAC1) with a concurrent shift towards the pentose phosphate pathway (G6PD) and dampened mTOR signaling. CD68+ macrophages and neutrophils displayed pronounced PPP activation, whereas B cells showed persistent depression of anaplerosis (GLUD1/2) and lipid uptake (CD36). Spatial neighborhood analysis uncovered compartment-specific metabolic coupling, particularly in remission, where immune-epithelial proximity correlated with restored mitochondrial activity. Peripheral blood exhibited only modest, myeloid-restricted changes, indicating that metabolic rewiring is predominantly tissue-embedded. Conclusion Crohn’s ileitis is defined by spatially organized, cell type–specific metabolic remodeling that persists beyond clinical remission. This immunometabolic atlas provides a framework for identifying tissue-specific therapeutic targets and biomarkers of disease activity in inflammatory bowel disease. Conflict of interest: Lehmann, Malte: No conflict of interest Walling, Sonja: No conflict of interest Adrian, Huck: No conflict of interest Kunkel, Désirée: No conflict of interest Glauben, Rainer: No conflict of interest Kühl, Anja A.: No conflict of interest Siegmund, Britta: Grant: Pfizer Other: Consultant: Abbvie, Abivax, Boehringer Ingelheim, Bristol Myers Squibb, Dr. Falk Pharma, Eli Lilly, Endpoint Health, Galapagos, Janssen/Johnson & Johnson, Materia Prima, MSD, Pfizer, Takeda, Wedbush Securities. Speaker: Abbvie, AlfaSigma, Bristol Myers Squibb, CED Service GmbH, Dr. Falk Pharma, Eli Lilly, Ferring, Galapagos, Janssen/Johnson & ampJohnson, MD Education, MSD, Pfizer, Tr1xBio. Haag, Lea Maxie: No conflict of interest