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Pattern recognition receptors link metabolite and bacteriaderived inflammation to insulin resistance during obesity. We demonstrate that NOD2 detection of bacterial cell wall peptidoglycan (PGN) regulates metabolic inflammation and insulin sensitivity. An obesitypromoting highfat diet (HFD) increased NOD2 in hepatocytes and adipocytes, and NOD2^−/− mice have increased adipose tissue and liver inflammation and exacerbated insulin resistance during a HFD. This effect is independent of altered adiposity or NOD2 in hematopoieticderived immune cells. Instead, increased metabolic inflammation and insulin resistance in NOD2^−/− mice is associated with increased commensal bacterial translocation from the gut into adipose tissue and liver. An intact PGNNOD2 sensing system regulated gut mucosal bacterial colonization and a metabolic tissue dysbiosis that is a potential trigger for increased metabolic …