Calcium/calmodulin-dependent protein kinase kinase 2 regulates hepatic fuel metabolism

The liver is the primary internal metabolic organ that coordinates whole body energy homeostasis in response to feeding and fasting. Genetic ablation or pharmacological inhibition of calcium/calmodulin-dependent protein kinase kinase 2 (CaMKK2) has been shown to significantly improve hepatic health and peripheral insulin sensitivity upon overnutrition with high fat diet. However, the precise molecular underpinnings that explain this metabolic protection have remained largely undefined. To characterize the role of CaMKK2 in hepatic metabolism, we developed and challenged liver-specific CaMKK2 knockout (CaMKK2LKO) mice with high fat diet and performed glucose and insulin tolerance tests to evaluate peripheral insulin sensitivity. We used a combination of RNA-Sequencing, glucose and fatty acid istotopic tracer studies, a newly developed Seahorse assay for measuring the oxidative capacity of purified peroxisomes, and a degenerate peptide libarary to identify putative CaMKK2 substrates that mechanistically explain the protective effects of hepatic CaMKK2 ablation. Consistent with previous findings, we show that hepatic CaMKK2 ablation significantly improves indices of peripheral insulin sensitivity. Mechanistically, we found that CaMKK2 phosphorylates and regulates GAPDH to promote glucose metabolism and PEX3 to blunt peroxisomal fatty acid catabolism in the liver. CaMKK2 is a central metabolic fuel sensor in the liver that significantly contributes to whole body systems metabolism. •Hepatic CaMKK2 is sufficient to toggle peripheral insulin sensitivity.•CaMKK2 is required for hepatic glucose metabolism.•Hepatic CaMKK2 balances glycolytic and fatty acid metabolism.•CaMKK2 promotes glycolysis through regulation of GAPDH.•CaMKK2 suppresses fatty acid metabolism through Pex3.