Precision-cut liver slices as an ex vivo model to assess impaired hepatic glucose production

Abstract Fasting hypoglycemia is a severe and incompletely understood symptom of various inborn errors of metabolism (IEM). Precision-cut liver slices (PCLS) represent a promising model to study glucose production ex vivo. This study aimed to provide a comprehensive characterization of glucose production in human and murine PCLS. We quantified net glucose production rate of PCLS derived in the presence of commonly used gluconeogenic precursors and from fed and fasted mice. Dihydroxyacetone (DHA) yielded the highest rate, which was further stimulated by forskolin and dibutyryl-cAMP. Moreover, using 13C isotope tracing, we assessed the contribution of glycogenolysis and gluconeogenesis to net glucose production over time, showing faster incorporation of DHA than glycerol in the gluconeogenesis. Pharmacological inhibition of the glucose 6-phosphate transporter SLC37A4 markedly reduced net glucose production and increased lactate secretion and glycogen storage, while glucose production was completely abolished in PCLS from glycogen storage disease (GSD) type Ia and Ib patients. This indicates that PCLS allow to model the primary clinical symptoms of GSD type I, an inborn error of carbohydrate metabolism. In conclusion, this study identifies PCLS as an effective ex vivo model to study hepatic glucose production and opens opportunities for its future application in IEM research and beyond.