(237b) Stiffness Induces Metabolic Dysfunction in Primary Hepatocytes

Liver is a complex organ that performs myriad functions as a center of metabolism, having key roles in synthesis, detoxification, and digestion. Liver stiffness has been established as a useful clinical metric to stage liver disease and in recent years as a prognostic tool predictive of outcomes. However, it remains unclear how tissue stiffness itself contributes to hepatic dysfunction. To study the effect of stiffness on liver function, we employed our BEASTS (Bio-Engineered Adhesive Siloxane Substrate with Tunable Stiffness) platform to recreate the range of stiffness observed in healthy to highly fibrotic environments in cell culture. A significant challenge in in vivo models, our approach allows for control of stiffness corresponding to various stages of fibrosis. Primary hepatocytes were cultured on BEASTS mimicking liver stiffness from healthy to highly fibrotic tissue. Key markers of hepatic function, urea and albumin production, decreased when hepatocytes were cultured on fibrotic stiffness. Enhanced glycolytic capacity and mitochondrial respiration were exhibited by hepatocytes cultured on fibrotic stiffness. Greater intracellular lipid accumulation was observed in hepatocytes cultured on fibrotic stiffness, along with dysregulated lipid metabolic genes. Hepatocytes cultured on fibrotic stiffness displayed a lower abundance and shift in the redox balance of intracellular glutathione, suggesting impairment of the oxidative stress response and a plausible mechanism for elevated oxidative stress in the fibrotic environment. Together our data suggest a role of increased liver stiffness in driving hepatocyte metabolic dysfunction and contributing to loss of organ function in the setting of liver disease. A more nuanced approach to therapeutic development for liver fibrosis can be aided by data from studying how tissue stiffness influences hepatocyte biology.