Affecting 25% of the world population, non-alcoholic fatty liver disease (NAFLD) is becoming the leading cause of liver failure and liver cancers in the 2040s. Annually, 0.3-2.2% of NAFLD patients progress from benign steatosis to “non-alcoholic steatohepatitis (NASH)”, an advanced form of NAFLD characterized by sterile inflammation and progressive fibrosis. Despite being the fastest growing cause for liver transplantation, no effective anti-NASH treatments are available today.

Metabolic interactions amongst the lipid-damaged hepatocytes, Kupffer cells (KCs), and hepatic stellate cells (HSCs) underlies NASH initiation. Identifying NASH-promoting metabolite crosstalk can lead to novel therapeutic targets. However, studying metabolite crosstalk in livers remains challenging because separating different liver cell types is time-consuming and severely disturbs their metabolism.

To overcome metabolic disturbances, we developed a “NASH-in-a-dish” system, an innovative system that models NASH initiation in vitro for metabolite crosstalk interrogation. NASH-in-a-dish is a co-culture of mouse hepatocyte organoids, primary KCs, and primary HSCs, in which steatohepatitis can be induced and effects of specific metabolites can be tested. Uniquely, NASH-in-a-dish enables rapid separation of constituent cell types (<2 minutes) for metabolite profiling.