Abstract
Non-alcoholic steatohepatitis (NASH) is a progressive and severe liver disease, characterized by lipid accumulation, inflammation, and downstream fibrosis. Despite its increasing prevalence, there is no approved treatment yet available for patients. This has been at least partially due to the lack of predictive preclinical models for studying this complex disease. Here, we present a 3D in vitro microtissue model that uses spheroidal, scaffold free co-culture of primary human hepatocytes, Kupffer cells, liver endothelial cells and hepatic stellate cells. Upon exposure to defined and clinically relevant lipotoxic and inflammatory stimuli, these microtissues develop key pathophysiological features of NASH within 10 days, including an increase of intracellular triglyceride content and lipids, and release of pro-inflammatory cytokines. Furthermore, fibrosis was evident through release of procollagen type I, and increased deposition of extracellular collagen fibers. Whole transcriptome analysis revealed changes in the regulation of pathways associated with NASH, such as lipid metabolism, inflammation and collagen processing. Importantly, treatment with anti-NASH drug candidates (Selonsertib and Firsocostat) decreased the measured specific disease parameter, in accordance with clinical observations. These drug treatments also significantly changed the gene expression patterns of the microtissues, thus providing mechanisms of action and revealing therapeutic potential. In summary, this human NASH model represents a promising drug discovery tool for understanding the underlying complex mechanisms in NASH, evaluating efficacy of anti-NASH drug candidates and identifying new approaches for therapeutic interventions.
Highlights
Non-alcoholic steatohepatitis (NASH) is a progressive and severe liver disease, characterized by lipid accumulation, inflammation, and downstream fibrosis
Non-alcoholic fatty liver disease (NAFLD) defines a spectrum of pathologies, ranging from simple steatosis to its most severe form: non-alcoholic steatohepatitis (NASH), where it is believed that lipid accumulation in the liver and gut derived endotoxins results in inflammatory and fibrotic p rocesses[1,2,3]
We demonstrate that the prolonged co-culture of primary human hepatocytes (PHHs), Kupffer cells (KCs), liver endothelial cells (LECs) and hepatic stellate cells (HSCs), exposed to Free fatty acids (FFA) and LPS, allows longitudinal changes that approximate disease progression and mimic the observed features of NASH
Summary
Non-alcoholic steatohepatitis (NASH) is a progressive and severe liver disease, characterized by lipid accumulation, inflammation, and downstream fibrosis. P rill[21] developed a 3D model that enables the long-term culture of primary human hepatocytes (PHHs) and induction of lipid loading upon exposure to defined stimuli While these 2D and 3D monoculture approaches allow a recapitulation of the early stages of NAFLD i.e. lipid metabolism, they cannot be used to investigate compound effects on downstream events in inflammation and fibrosis, since they lack all the relevant cells involved[14]. To address this, another elegant approach was developed which incorporated PHHs, KCs, and HSCs. To address this, another elegant approach was developed which incorporated PHHs, KCs, and HSCs In this model, the PHHs were cultured in a transwell format under flow conditions, whereby inflammatory and fibrotic pathways were activated using lipotoxic s timuli[22]. Since the cells were cultured in a 2D format, with PHHs physically separated from NPCs, this model lacked adequate crosstalk between cells
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
