Development of an in vitro human liver system for interrogating nonalcoholic steatohepatitis
Ryan E. Feaver, … , Brian R. Wamhoff, Ajit Dash
Ryan E. Feaver, … , Brian R. Wamhoff, Ajit Dash
Published December 8, 2016
Citation Information: JCI Insight. 2016;1(20):e90954. https://doi.org/10.1172/jci.insight.90954.
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Technical Advance Hepatology Therapeutics

Development of an in vitro human liver system for interrogating nonalcoholic steatohepatitis

Abstract

A barrier to drug development for nonalcoholic steatohepatitis (NASH) is the absence of translational preclinical human-relevant systems. An in vitro liver model was engineered to incorporate hepatic sinusoidal flow, transport, and lipotoxic stress risk factors (glucose, insulin, free fatty acids) with cocultured primary human hepatocytes, hepatic stellate cells (HSCs), and macrophages. Transcriptomic, lipidomic, and functional endpoints were evaluated and compared with clinical data from NASH patient biopsies. The lipotoxic milieu promoted hepatocyte lipid accumulation (4-fold increase, P < 0.01) and a lipidomics signature similar to NASH biopsies. Hepatocyte glucose output increased with decreased insulin sensitivity. These changes were accompanied by increased inflammatory analyte secretion (e.g., IL-6, IL-8, alanine aminotransferase). Fibrogenic activation markers increased with lipotoxic conditions, including secreted TGF-β (>5-fold increase, P < 0.05), extracellular matrix gene expression, and HSC activation. Significant pathway correlation existed between this in vitro model and human biopsies. Consistent with clinical trial data, 0.5 μM obeticholic acid in this model promoted a healthy lipidomic signature, reduced inflammatory and fibrotic secreted factors, but also increased ApoB secretion, suggesting a potential adverse effect on lipoprotein metabolism. Lipotoxic stress activates similar biological signatures observed in NASH patients in this system, which may be relevant for interrogating novel therapeutic approaches to treat NASH.

Authors

Ryan E. Feaver, Banumathi K. Cole, Mark J. Lawson, Stephen A. Hoang, Svetlana Marukian, Brett R. Blackman, Robert A. Figler, Arun J. Sanyal, Brian R. Wamhoff, Ajit Dash

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Figure 7

Obeticholic acid restores homeostasis in the human liver lipotoxic system.

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Obeticholic acid restores homeostasis in the human liver lipotoxic syste...
Nonparenchymal cells (NPCs) and hepatocytes on device were exposed to the lipotoxic milieu containing 0.5 μM obeticholic acid (OCA) or DMSO vehicle control (Veh) for 10 days. (A) Secreted FGF19 was measured in the media effluent from devices at day 10. n = 3 experiments, 1 donor. (B) Nile red staining intensity from hepatocyte images were quantified and represented as fold change relative to healthy controls. n = 4 experiments, 1 donor. (C) Lipids from hepatocytes from device exposed to the healthy or lipotoxic milieu with 0.5 μM OCA or vehicle control were measured by metabolomics. Scatterplot representation of differentially expressed lipids in these hepatocytes are shown as log2-fold change and colored by response similarity index (RSI). Triacyglycerols (○), cholesterol esters (×), all other lipids (•). n = 4 experiments, 2 donors. (D and E) Secreted analytes were measured in the media effluent from devices at day 10. n ≥ 5 experiments, 3 donors. (F) Secreted apolipoproteins were measured in the media effluent from devices at day 10. n = 4 experiments, 2 donors. Triangles indicate samples that were below the lower limit of quantification. *P < 0.05, **P < 0.01, Student’s 2-tailed t test.