Establishment of a Hepatocyte-Kupffer Cell Coculture Model for Assessment of Proinflammatory Cytokine Effects on Metabolizing Enzymes and Drug Transporters

Abstract

Elevated levels of proinflammatory cytokines associated with infection and inflammation can modulate cytochrome P450 enzymes, leading to potential disease-drug interactions and altered small-molecule drug disposition. We established a human-derived hepatocyte-Kupffer cell (Hep:KC) coculture model to assess the indirect cytokine impact on hepatocytes through stimulation of KC-mediated cytokine release and compared this model with hepatocytes alone. Characterization of Hep:KC cocultures showed an inflammation response after treatment with lipopolysaccharide and interleukin (IL)-6 (indicated by secretion of various cytokines). Additionally, IL-6 exposure upregulated acutephase proteins (C-reactive protein, alpha-1-acid glycoprotein, and serum amyloid A2) and downregulated CYP3A4. Compared with hepatocytes alone, Hep:KC cocultures showed enhanced IL-1b-mediated effects but less impact from both IL-2 and IL-23. Hep:KC cocultures treated with IL-1b exhibited a higher release of proinflammatory cytokines, an increased upregulation of acute-phase proteins, and a larger extent of metabolic enzyme and transporter suppression. IC 50 values for IL-1b-mediated CYP3A4 suppression were lower in Hep:KC cocultures (98.0-144 pg/ml) compared with hepatocytes alone (IC 50 > 5000 pg/ml). Cytochrome suppression was preventable by blocking IL-1b interaction with IL-1R1 using an antagonist cytokine or an anti-IL-1b antibody. Unlike IL-1b, IL-6-mediated effects were comparable between hepatocyte monocultures and Hep:KC cocultures. IL-2 and IL-23 caused a negligible inflammation response and a minimal inhibition of CYP3A4. In both hepatocyte monocultures and Hep:KC cocultures, IL-2RB and IL-23R were undetectable, whereas IL-6R and IL-1R1 levels were higher in Hep:KC cocultures. In summary, compared with hepatocyte monocultures, the Hep:KC coculture system is a more robust in vitro model for studying the impact of proinflammatory cytokines on metabolic enzymes.

IL-36–Induced Toxicity in Neonatal Mice Involves TNF-α Production by Liver Myeloid Cells

“…In contrast with these findings and in agreement with published data, Il1r1 mRNA was expressed at similar levels in isolated hepatocytes as in total liver (Fig. 3E) (29,30). Furthermore, total liver Il1r1 mRNA levels remained constant at different ages (Fig.…”

Prediction of Drug Clearance and Drug-Drug Interactions in Microscale Cultures of Human Hepatocytes

“…Incorporation of liver stromal cells in MPCCs may allow prediction of drug clearance rates under disease states such as fibrosis and inflammation (Nguyen et al, 2015). Furthermore, miniaturization into a 384-well format should enable higher throughput screening in MPCCs.…”

Modeling Therapeutic Antibody–Small Molecule Drug-Drug Interactions Using a Three-Dimensional Perfusable Human Liver Coculture Platform

“…KCs were included to build a more complete hepatic cellular microenvironment, and numerous studies have shown their impact on the behavior of in vitro liver systems, such as enhanced IL-1b-mediated cytokine secretion, acute phase protein production, and cytochrome P450 suppression (Hoebe et al, 2001;Sunman et al, 2004;Nguyen et al, 2015;Sarkar et al, 2015;Rose et al, 2016). There are other mechanisms mediated by IL-6 that could potentially be modeled in the LiverChip system such as drug transporter activity.…”

“…This receptor exists in both a cell membrane-bound form and a shed soluble form (sIL-6R), both of which bind IL-6 with equivalent affinity and mediate the same downstream signaling cascade (Rose-John, 2012;Scheller et al, 2014). C-reactive protein (CRP), an acute phase protein secreted by hepatocytes in response to IL-6, is commonly used as an in vitro marker of IL-6 signaling in hepatocytes (Bode et al, 2012;Nguyen et al, 2015).…”