Gut-Liver Microphysiological Systems Revealed Arachidonic Acid as a Potential Crosstalk Factor Modulating Drug Metabolism

Coculture of hepatocytes with small intestine cells using MPS have been shown to enhance hepatic drug metabolism, yet the crosstalk mechanism is still unclear. Coculture of PXB-cells and iPS-derived intestine cells in MPS with perfusion and direct oxygenation were conducted to investigate this crosstalk. Analysis on albumin secretion, CYP enzyme actvity and differentially expressed genes confirmed that perfusion and direct oxygenation enhanced PXB-cells. Upregulation of "epoxygenase P450 pathway" GO terms indicated involvement of Arachidonic Acid (ARA) in the crosstalk mechanism. This is further confirmed by comparing the effect of ARA and coculture toward PXB-cells CYP enzymes activity. We hypothesize that some stimulus from PXB-cells resulted made iPS-derived intestine cells released ARA in form of lipoprotein, which is then taken in by PXB-cells and enhanced CYP activity. Overall design: PXB-cells were cultured in normal 24-well tissue culture plate (TCPS), MPS with perfusion only (MPS O2[-]) and MPS with perfusion + direct oxygenation (MPS O2[+]). To observe crosstalk, RNA-seq was performed by comparing cocultured and monocultured PXB-cells