Human Enteroid Monolayers: A Novel, Functionally-Stable Model for Investigating Oral Drug Disposition
To further the development of an in vitro model that can more faithfully recapitulate drug disposition of orally administered drugs, we investigated the utility of human enteroid monolayers to simultaneously assess intestinal drug absorption and first-pass metabolism processes. We cultured human enteroid monolayers from three donors, derived via biopsies containing duodenal stem cells that were propagated and then differentiated atop permeable Transwell® inserts, and confirmed transformation into a largely enterocyte population via RNA-seq analysis and immunocytochemical (ICC) assays. Proper cell morphology was assessed and confirmed via bright field microscopy and ICC imaging of tight junction proteins and other apically and basolaterally localized proteins. Enteroid monolayer barrier integrity was demonstrated by elevated transepithelial electrical resistance (TEER) that stabilized after 10 days in culture and persisted for 42 days. These results were corroborated by low paracellular transport probe permeability at 7 and 21 days in culture. The activity of a prominent drug metabolizing enzyme, CYP3A, was confirmed at 7, 21, and 42 days culture under basal, 1?,25(OH)2 vitamin D3-induced, and 6',7'-dihydroxybergamottin-inhibited conditions. The duration of these experiments is particularly noteworthy, as this is the first study assessing DMET function for enteroids cultured for greater than 12 days. The sum of these results suggests enteroid monolayers are a promising in vitro model to investigate and quantitatively predict an orally administered drug's intestinal absorption and/or metabolism and associated drug-drug/natural product-drug interactions. Overall design: Matched enteroid monolayer and undifferentiated spheroids from three donors were compared to characterize differences and infer suitability of enteroid monolayers as a model for oral drug disposition.