Method Profile
Measure parent compounds and selected metabolites across a human engineered intestinal barrier–liver cell-line chip, including CYP3A4 inhibitor challenges.
Investigate coupled intestinal drug transfer, metabolic activity and specific drug–drug or compound-level drug–food interactions.
Genome-edited human Caco-2 cells in the upper channel and CYPs-UGT1A1 knock-in human HepG2 cells in the lower channel; wild-type matched cell-line chip as comparator.
Two-channel PDMS device; 14-day model formation, with rocking at 0.1 cycle/min applied only during absorption/metabolism assays; upper/lower sampling at 2, 4, 24 and 48 hours.
Engineered and wild-type cell pairs
Caco-2 barrier and HepG2 aggregates; Lucifer yellow permeability and marker characterization; no primary tissue or donor-diverse cohort.
Drug and material controls
Six parent drugs and selected CYP metabolites measured by LC–MS/MS; separate PDMS/polystyrene sorption checks. Itraconazole or bergamottin at 10 μM before exposure to 10 μM triazolam.
Compare genome-edited versus wild-type chips and static versus rocking conditions. Interpret parent disappearance with measured metabolite formation and separate material losses; reported error bars are SD.
Six-drug absorption/metabolism comparison plus CYP3A4 inhibitor challenge.
Characterization n = 3 devices; drug profiles n = 4 devices; inhibitor challenge n = 3 devices.
Wild-type chip, static conditions, no-inhibitor control and material-only sorption controls.
Edited chips produced higher selected CYP metabolites. Itraconazole and bergamottin raised triazolam and lowered α-hydroxytriazolam at 24 and 48 hours in both channels. Lower-channel diclofenac did not significantly differ between edited and wild-type chips despite metabolite differences.
Engineered human Caco-2 and HepG2 cells supported simultaneous measurement of parent drugs and metabolites in a two-channel chip. Itraconazole and bergamottin increased triazolam concentrations while reducing its CYP3A4 metabolite, with material sorption and assay timing limiting pharmacokinetic interpretation.
