A hollow fiber membrane-based liver organoid-on-a-chip model for examining drug metabolism and transport

Myszczyszyn, A.; Muench, A.; Lehmann, V.; Sinnige, T.; van Steenbeek, F. G.; Bouwmeester, M.; Samsom, R.-A.; Keuper-Navis, M.; van der Made, T. K.; Kogan, D.; Braem, S.; van der Laan, L. J. W.; Eslami Amirabadi, H.; van de Steeg, E.; Masereeuw, R.; Spee, B.

doi:10.1101/2024.08.12.607504

bioRxiv · 10.1101/2024.08.12.607504

A hollow fiber membrane-based liver organoid-on-a-chip model for examining drug metabolism and transport

Myszczyszyn, A.·Muench, A.·Lehmann, V.·Sinnige, T.·van Steenbeek, F. G.·Bouwmeester, M.·Samsom, R.-A.·Keuper-Navis, M.·van der Made, T. K.·Kogan, D.·Braem, S.·van der Laan, L. J. W.·Eslami Amirabadi, H.·van de Steeg, E.·Masereeuw, R.·Spee, B.

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Abstract

Liver-on-a-chip models predictive for both metabolism as well as canalicular and blood transport of drug candidates in humans are lacking. Here, we established an advanced, bioengineered and animal component-free hepatocyte-like millifluidic system based on 3D hollow fiber membranes (HFMs), recombinant human laminin 332 coating and adult human stem cell-derived organoids. Organoid fragments formed polarized and tight monolayers on HFMs with improved hepatocyte-like maturation, as compared to standard 3D organoid cultures in Matrigel from matched donors. Gene expression profiling and immunofluorescence revealed that hepatocyte-like monolayers expressed a broad panel of phase I (e.g., CYP3A4, CYP2D6) and II (UGTs, SULTs) drug-metabolizing enzymes and drug transporters (e.g., OATP1B3, MDR1 and MRP3). Moreover, statically cultured monolayers displayed phase I and II metabolism of a cocktail of six relevant compounds, including midazolam and 7-hydroxycoumarin. We also demonstrated the disposition of midazolam in the basal/blood-like circulation and apical/canalicular-like compartment of the millifluidic chip. Finally, we connected the system to the other two PK/ADME-most relevant organ systems, i.e. small intestine- and kidney proximal tubule-like to study the bioavailability of midazolam and coumarin, and excretion of metformin. In conclusion, we generated a proof-of-concept liver organoid-on-a-chip model for examining metabolism and transport of drugs, which can be further developed to predict PK/ADME profiles in humans.

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Myszczyszyn, A., Muench, A., Lehmann, V., Sinnige, T., van Steenbeek, F. G., Bouwmeester, M., Samsom, R.-A., Keuper-Navis, M., van der Made, T. K., Kogan, D., Braem, S., van der Laan, L. J. W., Eslami Amirabadi, H., van de Steeg, E., Masereeuw, R., Spee, B.. 2024-08-15. A hollow fiber membrane-based liver organoid-on-a-chip model for examining drug metabolism and transport. https://doi.org/10.1101/2024.08.12.607504

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