Integrated Isogenic Human Induced Pluripotent Stem Cell-Based Liver and Heart Microphysiological Systems Predict Unsafe Drug-Drug Interaction.

Felipe T Lee-Montiel,Hideaki Okochi,Steven C Boggess,Verena Charwat,Holger Willenbring,Nathaniel Huebsch,Evan W Miller,Alexander Laemmle,Kevin E Healy,Laure Dumont,Brian Siemons,Caleb S Lee,Matthew J Hancock,Ishan Goswami

doi:10.3389/fphar.2021.667010

Abstract

Three-dimensional (3D) microphysiological systems (MPSs) mimicking human organ function in vitro are an emerging alternative to conventional monolayer cell culture and animal models for drug development. Human induced pluripotent stem cells (hiPSCs) have the potential to capture the diversity of human genetics and provide an unlimited supply of cells. Combining hiPSCs with microfluidics technology in MPSs offers new perspectives for drug development. Here, the integration of a newly developed liver MPS with a cardiac MPS—both created with the same hiPSC line—to study drug–drug interaction (DDI) is reported. As a prominent example of clinically relevant DDI, the interaction of the arrhythmogenic gastroprokinetic cisapride with the fungicide ketoconazole was investigated. As seen in patients, metabolic conversion of cisapride to non-arrhythmogenic norcisapride in the liver MPS by the cytochrome P450 enzyme CYP3A4 was inhibited by ketoconazole, leading to arrhythmia in the cardiac MPS. These results establish integration of hiPSC-based liver and cardiac MPSs to facilitate screening for DDI, and thus drug efficacy and toxicity, isogenic in the same genetic background.

Highlights

The drug development pipeline generally starts with a broad collection of candidate compounds that are narrowed down by traditional in vitro drug screening tools including biochemical analysis and cell-based assays, for example, using immortalized cell lines, computational modeling, and testing in animal models
Immunofluorescence showed lack of the pluripotency marker OCT3/4 past the Human induced pluripotent stem cells (hiPSCs) stage (Figure 1B); hepatic nuclear factor 4 alpha (HNF4A) started to be expressed at day 13 when cells committed to hepatic fate; alpha-fetoprotein (AFP) stained strongly positive in immature hepatocytes at day 18; and albumin (ALB) was only detectable at the final stage of differentiation
For the uptake transporter studies, hiPSC-Heps were exposed to a specific drug internalized by the transporter, with or without a specific inhibitor of the transporter—activity of organic anion transporters (OATs) was tested with acyclovir/probenecid, organic cation transporters (OCTs) with metformin/ decynium-22, and organic anion transporting polypeptides (OATPs) with rosuvastatin/rifamycin-SV

Summary

Introduction

The drug development pipeline generally starts with a broad collection of candidate compounds that are narrowed down by traditional in vitro drug screening tools including biochemical analysis and cell-based assays, for example, using immortalized cell lines, computational modeling, and testing in animal models. Too often compounds pass the go/no-go risk decisions based on animal model data, but fail later in clinical trials due to species-specific differences in physiology. The worstcase scenario occurs when drugs make it through the full screening pipeline and clinical trials to market, only to be recalled due to unforeseen side effects in some patients, a prominent example being the arrhythmogenic effect of the gastroprokinetic cisapride (Okada et al, 2015). In addition to the inability of animal models to accurately mimic human physiology, such failures are due to lack of diversity in the human genetic backgrounds tested in single cell lines, and lack of diversity in phase I and II clinical trials

Methods

Results

Conclusion