Prediction of Drug-Induced Liver Injury in Micropatterned Co-cultures Containing iPSC-Derived Human Hepatocytes

Abstract

Primary human hepatocytes (PHHs) are a limited resource for drug screening, their quality for in vitro use can vary considerably across different lots, and a lack of available donor diversity restricts our understanding of how human genetics affect drug-induced liver injury (DILI). Induced pluripotent stem cell-derived human hepatocyte-like cells (iPSC-HHs) could provide a complementary tool to PHHs for high-throughput drug screening, and ultimately enable personalized medicine. Here, we hypothesized that previously developed iPSC-HH-based micropatterned co-cultures (iMPCCs) with murine embryonic fibroblasts could be amenable to long-term drug toxicity assessment. iMPCCs, created in industry-standard 96-well plates, were treated for 6 days with a set of 47 drugs, and multiple functional endpoints (albumin, urea, ATP) were evaluated in dosed cultures against vehicle-only controls to enable binary toxicity decisions. We found that iMPCCs correctly classified 24 of 37 hepatotoxic drugs (65% sensitivity), while all 10 non-toxic drugs tested were classified as such in iMPCCs (100% specificity). On the other hand, conventional confluent cultures of iPSC-HHs failed to detect several liver toxins that were picked up in iMPCCs. Results for DILI detection in iMPCCs were remarkably similar to published data in PHH-MPCCs (65% versus 70% sensitivity) that were dosed with the same drugs. Furthermore, iMPCCs detected the relative hepatotoxicity of structural drug analogs and recapitulated known mechanisms of acetaminophen toxicity in vitro. In conclusion, iMPCCs could provide a robust tool to screen for DILI potential of large compound libraries in early stages of drug development using an abundant supply of commercially available iPSC-HHs.

Requirements for human iPS cell-derived hepatocytes as an alternative to primary human hepatocytes for assessing absorption, distribution, metabolism, excretion and toxicity of pharmaceuticals

“…Further studies are needed to find the best combination of these culture methods and cell types and improve the functions of hiPSC-derived hepatocytes. Recent studies reported that the micro-patterned co-culture methods enhanced the hepatic functions of hiPSC-derived hepatocytes and could be useful to screen for DILI (Berger et al, 2014;Ware et al, 2015). They seeded hiPSC-derived hepatocytes on the micro-patterned collagen in a microwell plate, and co-cultured them with murine fibroblasts.…”

Idiosyncratic Drug-Induced Liver Injury (IDILI): Potential Mechanisms and Predictive Assays

“…While iPSCs require dedifferentiation with subsequent redifferentiation to derive appropriate cell lines, they provide an innovative way to both model liver disease and assess how certain polymorphisms may affect drug metabolism and disposition in vitro. Additionally, human iPSCs (hiPSCs) have a significant amount of commercial availability for applications in liver toxicity testing and have the potential to be used in coculture with hepatocytes and fibroblasts to give an accurate model for predicting potential liver toxicity [199–201]. …”

Translational Prospects and Challenges in Human Induced Pluripotent Stem Cell Research in Drug Discovery

“…More importantly, one of the most desirable characteristics needed for improving translational prospects of these cells is sustained functionality of a handful of the most important hepatic cytochrome P450 enzymes, since these CYP proteins are involved in metabolism and bioactivation of more than 75% of drugs [94]. Despite the fact that human PSC-derived hepatocytes are still characterized by lower CYP enzymatic activity when compared to fresh primary hepatocytes, these cells have already been demonstrated to be a valuable platform for hepatocyte-specific toxicity assays [95,96,97,98] (Table 1). Moreover, iPSC-derived hepatocytes exhibit high levels of other hepatic functions, such as synthesis and secretion of various plasma proteins (carrier proteins like albumin, blood clotting factors and apolipoproteins), glycogen storage, urea production, and bile production.…”