3D human liver spheroids for translational pharmacology and toxicology

Abstract

Drug development is a failure‐prone endeavour, and more than 85% of drugs fail during clinical development, showcasing that current preclinical systems for compound selection are clearly inadequate. Liver toxicity remains a major reason for safety failures. Furthermore, all efforts to develop pharmacological therapies for a variety of chronic liver diseases, such as non‐alcoholic steatohepatitis (NASH) and fibrosis, remain unsuccessful. Considering the time and expense of clinical trials, as well as the substantial burden on patients, new strategies are thus of paramount importance to increase clinical success rates. To this end, human liver spheroids are becoming increasingly utilized as they allow to preserve patient‐specific phenotypes and functions for multiple weeks in culture. We here review the recent application of such systems for i) predictive and mechanistic analyses of drug hepatotoxicity, ii) the evaluation of hepatic disposition and metabolite formation of low clearance drugs and iii) the development of drugs for metabolic and infectious liver diseases, including NASH, fibrosis, malaria and viral hepatitis. We envision that with increasing dissemination, liver spheroids might become the new gold standard for such applications in translational pharmacology and toxicology.

Evaluation of a Three-Dimensional Primary Human Hepatocyte Spheroid Model: Adoption and Industrialization for the Enhanced Detection of Drug-Induced Liver Injury

“…Physicochemical properties of drugs do vary, and if there are concerns regarding compound deposition, then ToF-SIMS could be used to assess this. In conclusion, it is likely a combination of the maintenance of these phenotypic features, along with the extended compound exposure duration, which appears to be responsible for the improved potential for DILI prediction, and such features may also lead to the increased use of spheroids as in vitro models for liver diseases …”

“…These biological advantages, together with the proven capture of increased number of severely hepatotoxic compounds, made it evident that this was the model of choice, with scope for even further enhancement. While at present, the readout is a valuable but simple ATP measurement, the addition of biochemical and mechanistic end points (such as mitochondrial dysfunction or oxidative stress) could further enhance this assay and could aid in the investigation of mechanisms of toxicity. ,, The introduction of nonparenchymal cells to include an immune component has also been suggested for the potential to improve prediction of DILI. ,, Additionally, the incorporation of a dynamic perfusion system across spheroids, rather than bathing hepatocytes in a static environment for long durations, could potentially better represent the pharmacokinetics of the in vivo situation.…”

An Engineered Protein-Based Building Block (Albumin Methacryloyl) for Fabrication of a 3D In Vitro Cryogel Model

“…The limitations of 2D culture systems have driven the development of 3D in vitro culture systems that are categorized into nonscaffold-based systems such as spheroid/cell aggregate culture [ 14 , 15 ] and hanging drop [ 16 , 17 ] and scaffold-based systems such as microfluidic liver models [ 18 ], co-culture [ 19 ], and 3D printing [ 11 , 20 ] to mimic the complex features of the liver such as enhanced cell–cell communications, multicellular environments, and efficient supply of nutrients and oxygen. Especially, spheroid culture systems can provide excellent cell–cell interactions in the multicellular structure [ 21 ], but the formation of necrosis in the center of large spheroids needs to be overcome. The hanging drop method is an efficient tool for generating cell spheroids/aggregates.…”

Current Research Trends in the Application of In Vitro Three-Dimensional Models of Liver Cells

“…The spheroid formation of HepaRG and HepG2 cells can be improved through incubation in the liver biomatrix scaffolds produced in decellularized natural mice. The HepaRG cells used in the experiment showed the further enhancement of the spheroids, but a recent study using 150 compounds concluded that HepG2 spheroids are more sensitive to hepatotoxic drugs [ 126 ].…”