Modeling Therapeutic Antibody–Small Molecule Drug-Drug Interactions Using a Three-Dimensional Perfusable Human Liver Coculture Platform

Abstract

Traditional in vitro human liver cell culture models lose key hepatic functions such as metabolic activity during short-term culture. Advanced three-dimensional (3D) liver coculture platforms offer the potential for extended hepatocyte functionality and allow for the study of more complex biologic interactions, which can improve and refine human drug safety evaluations. Here, we use a perfusion flow 3D microreactor platform for the coculture of cryopreserved primary human hepatocytes and Kupffer cells to study the regulation of cytochrome P450 3A4 isoform (CYP3A4) activity by chronic interleukin 6 (IL-6)-mediated inflammation over 2 weeks. Hepatocyte cultures remained stable over 2 weeks, with consistent albumin production and basal IL-6 levels. Direct IL-6 stimulation that mimics an inflammatory state induced a dose-dependent suppression of CYP3A4 activity, an increase in C-reactive protein (CRP) secretion, and a decrease in shed soluble interleukin-6 receptor (IL-6R) levels, indicating expected hepatic IL-6 bioactivity. Tocilizumab, an anti-IL-6R monoclonal antibody used to treat rheumatoid arthritis, has been demonstrated clinically to impact small molecule drug pharmacokinetics by modulating cytochrome P450 enzyme activities, an effect not observed in traditional hepatic cultures. We have now recapitulated the clinical observation in a 3D bioreactor system. Tocilizumab was shown to desuppress CYP3A4 activity while reducing the CRP concentration after 72 hours in the continued presence of IL-6. This change in CYP3A4 activity decreased the half-life and area under the curve up to the last measurable concentration (AUC last ) of the small molecule CYP3A4 substrate simvastatin hydroxy acid, measured before and after tocilizumab treatment. We conclude that next-generation in vitro liver culture platforms are well suited for these types of long-term treatment studies and show promise for improved drug safety assessment.

Integrated gut/liver microphysiological systems elucidates inflammatory inter‐tissue crosstalk

“…While most OOC systems are fabricated from polydimethylsiloxane (PDMS)-a versatile elastomer that is easy to prototype, has good optical properties, and is oxygen permeablehydrophobic compounds, including steroid hormones and many drugs, strongly partition into PDMS, thus precluding quantitative analysis and control of drug exposures (Toepke and Beebe, 2006). We therefore use a platform fabricated from polysulfone via micromachining, with on-board pneumatic microfluidic pumping (Domansky et al, 2010;Inman et al, 2007) adapted from technology now used commercially in the Liverchip TM for extended 3D culture of functional liver tissue (Long et al, 2016;Sarkar et al, 2015Sarkar et al, , 2017Tsamandouras et al, 2017;Vivares et al, 2015), and for demonstration of human liver CYP450 regulation by inflammation (Long et al, 2016). This on-board pumping technology minimizes space, auxiliary equipment, and dead volumes associated with excess tubing (Domansky et al, 2010;Inman et al, 2007).…”

“…We focus here on gut‐liver interactions, as gut‐liver crosstalk is an integral part of normal physiology and its dysregulation is a common denominator in many disease conditions (Marshall, ). Gut and liver are major organs involved in drug absorption and metabolism, and changes to their functional interactions, such as those precipitated by injury or disease, can impact their responses to therapeutic intervention (Deng et al, ; Long et al, ; Morgan, ). The liver receives most of its blood supply from the gut via the portal circulation so it is constantly exposed to gut‐derived factors, including metabolites, microbial antigens, and inflammatory mediators.…”

Quantitative Assessment of Population Variability in Hepatic Drug Metabolism Using a Perfused Three-Dimensional Human Liver Microphysiological System

“…The LiverChip comprises a scaffold that fosters formation of an array of ∼0.2-mm 3D tissue structures from primary human liver cells, and an on-board microfluidic pumping system, driven by pneumatics, that precisely perfuses the scaffold with culture medium to control oxygenation and shear stress on the tissue, enabling long-term culture with retention of physiological responses (Dash et al, 2009; Domansky et al, 2010; Vivares et al, 2015). The LiverChip platform, seeded with hepatocytes or with mixtures of hepatocytes and nonparenchymal cells, has been applied to analyze drug metabolism, inflammatory effects, drug-drug interactions, and as a model of breast cancer metastasis to the liver (Wheeler et al, 2014; Sarkar et al, 2015; Vivares et al, 2015; Long et al, 2016). …”

3D Miniaturization of Human Organs for Drug Discovery

“…A very interesting approach to model the complex sinusoid-like structures using 3D perfused hepatic co-cultures in a microfluidic system has been developed by Griffith and co-workers. [74][75][76][77] This microfluidic model includes key features such as adjustable flow rates based on oxygen consumption and long-term steady maintenance of the oxygen gradient. This comprehensive system, using co-cultures of diverse heterogeneous cells, forms liver …”

“…[53] In these multicellular aggregates, the need for supporting gels or matrices is eliminated, the adverse effects 3D culture approach for generating a laminated cerebral cortex like structure from pluripotent stem cells. [57,58] Microfabrication Neuroprogenitor cells Microfluidic culture platform containing a relief pattern of soma and axonal compartments connected by microgrooves to direct, isolate, lesion, and biochemically analyze CNS axons [67,68] 3D bioprinting Primary human cortical neurons Discrete layers of primary neutrons in a RGD peptide-modified gellan gum [118][119][120] Intestine (Gut) Self-assembled Stem cells Identified intestinal stem cells and differentiated cells in vitro [59,60] Microfabrication Human epithelial cells Mimic contractility by using mechanochemical actuator [11,19,27,72] Liver Self-assembled Human stem cells 3D culture of self-renewing human liver tissue [61,62] Microfabrication Hepatocytes and fibroblasts Microengineered hepatic microtissues containing hepatocytes and fibroblasts [73][74][75][76][77] 3D bioprinting HepG2 and HUVEC Multilayered organ tissue model [96,[155][156][157] Vessel Microfabrication Rat brain endothelial cells 3D culture in microfluidic device [63][64][65][66] 3D bioprinting HUVECs and HUVSMCs Scaffold-less vessel formation using spheroid fusion [84][85][86][87][88][89][90][91]…”