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

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Quantitative Assessment of Population Variability in Hepatic Drug Metabolism Using a Perfused Three-Dimensional Human Liver Microphysiological System S Quantitative Assessment of Population Variability in Hepatic Drug Metabolism Using a Perfused Three- Dimensional Human Liver Microphysiological System The MIT Faculty has made this article openly available. Please share how this access benefits you. Your story matters. Citation Tsamandouras, N., T. Kostrzewski, C. L. Stokes, L. G. Griffith, D. J. Hughes, and M. Cirit. “Quantitative Assessment of Population Variability in Hepatic Drug Metabolism Using a Perfused Three- Dimensional Human Liver Microphysiological System.” Journal of Pharmacology and Experimental Therapeutics 360, no. 1 (October 19, 2016): 95–105. As Published http://dx.doi.org/10.1124/JPET.116.237495 Publisher American Society for Pharmacology & Experimental Therapeutics (ASPET) Version Final published version Citable link http://hdl.handle.net/1721.1/117617 Terms of Use Creative Commons Attribution-NonCommercial 4.0 International Detailed Terms http://creativecommons.org/licenses/by-nc/4.0/ Downloaded from jpet.aspetjournals.org at ASPET Journals on August 30, 2018 95 105, January 2017 – and the associ- ” http://dx.doi.org/10.1124/jpet.116.237495 c clearance, as assessed in vitro, g metabolism in the context of a J Pharmacol Exp Ther 360:95 average individual, “ Several in vitro systems have been traditionally applied to ated interindividual variability is not obtained. variability in intrinsic metaboli needstobecoupledwiththepopulationvariabilityassociated with other physiologic processes indrug vivo binding (e.g., to hepatic plasma bloodpredictions proteins, flow, etc.). at Thus, to theapproach perform population (Trame in vivo et level,results al., a are 2016) integrated systems is into pharmacology desirable,pharmacokinetic population physiologically (PBPK) where based models the (Rostami-Hodjegan, inJones and 2012; vitro Rowland-Yeo, 2013; Tsamandouras et al., 2015a,b). study drug metabolism,(Obach, including 1999) human and liver cryopreservedsions human microsomes (Brown hepatocyte et al., suspen- 2007).very Although valuable these systems in have drugeasy been development to use, (Di they etstudy lose al., metabolic 2012) of activity and over low-clearanceObach, time, are 2015; hence compounds Hutzler the et isin al., challenging 2015). vitro In (Di addition,(Hallifax systems overall et and these al., tend 2010).often Finally, to drug-metabolism performed studies underpredict in are pooled human in liver from microsomes vivo severalclearance or clearance donors. hepatocytes refers Thus, to an the predicted intrinsic variability in intrinsic metabolic clearancephenacetin ranged to from 66.8% 24.1% for for propranolol (expressedvariation). as coefficient Albumin, of urea,levels LDH, were identified and asbolic cytochrome clearance. significant Predicted P450 predictors clearance values of mRNA physiological from in the system liver vitro were micro- meta- correlatedvalues. with A the population observed inmodel physiologically was vivo based developed for pharmacokinetic lidocainethe to illustrate in the vitro translation of in output to vivo. the Stochasticpredicted observed the pharmacokinetic simulations observed variability clinical withthe concentration-time associated population this profiles variability. and This modelpopulation is variability the successfully first in study dru microphysiological of system andthe has use of important these systems implications during for the drug development process. s but also accounting ” Supplemental material to this article can be found at: HERAPEUTICS T in vivo translation. For each ny other types of variability med independently for hepa- – http://jpet.aspetjournals.org/content/suppl/2016/10/19/jpet.116.237495.DC1 T. Kostrzewski, C. L. Stokes, L. G. Griffith, D. J. Hughes, and M. Cirit XPERIMENTAL E 1 Introduction average individual “ CV, coefficient of variation; 3D, three-dimensional; IDV, interdonor variability; IWV, interwell variability; LDH, lactate 105$25.00 – HARMACOLOGY AND P 2016 by The Author(s) ª This article has supplemental material available at jpet.aspetjournals.org. OURNAL OF J N.T. and T.K. contributed equally to this work. s To investigate interindividual variability in vitro, drug- During preclinical drug development, prediction of hepatic dx.doi.org/10.1124/jpet.116.237495. 1 This work was supported by the DARPA Microphysiological Systems HE for the associated population variability (Jamei et al., 2009). metabolism assays must betocytes perfor obtained from differentstatistical donors analysis and with of an thedonor appropriate obtained variability in data intrinsic tosurement clearance estimate error/uncertainty disentangled the and from inter- mea- (e.g., a interwell). Additionally, the estimate of interindividual This is an open access article distributed under the CC BY-NC Attribution 4.0 International license. Copyright clearance is of significantdose importance and to guide the set selectiondrug the concentrations of first within dosage the human therapeutic regimens window. thatto However, achieve efficiently designhepatic clinical drug studies, metabolism it andonly at is human the level pharmacokinetics, crucial of an not to predict Program [Grant W911NF-12-2-0039]Microphysiological and Systems Program the [Grant National 4-UH3-TR000496-03]. Institutes of Health ABBREVIATIONS: dehydrogenase; MPS, microphysiologicalchain system; reaction; P450, PT, parallel cytochrome tube; P450; WS, well PBPK, stirred. physiologically based pharmacokinetic; PCR, polymerase T Received August 30, 2016; accepted October 17, 2016 ABSTRACT In this work, we first describedrug the population metabolism variability in using hepatic different donors cryopreserved cultured in a hepatocytes perfusedliver three-dimensional from human microphysiological five system,sulting data and can then beframework show integrated to with how accomplish a in the modeling vitro re- and simulation N. Tsamandouras, Department of Biological Engineering,CN Massachusetts Bio Institute Innovations, of Hertfordshire, Technology, United Cambridge, Kingdom Massachusetts (T.K., (N.T., L.G.G., D.J.H.); M.C.); and Stokes Consulting, Redwood City, California (C.L.S.) Microphysiological System donor, metabolic depletion profilesetin, of diclofenac, six lidocaine, compounds ibuprofen, (phenac- olone) propranolol, were and measured, prednis- alonglevels with of 90 metabolite metabolism-related formation, genes, and mRNA viability markers [lactate of dehydrogenase functional (LDH) release, albumin,production]. and urea Drug depletioneffects modeling. Substantial data interdonor variability were was observed with analyzed respect to withother gene mixed- expression measured levels, hepatocyte drug functions. metabolism, Specifically, and interdonor Metabolism Using a Perfused Three-Dimensional Human Liver Quantitative Assessment of Population Variability in Hepatic Drug 1521-0103/360/1/95 96 Tsamandouras et al. Recently, new hepatic in vitro culture models have emerged sacrificed at day 6 for RNA analysis. After the initial attachment to improve physiologic responses and mitigate the rapid loss of period, cells undergo morphogenesis to form an array of 3D micro- metabolic function typically observed in culture, thus offering tissues within the channels of the scaffold over a period of 3 days. ’ opportunity to improve predictions of human drug clearance, Cells were maintained in Williams E medium containing primary especially for low-clearance compounds. Whereas static two- hepatocyte thawing and plating supplements (Life Technologies) for the first day of culture. Maintenance supplements (Life Technolo- dimensional cocultures of hepatocytes with other cell types gies), which are serum-free, were used thereafter. All cultures were show some stabilization of function, a variety of three- maintained in a standard humidified atmosphere at 37°C with 5% dimensional (3D) culture models incorporating perfusion flow CO2 and had a first complete medium change at 24 hours, then after a exhibit prolonged viability and function under serum-free condi- further 72 hours. tions (Ebrahimkhani et al., 2014). Such 3D perfused models of Hepatocyte Culture Phenotypic Characterization. Albumin liver and other tissues, where microfluidic or microscale reactors and urea production as well as lactate dehydrogenase (LDH) release areusedtocontroltheflowofculturemedium,areoftentermed were measured before the drug-metabolism study (4 days postseed- “microphysiological systems” (MPS) or “organs on chips.” In this ing). Albumin production was measured in supernatant using a human work, we use a particular well developed, commercially available albumin enzyme-linked immunosorbent assay (Assay Pro, St. Charles, microreactor system for 3D perfused liver culture, the LiverChip MO). Urea was quantified with a colorimetric assay kit (BioAssay Systems, Hayward, CA) and LDH secretion was measured using the (CN Bio Innovations, Hertfordshire, UK) (Dash et al., 2009; CytoTox 96 nonradioactive cytotoxicity assay (Promega, Southampton, Domansky et al., 2010; Sarkar et al., 2015; Vivares et al., 2015). UK). Albumin, urea, and LDH were also measured postdose at the end The LiverChip comprises a scaffold that fosters formation of an of the drug-metabolism study (day 5 for wells treated with phenacetin; Downloaded from array of ∼0.2-mm 3D tissue structures from primary human liver day 6 for wells treated with diclofenac, propranolol, lidocaine, and cells, and an on-board microfluidic pumping system, driven by ibuprofen; and day 7 for wells treated with prednisolone). At the end
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