High-throughput compatible rat liver microtissues toassess idiosyncratic toxicological effectsIntroduction         Althoug...
High-throughput compatible rat liver microtissues toassess idiosyncratic toxicological effectsInflammation-mediated toxici...
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White Paper Idiosyncratic Toxicology

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White Paper Idiosyncratic Toxicology

  1. 1. High-throughput compatible rat liver microtissues toassess idiosyncratic toxicological effectsIntroduction Although idiosyncratic adverse drug reactions are rare they represent a significant risk forhealthcare and industry. Liver inflammatory cells, in addition to genetic and environmentalfactors, are known to mediate such adverse reactions. So far, inflammation-mediated toxicityhas only been successfully reproduced experimentally in animal models. InSphero’s organotypicrat liver microtissue (rLi MT) platform provides the first high throughput in vitro model shown tobe capable of detecting inflammation-related idiosyncratic effects. The toxicological effect ofTrovafloxacin and Ranitidine were tested either with or without Kupffer cell stimulation usingLPS. An increased toxicological effect could clearly be seen in the LPS-treated cultures. Predictingidiosyncratic effects is a serious challenge for the pharmaceutical industry. Here, for the firsttime, we show that a robust and affordable in vitro liver model, designed for high throughputdata generation, is capable of reproducing inflammation-mediated idiosyncratic toxicity.Functional characterization of rLi microtissuesScaffold-free liver microtissues were produced from primary hepatocytes and non-parenchymalcells (NPC) comprising Kupffer macrophages and endothelial cells. Prior to MT formation inhanging drops, hepatocytes and NPCs were mixed in MT re-aggregation medium. Fig.1 Mitochondrial activity of rat liver microtissues was stable for 5 weeks in culture, as shown by stable ATP content over time, in contrast to the corresponding 2D-sandwich cultures from the same source (A). To assess metabolic functionality CYP3A activity was induced by the addition of 10 µM Dexamethasone for 48 hours. CYP3A- inducibility maturated over time and was stable for 28 days in culture (B). Albumin secretion in rLi MTs was 4-5x higher than in sandwich cultures (C). To test functionality of Kupffer- macrophages, the MTs were stimulated by Lipopolysaccharides (LPS). Elevated levels of Interleukin-6 (IL-6) were detected only in the co-culture MTs (D).InSphero AG, Zurich, Switzerland, www.insphero.com, sales@insphero.com, Phone +41-44-515049-0 1
  2. 2. High-throughput compatible rat liver microtissues toassess idiosyncratic toxicological effectsInflammation-mediated toxicityInflammation has been shown to decrease the threshold for hepatoxicity of certain drugs. Herewe tested 2 drugs ,Trovafloxin and Ranitidine, both known to induce inflammation-mediatedtoxicity. The presence of inflammatory cells in the liver microtissue model allows native-likeinflammation conditions to be reproduced, which, in turn, makes it possible to simulateidiosyncratic drug toxicity.Fig.2 Mitochondrial activity (A, C) andcytotoxicity (B, D) of rLi MTs treated withTrovafloxacin and Ranitidine. Both drugswere incubated with and without LPS for48h. Both end points reflect that theaddition of LPS impacts the toxicologicaleffects of both drugs as compared to theDMSO control. .Idiosyncratic adverse drug reactions affect various tissues, but the liver is one of the prime targetorgans. Of the 28 drugs withdrawn from the U.S. market between 1976 and 2005, 6 werewithdrawn due to hepatoxicity. Therefore, model systems to detect adverse drug reactionsduring drug development is a prime focus for the pharmaceutical industry. However, currentpreclinical testing protocols often fail to identify drugs that cause idiosyncratic adverse drugreactions, due to inappropriate model systems. Currently, only animal models have the ability toreflect inflammation-mediated idiosyncratic toxicity. The rLi MT model has demonstrated itsability to predict inflammation-induced hepatoxicity. This model, designed for high throughputdata generation, is easy to implement in drug testing campaigns for efficient early stage drugde-risking.InSphero AG, Zurich, Switzerland, www.insphero.com, sales@insphero.com, Phone +41-44-515049-0 2

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