Establishment of an In Vitro High-Throughput Screening Assay for Detecting Phospholipidosis-Inducing Potential

Kasahara, Toshihiko; Tomita, Kazuo; Murano, Hiroyuki; Harada, Tsuyoshi; Tsubakimoto, Keisuke; Ogihara, Takuo; Ohnishi, Syuhei; Kakinuma, Chihaya

doi:10.1093/toxsci/kfj067

Cited by 76 publications

(69 citation statements)

References 11 publications

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“…The same cutoff val- ue (1.5-fold) was also reported by Bauch et al (2015) using HepG2 and H9c2 cells. In addition, the relationship between concentrations of PLD-positive compounds and fluorescence intensity of phosphatidylcholine-conjugated dye taken up by CHO-K1 cells was investigated by Kasahara et al (2006). Kasahara et al (2006) reported that the minimum positive concentrations for the PLD assay were 10 μM (13.8 μM in our assay) in amiodarone, 10 μM (11 μM in our assay) in chlorpromazine, 10 μM (5.9 μM in our assay) in imipramine, 3 μM (6.8 μM in our assay) in tamoxifen.…”

Section: Discussionmentioning

confidence: 99%

“…In vitro models for predicting drug-induced PLD have been developed using various cells such as HepG2 cells (Kasahara et al, 2006;Tilmant et al, 2011;van de Water et al, 2011), human primary hepatocytes (Gum et al, 2001;Kasahara et al, 2006), and rat primary hepatocytes (Gum et al, 2001;Kasahara et al, 2006;Takagi et al, 2016). However, the HepG2 cell line is devoid of a substantial and variable set of liver-specific functions, particularly the major cytochrome P450s (CYPs) that are involved in xenobiotic metabolism (Guguen-Guillouzo and Guillouzo, 2010).…”

Section: Introductionmentioning

confidence: 99%

“…The study authors demonstrated that metabolic activation is the key factor for PLD induction in loratadine. In addition, loratadine did not induce PLD in Chinese hamster ovary (CHO-K1) cells (Kasahara et al, 2006). Thus, a cell-model maintaining drug metabolism function may be important for predicting drug-induced PLD in loratadine.…”

Section: Introductionmentioning

confidence: 99%

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A cell-based assay using HepaRG cells for predicting drug-induced phospholipidosis

2017

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-The utility of HepaRG cells as an in vitro cell-based assay system for predicting druginduced phospholipidosis (PLD) was investigated. In experiment 1, 10 PLD-positive compounds and 11 PLD-negative compounds were selected. HepaRG cells were treated with each compound for 48 hr. In experiment 2, loratadine and desloratadine, a major metabolite of loratadine, were used to assess metabolic activation for PLD. HepaRG cells were treated with loratadine and desloratadine in the presence or absence of 500 μM 1-aminobenzotriazole (ABT), a broad CYP inhibitor, for 48 hr. After treatment with compounds in experiments 1 and 2, the relative fluorescence intensity (RFI) was measured using LYSO-ID Red dye to assess the PLD induction. In experiment 1, our cell-based assay system using HepaRG cells exhibited 100% sensitivity and 100% specificity for predicting drug-induced PLD. In experiment 2, loratadine increased the RFI in the PLD assay. However, the increase in the RFI was not observed in co-treatment with loratadine and ABT. In addition, desloratadine increased the RFI in the presence and absence of ABT. These results suggested that metabolic activation of loratadine may contribute to PLD in HepaRG cells. We newly demonstrated that HepaRG cells have a high ability for predicting drug-induced PLD. In addition, we newly showed that HepaRG cells may predict drug-induced PLD mediated by metabolic activation of loratadine. Thus, a cell-based assay system using HepaRG cells is a useful model for predicting drug-induced PLD.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A cell-based assay using HepaRG cells for predicting drug-induced phospholipidosis

2017

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“…An independent test found that based on these genes it is possible to obtain reliable predictions for the PPL potential of drugs [52]. High-throughput screening with cell-based assays represents an effective tool to evaluate large sets of compounds in the preclinical stages to determine their PPL potential [53]. A high-throughput physicochemical assay for PPL screening in early phases of drug discovery is based on the measurement of drug-phospholipid complex formation observed by their effect on the critical micelle concentration of a short-chain acidic phospholipid [54].…”

Section: Drugs With Phospholipidosis Inducing Potentialmentioning

confidence: 99%

Weka Machine Learning for Predicting the Phospholipidosis Inducing Potential

Ivanciuc

2008

CTMC

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Abstract:The drug discovery and development process is lengthy and expensive, and bringing a drug to market may take up to 18 years and may cost up to 2 billion $US. The extensive use of computer-assisted drug design techniques may considerably increase the chances of finding valuable drug candidates, thus decreasing the drug discovery time and costs. The most important computational approach is represented by structure-activity relationships that can discriminate between sets of chemicals that are active/inactive towards a certain biological receptor. An adverse effect of some cationic amphiphilic drugs is phospholipidosis that manifests as an intracellular accumulation of phospholipids and formation of concentric lamellar bodies. Here we present structure-activity relationships (SAR) computed with a wide variety of machine learning algorithms trained to identify drugs that have phospholipidosis inducing potential. All SAR models are developed with the machine learning software Weka, and include both classical algorithms, such as k-nearest neighbors and decision trees, as well as recently introduced methods, such as support vector machines and artificial immune systems. The best predictions are obtained with support vector machines, followed by a perceptron artificial neural network, logistic regression, and k-nearest neighbors.

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“…Although the relationship between DIPL and adverse effects in humans remains unknown, it is necessary to consider toxicity triggered by DIPL in humans. Some studies regarding in vitro screening to avoid DIPL have been reported (Sawada et al, 2005;Kasahara et al, 2006;Morelli et al, 2006;Zhou et al, 2011;Takagi et al, 2016). However, to evaluate DIPL in experimental animals, it is important to consider species differences, organ selectivity, and affinity for phospholipids.…”

Section: Introductionmentioning

confidence: 99%

Assessment of amiodarone-induced phospholipidosis in chimeric mice with a humanized liver

et al. 2017

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-It is important to consider susceptibility to drug-induced toxicity between animals and humans. Chimeric mice with a humanized liver are expected to predict hepatotoxicity in humans. Druginduced phospholipidosis (DIPL), in which phospholipids accumulate, is a known entity. In this study, we examined whether chimeric mice can reveal species differences in DIPL. Changes in various phosphatidylcholine (PhC) molecules were investigated in the liver of chimeric mice after administering amiodarone, which induces phospholipidosis. Liquid chromatography-tandem mass spectrometry revealed that levels of PhCs tended to increase in the liver after administration of amiodarone. The liver of chimeric mice consists of human hepatocytes and residual mouse hepatocytes. We used imaging mass spectrometry (IMS) to evaluate the increase of PhCs in human and mouse hepatocytes after administration of amiodarone. IMS visualizes localization of endogenous and exogenous molecules in tissues. The IMS analysis suggested that the localized levels of several PhCs tended to be higher in the human hepatocytes than those in mouse hepatocytes, and PhC levels changed in response to amiodarone. Chimeric mice with a humanized liver will be useful to evaluate species differences in DIPL between mice and humans.

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Establishment of an In Vitro High-Throughput Screening Assay for Detecting Phospholipidosis-Inducing Potential

Cited by 76 publications

References 11 publications

A cell-based assay using HepaRG cells for predicting drug-induced phospholipidosis

A cell-based assay using HepaRG cells for predicting drug-induced phospholipidosis

Weka Machine Learning for Predicting the Phospholipidosis Inducing Potential

Assessment of amiodarone-induced phospholipidosis in chimeric mice with a humanized liver

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