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Publication Detail

Title: Detection of phospholipidosis induction: a cell-based assay in high-throughput and high-content format.

Authors: Shahane, Sampada A; Huang, Ruili; Gerhold, David; Baxa, Ulrich; Austin, Christopher P; Xia, Menghang

Published In J Biomol Screen, (2014 Jan)

Abstract: Drug-induced phospholipidosis is characterized by the accumulation of intracellular phospholipids in cells exposed to cationic amphiphilic drugs. The appearance of unicentric or multicentric multilamellar bodies viewed under an electron microscope (EM) is the morphological hallmark of phospholipidosis. Although the EM method is the gold standard for detecting cellular phospholipidosis, this method has its drawbacks, including low throughput, high cost, and unsuitability for screening a large chemical library. In this study, a cell-based phospholipidosis assay has been developed using the LipidTOX Red reagent in HepG2 cells and miniaturized into a 1536-well plate format. To validate this assay for high-throughput screening (HTS), the LOPAC library of 1280 compounds was screened using a quantitative HTS platform. A group of known phospholipidosis inducers, such as amiodarone, propranolol, chlorpromazine, desipramine, promazine, clomipramine, and amitriptyline, was identified by the screen, consistent with previous reports. Several novel phospholipidosis inducers, including NAN-190, ebastine, GR127935, and cis-(Z)-flupentixol, were identified in this study and confirmed using the EM method. These results demonstrate that this assay can be used to evaluate and profile large numbers of chemicals for drug-induced phospholipidosis.

PubMed ID: 24003057 Exiting the NIEHS site

MeSH Terms: Hep G2 Cells; High-Throughput Screening Assays*; Humans; Intracellular Space/metabolism; Lipidoses/chemically induced; Lipidoses/diagnosis*; Lipidoses/metabolism*; Microscopy, Electron, Transmission; Phospholipids/metabolism*; Reproducibility of Results

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