Compound cytotoxicity profiling using quantitative high-throughput screening - PubMed (original) (raw)

. 2008 Mar;116(3):284-91.

doi: 10.1289/ehp.10727.

Ruili Huang, Kristine L Witt, Noel Southall, Jennifer Fostel, Ming-Hsuang Cho, Ajit Jadhav, Cynthia S Smith, James Inglese, Christopher J Portier, Raymond R Tice, Christopher P Austin

Affiliations

Compound cytotoxicity profiling using quantitative high-throughput screening

Menghang Xia et al. Environ Health Perspect. 2008 Mar.

Abstract

Background: The propensity of compounds to produce adverse health effects in humans is generally evaluated using animal-based test methods. Such methods can be relatively expensive, low-throughput, and associated with pain suffered by the treated animals. In addition, differences in species biology may confound extrapolation to human health effects.

Objective: The National Toxicology Program and the National Institutes of Health Chemical Genomics Center are collaborating to identify a battery of cell-based screens to prioritize compounds for further toxicologic evaluation.

Methods: A collection of 1,408 compounds previously tested in one or more traditional toxicologic assays were profiled for cytotoxicity using quantitative high-throughput screening (qHTS) in 13 human and rodent cell types derived from six common targets of xenobiotic toxicity (liver, blood, kidney, nerve, lung, skin). Selected cytotoxicants were further tested to define response kinetics.

Results: qHTS of these compounds produced robust and reproducible results, which allowed cross-compound, cross-cell type, and cross-species comparisons. Some compounds were cytotoxic to all cell types at similar concentrations, whereas others exhibited species- or cell type-specific cytotoxicity. Closely related cell types and analogous cell types in human and rodent frequently showed different patterns of cytotoxicity. Some compounds inducing similar levels of cytotoxicity showed distinct time dependence in kinetic studies, consistent with known mechanisms of toxicity.

Conclusions: The generation of high-quality cytotoxicity data on this large library of known compounds using qHTS demonstrates the potential of this methodology to profile a much broader array of assays and compounds, which, in aggregate, may be valuable for prioritizing compounds for further toxicologic evaluation, identifying compounds with particular mechanisms of action, and potentially predicting in vivo biological response.

Keywords: 1,536-well; NTP 1,408 compound library; PubChem; RT-CES; cell viability; qHTS.

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Figures

Figure 1

Figure 1

Intraexperiment reproducibility. The figure shows example replicate dose–response curves for colchicine, cycloheximide, progesterone, tetraethylene glycol diacrylate, and sodium dichromate dehydrate in rat primary kidney proximal tubule cells.

Figure 2

Figure 2

Pharmacological profile of compound activity. (A) Percentage of activity in each class identified from all compounds in 13 cell lines. (B) Potency distribution of all compounds in 13 cell lines.

Figure 3

Figure 3

Compound activity patterns clustered by cell and species type. The compounds with an IC50 of < 92 μM in at least one cell type are selected and arranged in the order of a hierarchical clustering based on their IC50 values as shown in the dendrogram on the left side of the heat map. Neighboring compounds share similar activity patterns. In the figure, each row represents a compound, and each column is a cell type. Compound activity in each cell line is colored according to potency (IC50) range. Potent compounds are deeper shades of red and nontoxic compounds are white. Assays are also clustered by similarity in their compound IC50 patterns as shown in the dendrogram on the top of the heat map.

Figure 4

Figure 4

Compound activity across different species. Compound activity in each cell line is colored according to potency (IC50) range. The figure shows examples of compounds that are more cytotoxic to human cells (top two rows) or to rodent cells (middle rows), and compounds with similar levels of cytotoxicity in human and rodent cells (bottom rows).

Figure 5

Figure 5

Kinetics of cytotoxicity responses for digitonin (A), potassium dichromate (B), cycloheximide (C), doxorubicin (D), and tamoxifen (E) in HepG2 cells monitored by the RT-CES system. 10,000 cells/well were plated in 16-well strips for the RT-CES cytotoxicity assay.↓, time of compound addition. Different compound concentrations are indicated by different colors. Data are normalized to the time of compound addition at 22–24 hr of cell culture.

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