Abstract
The utilisation of genome-wide transcriptomics has played a pivotal role in advancing the field of toxicology, allowing the mapping of transcriptional signatures to chemical exposures. These activities have uncovered several transcriptionally regulated pathways that can be utilised for assessing the perturbation impact of a chemical and also the identification of toxic mode of action. However, current transcriptomic platforms are not very amenable to high-throughput workflows due to, high cost, complexities in sample preparation and relatively complex bioinformatic analysis. Thus, transcriptomic investigations are usually limited in dose and time dimensions and are, therefore, not optimal for implementation in risk assessment workflows. In this study, we investigated a new cost-effective, transcriptomic assay, TempO-Seq, which alleviates the aforementioned limitations. This technique was evaluated in a 6-compound screen, utilising differentiated kidney (RPTEC/TERT1) and liver (HepaRG) cells and compared to non-transcriptomic label-free sensitive endpoints of chemical-induced disturbances, namely phase contrast morphology, xCELLigence and glycolysis. Non-proliferating cell monolayers were exposed to six sub-lethal concentrations of each compound for 24 h. The results show that utilising a 2839 gene panel, it is possible to discriminate basal tissue-specific signatures, generate dose–response relationships and to discriminate compound-specific and cell type-specific responses. This study also reiterates previous findings that chemical-induced transcriptomic alterations occur prior to cytotoxicity and that transcriptomics provides in depth mechanistic information of the effects of chemicals on cellular transcriptional responses. TempO-Seq is a robust transcriptomic platform that is well suited for in vitro toxicity experiments.
Original language | English |
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Pages (from-to) | 2517-2531 |
Number of pages | 15 |
Journal | Archives of Toxicology |
Volume | 92 |
Issue number | 8 |
Early online date | 14 Jul 2018 |
DOIs | |
Publication status | Published - Aug 2018 |
Funding
Acknowledgements The study was funded by the 7th Framework project DETECTIVE (Grant no. 266838 to PJ, BvdW, MV), the Horizon 2020 project EU-ToxRisk (http://www.eu-toxrisk.eu/ Grant no. 681002, to BvdW, PJ), the European Research Council (ERC Starting Grant 335476 to MV), the Center for Alternatives to Animal Testing at Johns Hopkins University Baltimore-USA (to MV), the Tiroler Wissen-schaftsfonds (Grant no. UNI-0404/1768, to AW) and the Long Range Initiative Innovative Science Award of the European Chemical Industry Council (CEFIC, 2015 to AL). We would like to thank Pranika Singh for helping with data processing. The study was funded by the 7th Framework project DETECTIVE (Grant no. 266838 to PJ, BvdW, MV), the Horizon 2020 project EU-ToxRisk (http://www.eu-toxrisk.eu/ Grant no. 681002, to BvdW, PJ), the European Research Council (ERC Starting Grant 335476 to MV), the Center for Alternatives to Animal Testing at Johns Hopkins University Baltimore-USA (to MV), the Tiroler Wissenschaftsfonds (Grant no. UNI-0404/1768, to AW) and the Long Range Initiative Innovative Science Award of the European Chemical Industry Council (CEFIC, 2015 to AL). We would like to thank Pranika Singh for helping with data processing. The co-authors P. Shepard, B. Seligmann, H. C. VanSteenhouse and J. M. Yeakley are owners/employees of BioSpyder Technologies Inc., the manufacturer of the TempO-Seq assay. All biological experiments and data interpretation was done independently of BioSpyder. None of the other co-authors are associated with this company financially or otherwise.
Funders | Funder number |
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7th Framework project | |
BioSpyder Technologies Inc. | |
Tiroler Wissen-schaftsfonds | |
Center for Alternatives to Animal Testing, Johns Hopkins Bloomberg School of Public Health | |
European Chemical Industry Council | |
Seventh Framework Programme | 335476, 681002, 266838 |
European Research Council | |
Horizon 2020 | |
Landes Tirols | UNI-0404/1768 |
Keywords
- Dedifferentiation
- HepaRG
- RPTEC/TERT1
- Stress responses
- TempO-Seq