Comparison of base-line and chemical-induced transcriptomic responses in HepaRG and RPTEC/TERT1 cells using TempO-Seq

Area of Science:

• Toxicology and Molecular Biology
• In vitro models and high-throughput screening

Background:

• Genome-wide transcriptomics aids toxicology by mapping transcriptional signatures to chemical exposures.
• Current transcriptomic platforms face limitations in high-throughput workflows due to cost, sample preparation, and bioinformatics complexity.
• These limitations restrict dose and time dimensions, hindering optimal use in risk assessment.

Purpose of the Study:

• To investigate TempO-Seq, a novel, cost-effective transcriptomic assay designed to overcome limitations of existing platforms.
• To evaluate TempO-Seq's efficacy in a 6-compound screen using differentiated kidney and liver cells.
• To compare TempO-Seq with non-transcriptomic, label-free endpoints for detecting chemical-induced disturbances.

Main Methods:

• Utilized differentiated kidney (RPTEC/TERT1) and liver (HepaRG) cells exposed to six sub-lethal compound concentrations for 24 hours.
• Employed a 2839-gene panel with the TempO-Seq assay.
• Compared TempO-Seq results with phase contrast morphology, xCELLigence, and glycolysis measurements.

Main Results:

• TempO-Seq successfully discriminated basal tissue-specific signatures and compound-specific responses.
• Dose-response relationships were established for chemical exposures.
• Transcriptomic alterations were observed to precede cytotoxicity, providing mechanistic insights into chemical effects.

Conclusions:

• TempO-Seq is a robust and cost-effective transcriptomic platform suitable for high-throughput in vitro toxicity screening.
• The assay provides valuable mechanistic information on cellular transcriptional responses to chemical exposures.
• TempO-Seq enhances the application of transcriptomics in risk assessment workflows.