Capturing time-dependent activation of genes and stress-response pathways using transcriptomics in iPSC-derived renal proximal tubule cells

Paul Jennings, Paul Jennings, Pranika Singh, Daniel da Costa Pereira, Anita Feher, Andras Dinnyes, Thomas E Exner, Anja Wilmes

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

Transcriptomic analysis is a powerful method in the utilization of New Approach Methods (NAMs) for identifying mechanisms of toxicity and application to hazard characterization. With this regard, mapping toxicological events to time of exposure would be helpful to characterize early events. Here, we investigated time-dependent changes in gene expression levels in iPSC-derived renal proximal tubular-like cells (PTL) treated with five diverse compounds using TempO-Seq transcriptomics with the aims to evaluate the application of PTL for toxicity prediction and to report on temporal effects for the activation of cellular stress response pathways. PTL were treated with either 50 μM amiodarone, 10 μM sodium arsenate, 5 nM rotenone, or 300 nM tunicamycin over a temporal time course between 1 and 24 h. The TGFβ-type I receptor kinase inhibitor GW788388 (1 μM) was used as a negative control. Pathway analysis revealed the induction of key stress-response pathways, including Nrf2 oxidative stress response, unfolding protein response, and metal stress response. Early response genes per pathway were identified much earlier than 24 h and included HMOX1, ATF3, DDIT3, and several MT1 isotypes. GW788388 did not induce any genes within the stress response pathways above, but showed deregulation of genes involved in TGFβ inhibition, including downregulation of CYP24A1 and SERPINE1 and upregulation of WT1. This study highlights the application of iPSC-derived renal cells for prediction of cellular toxicity and sheds new light on the temporal and early effects of key genes that are involved in cellular stress response pathways.

Original languageEnglish
Pages (from-to)1773-1793
Number of pages21
JournalCell biology and toxicology
Volume39
Issue number4
Early online date31 Dec 2022
DOIs
Publication statusPublished - Aug 2023

Bibliographical note

Funding Information:
This research was funded by the EU project in3 a Marie Sklodowska-Curie Action-Innovative Training Network under grant no. 721975. Additional funding was received from the EU H2020 research and innovation programme under grant agreement no. 739593 (HCEMM for AD).

Publisher Copyright:
© 2022, The Author(s).

Funding

This research was funded by the EU project in3 a Marie Sklodowska-Curie Action-Innovative Training Network under grant no. 721975. Additional funding was received from the EU H2020 research and innovation programme under grant agreement no. 739593 (HCEMM for AD).

FundersFunder number
EU H2020 Research and Innovation Programme
Horizon 2020 Framework Programme739593
Horizon 2020 Framework Programme
H2020 Marie Skłodowska-Curie Actions
European Commission721975
European Commission

    Keywords

    • iPSC
    • NAMs
    • Renal proximal tubular
    • Stress-response pathways
    • Temporal gene expression
    • Transcriptomics

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