Temporal transcriptomic alterations of cadmium exposed human iPSC-derived renal proximal tubule-like cells

Pranika Singh, Vidya Chandrasekaran, Barry Hardy, Anja Wilmes, Paul Jennings*, Thomas E. Exner

*Corresponding author for this work

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

Cadmium is a well-studied environmental pollutant where the kidney and particularly the proximal tubule cells are especially sensitive as they are exposed to higher concentrations of cadmium than other tissues. Here we investigated the temporal transcriptomic alterations (TempO-Seq) of human induced pluripotent stem cell (iPSC)-derived renal proximal tubule-like (PTL) cells exposed to 5 μM cadmium chloride for 1, 2, 4, 8, 12, 16, 20, 24, 72 and 168 h. There was an early activation (within 4 h) of the metal and oxidative stress responses (metal-responsive transcription factor-1 (MTF1) and nuclear factor erythroid-2-related factor 2 (Nrf2) genes). The Nrf2 response returned to baseline within 24 h. The Activator Protein 1 (AP-1) regulated genes HSPA6 and FOSL-1 followed the Nrf2 time course. While the MTF1 genes also spiked at 4 h, they remained strongly elevated over the entire exposure period. The data and cell culture model utilised will be useful in further research aimed at the refinement of safe human exposure limits for cadmium, other metals and their mixtures.

Original languageEnglish
Article number105229
Pages (from-to)1-13
Number of pages13
JournalToxicology in Vitro
Volume76
Early online date3 Aug 2021
DOIs
Publication statusPublished - Oct 2021

Bibliographical note

Funding Information:
This work was funded by the EU project in3 a Marie Sklodowska-Curie Action - Innovative Training Network under grant no. 721975 .

Funding Information:
The human iPSC line SBAD2 clone 1, generated using Cytotune 2.0 (Thermofisher) from dermal fibroblasts within the IMI funded StemBANCC project (grant agreement no 115439, http://stembancc.org ), ( Morrison et al., 2015 ) was utilised here. These cells were further genetically manipulated by Prof. A. Dinnyés laboratory, Biotalentum, Hungary to contain a Green Fluorescent Protein (GFP) tag downstream of one allele of the HMOX-1 gene and designated as SBAD2 HMOX1-eGFP (although, this fluorescent property was not utilised in this study). SBAD2 HMOX1-eGFP iPSCs were cultured on Geltrex coated plates (Life Technologies A1413302), in mTeSR1 medium (StemCell Technologies 05850) and were routinely passaged with EDTA (0.02% Versene, Lonza BE17-711E) as previously described ( Wilmes et al., 2017 ).

Publisher Copyright:
© 2021 The Authors

Funding

This work was funded by the EU project in3 a Marie Sklodowska-Curie Action - Innovative Training Network under grant no. 721975 . The human iPSC line SBAD2 clone 1, generated using Cytotune 2.0 (Thermofisher) from dermal fibroblasts within the IMI funded StemBANCC project (grant agreement no 115439, http://stembancc.org ), ( Morrison et al., 2015 ) was utilised here. These cells were further genetically manipulated by Prof. A. Dinnyés laboratory, Biotalentum, Hungary to contain a Green Fluorescent Protein (GFP) tag downstream of one allele of the HMOX-1 gene and designated as SBAD2 HMOX1-eGFP (although, this fluorescent property was not utilised in this study). SBAD2 HMOX1-eGFP iPSCs were cultured on Geltrex coated plates (Life Technologies A1413302), in mTeSR1 medium (StemCell Technologies 05850) and were routinely passaged with EDTA (0.02% Versene, Lonza BE17-711E) as previously described ( Wilmes et al., 2017 ).

FundersFunder number
Horizon 2020 Framework Programme721975
European Commission
Innovative Medicines Initiative115439

    Keywords

    • Cadmium
    • HSPA6
    • In vitro
    • iPSC
    • Metal responsive transcription factor-1
    • Metallothionein
    • Nuclear factor erythroid-2-related factor 2
    • Proximal tubules
    • Temporal expression patterns

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