Development of a neurotoxicity assay that is tuned to detect mitochondrial toxicants

Research output: Contribution to journalJournal article – Annual report year: 2019Researchpeer-review

Documents

DOI

  • Author: Delp, Johannes

    University of Konstanz, Germany

  • Author: Funke, Melina

    University of Konstanz, Germany

  • Author: Rudolf, Franziska

    University of Konstanz, Germany

  • Author: Cediel, Andrea

    Karolinska Institutet, Sweden

  • Author: Bennekou, Susanne Hougaard

    Group for Chemical Risk Assessment and GMO, National Food Institute, Technical University of Denmark, Kemitorvet, 2800, Kgs. Lyngby, Denmark

  • Author: van der Stel, Wanda

    Leiden University, Netherlands

  • Author: Carta, Giada

    Vrije Universiteit Amsterdam, Netherlands

  • Author: Jennings, Paul

    Vrije Universiteit Amsterdam, Netherlands

  • Author: Toma, Cosimo

    IRCCS Istituto di ricerche farmacologiche Mario Negri - Milano, Bergamo, Ranica, Italy

  • Author: Gardner, Iain

    Certara, United Kingdom

  • Author: van de Water, Bob

    Leiden University, Netherlands

  • Author: Forsby, Anna

    Karolinska Institutet, Sweden

  • Author: Leist, Marcel

    University of Konstanz, Germany

View graph of relations

Many neurotoxicants affect energy metabolism in man, but currently available test methods may still fail to predict mito- and neurotoxicity. We addressed this issue using LUHMES cells, i.e., human neuronal precursors that easily differentiate into mature neurons. Within the NeuriTox assay, they have been used to screen for neurotoxicants. Our new approach is based on culturing the cells in either glucose or galactose (Glc-Gal-NeuriTox) as the main carbohydrate source during toxicity testing. Using this Glc-Gal-NeuriTox assay, 52 mitochondrial and non-mitochondrial toxicants were tested. The panel of chemicals comprised 11 inhibitors of mitochondrial respiratory chain complex I (cI), 4 inhibitors of cII, 8 of cIII, and 2 of cIV; 8 toxicants were included as they are assumed to be mitochondrial uncouplers. In galactose, cells became more dependent on mitochondrial function, which made them 2-3 orders of magnitude more sensitive to various mitotoxicants. Moreover, galactose enhanced the specific neurotoxicity (destruction of neurites) compared to a general cytotoxicity (plasma membrane lysis) of the toxicants. The Glc-Gal-NeuriTox assay worked particularly well for inhibitors of cI and cIII, while the toxicity of uncouplers and non-mitochondrial toxicants did not differ significantly upon glucose ↔ galactose exchange. As a secondary assay, we developed a method to quantify the inhibition of all mitochondrial respiratory chain functions/complexes in LUHMES cells. The combination of the Glc-Gal-NeuriTox neurotoxicity screening assay with the mechanistic follow up of target site identification allowed both, a more sensitive detection of neurotoxicants and a sharper definition of the mode of action of mitochondrial toxicants.
Original languageEnglish
JournalArchives of Toxicology
Volume93
Issue number6
Pages (from-to)1585-1608
ISSN0340-5761
DOIs
Publication statusPublished - 2019
CitationsWeb of Science® Times Cited: No match on DOI

    Research areas

  • High content imaging, High-throughput toxicity screening, Mechanistic safety assessment, Metabolic reprogramming, Mitotoxicity, Neurotoxicity

Download statistics

No data available

ID: 183320296