Abstract
| Original language | English |
|---|---|
| Journal | Archives of Toxicology |
| Volume | 93 |
| Issue number | 6 |
| Pages (from-to) | 1585-1608 |
| ISSN | 0340-5761 |
| DOIs | |
| Publication status | Published - 2019 |
Keywords
- High content imaging
- High-throughput toxicity screening
- Mechanistic safety assessment
- Metabolic reprogramming
- Mitotoxicity
- Neurotoxicity
Cite this
}
Development of a neurotoxicity assay that is tuned to detect mitochondrial toxicants. / Delp, Johannes; Funke, Melina; Rudolf, Franziska; Cediel, Andrea; Bennekou, Susanne Hougaard; van der Stel, Wanda; Carta, Giada; Jennings, Paul; Toma, Cosimo; Gardner, Iain; van de Water, Bob; Forsby, Anna; Leist, Marcel.
In: Archives of Toxicology, Vol. 93, No. 6, 2019, p. 1585-1608.Research output: Contribution to journal › Journal article › Research › peer-review
TY - JOUR
T1 - Development of a neurotoxicity assay that is tuned to detect mitochondrial toxicants
AU - Delp, Johannes
AU - Funke, Melina
AU - Rudolf, Franziska
AU - Cediel, Andrea
AU - Bennekou, Susanne Hougaard
AU - van der Stel, Wanda
AU - Carta, Giada
AU - Jennings, Paul
AU - Toma, Cosimo
AU - Gardner, Iain
AU - van de Water, Bob
AU - Forsby, Anna
AU - Leist, Marcel
PY - 2019
Y1 - 2019
N2 - 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.
AB - 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.
KW - High content imaging
KW - High-throughput toxicity screening
KW - Mechanistic safety assessment
KW - Metabolic reprogramming
KW - Mitotoxicity
KW - Neurotoxicity
U2 - 10.1007/s00204-019-02473-y
DO - 10.1007/s00204-019-02473-y
M3 - Journal article
VL - 93
SP - 1585
EP - 1608
JO - Archives of Toxicology
JF - Archives of Toxicology
SN - 0340-5761
IS - 6
ER -