Genotoxic mode of action predictions from a multiplexed flow cytometric assay and a machine learning approach

Environmental and Molecular Mutagenesis
Steven M BryceStephen D Dertinger

Abstract

Several endpoints associated with cellular responses to DNA damage as well as overt cytotoxicity were multiplexed into a miniaturized, "add and read" type flow cytometric assay. Reagents included a detergent to liberate nuclei, RNase and propidium iodide to serve as a pan-DNA dye, fluorescent antibodies against γH2AX, phospho-histone H3, and p53, and fluorescent microspheres for absolute nuclei counts. The assay was applied to TK6 cells and 67 diverse reference chemicals that served as a training set. Exposure was for 24 hrs in 96-well plates, and unless precipitation or foreknowledge about cytotoxicity suggested otherwise, the highest concentration was 1 mM. At 4- and 24-hrs aliquots were removed and added to microtiter plates containing the reagent mix. Following a brief incubation period robotic sampling facilitated walk-away data acquisition. Univariate analyses identified biomarkers and time points that were valuable for classifying agents into one of three groups: clastogenic, aneugenic, or non-genotoxic. These mode of action predictions were optimized with a forward-stepping process that considered Wald test p-values, receiver operator characteristic curves, and pseudo R(2) values, among others. A particularly high perfo...Continue Reading

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Citations

Jul 2, 2016·Environmental and Molecular Mutagenesis·Derek T BernackiStephen D Dertinger
Oct 22, 2016·Chemical Research in Toxicology·Mikael Persson, Jorrit J Hornberg
Aug 25, 2016·Food and Chemical Toxicology : an International Journal Published for the British Industrial Biological Research Association·Raffaella Corvi, Federica Madia
Apr 19, 2018·Environmental and Molecular Mutagenesis·Stephanie L Smith-RoeKristine L Witt
Nov 7, 2017·Toxicological Sciences : an Official Journal of the Society of Toxicology·Steven M BryceStephen D Dertinger
May 28, 2019·Toxicological Sciences : an Official Journal of the Society of Toxicology·Derek T BernackiStephen D Dertinger
Oct 1, 2019·Environmental and Molecular Mutagenesis·Derek T BernackiStephanie L Smith-Roe
Feb 11, 2020·Environmental and Molecular Mutagenesis·Shuchang TianStephen D Dertinger
Jul 11, 2019·Archives of Toxicology·B KoppMarc Audebert
Apr 4, 2017·Environmental and Molecular Mutagenesis·Steven M BryceStephen D Dertinger
Oct 31, 2019·Archives of Toxicology·Shuaizhang Li, Menghang Xia
Nov 1, 2017·Environmental Science and Pollution Research International·Jamile Silveira TomiazziAna Paula Alves Favareto
Jun 25, 2017·Environmental and Molecular Mutagenesis·Stephen D Dertinger
Sep 29, 2020·Journal of Toxicology and Environmental Health. Part a·Xiaoqing GuoNan Mei
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Jul 4, 2020·Toxicological Sciences : an Official Journal of the Society of Toxicology·Inger BrandsmaGiel Hendriks
Jan 12, 2021·Environmental and Molecular Mutagenesis·Ryan P WheeldonGeorge E Johnson
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Apr 9, 2020·Mutation Research. Genetic Toxicology and Environmental Mutagenesis·Ludovic Le HegaratValérie Fessard
Apr 9, 2020·Mutation Research. Genetic Toxicology and Environmental Mutagenesis·Laure KhouryMarc Audebert

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