ToxTracker Suite

The ToxTracker suite contains a unique collection of in vitro genotoxicity assays that can reliably identify genotoxic compounds and can provide detailed insight into their mode-of-action. ToxTracker consists of a collection of 6 unique fluorescent reporter cell lines that are combined in a single assay. These reporters signal the activation of various cellular stress responses that indicate the induction of DNA damage, oxidative stress, and protein damage. The extended assay, ToxTracker ACEcombines these different fluorescent reporters with an extensive cell cycle analysis and polyploidy analysis to discriminate between genotoxins with a clastogenic or an aneugenic mode-of-action. Aneugens can be further characterised in TubulinTracker to support an Adverse Outcome Pathway (AOP) approach. Genotoxic effects that are indirectly caused by high levels of oxidative stress can be readily identified in ToxTracker AO. The ToxTracker suite is particularly useful for AOP and read-across approaches. All assays can be useful as stand-alone or as follow-up tests of ToxTracker.

ToxTracker is typically used for early genotoxicity screening and as a follow-up test for mode-of-action assessment. ToxTracker requires small amounts of test material, has a short turn-around time, and has an excellent predictivity for the Ames mutation test, as well as, the in vitro and in vivo micronucleus assay. The mode-of-action information from the ToxTracker suite can be highly valuable during compound development or in a weight-of-evidence approach for regulatory application.

ToxTracker is a stem cell-based assay that combines 6 fluorescent reporter cell lines to discriminate between induction of DNA damage, induction of oxidative stress, and protein damage. Their collective induction profile provides mechanistic insight into specific genotoxic properties of compounds.

Typical applications

  • Identification of genotoxic compounds with high sensitivity and specificity
  • High throughput genotoxicity screening
  • Investigations on mode-of-action of genotoxic compounds

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ToxTracker ACE (Aneugen and Clastogen Evaluation) is an extended version of the ToxTracker assay and includes cell cycle analysis and polyploidy detection in addition to activation of the 6 fluorescent ToxTracker reporters to allow direct discrimination between an aneugenic and clastogenic mode-of-action of genotoxic compounds. ToxTracker ACE is able to separate clastogens from aneugens and identify tubulin poisons and cell cycle kinase inhibitors.

Typical applications

  • Genotoxicity testing of pharmaceutical compounds
  • Identification of aneugens
  • Genotoxicity testing of kinase inhibitors

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TubulinTracker is an extension to the collection of fluorescent reporter cell lines in ToxTracker. The TubulinTracker assay is a tubulin-GFP reporter assay that can be applied to visualise microtubule polymerisation dynamics, progression of cells through mitosis, and aneugenicity. TubulinTracker has been developed as a follow-up for the ToxTracker ACE assay to further assess the mode of action of aneugenic compounds by discriminating between tubulin stabilisers and destabilisers.

Typical applications

  • Follow-up of ToxTracker (ACE) for compounds with an aneugenic mode-of-action
  • Follow-up of a positive in vitro micronucleus test
  • AOP approach for tubulin poisons

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High levels of reactive oxygen species (ROS) and oxidative stress can lead to DNA damage. To investigate the contribution of oxidative stress in the genotoxic profile of a compound, ToxTracker can be performed in presence of a ROS scavenger (NAC and GSH) and measure its impact on DNA damage, oxidative stress, and cytotoxicity. This will allow for the discrimination between direct DNA reactivity of a compound and indirect genotoxicity caused by oxidative stress.

Typical applications

  • Genotoxicity testing of compounds when oxidative stress is the expected mode-of-action
  • Follow-up of a positive ToxTracker test 
  • Follow-up of a positive in vitro micronucleus test

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ToxPlot is a unique software for automated ToxTracker data analysis and unbiased data interpretation. Graphical representation of the test results allows for a clear and rapid assessment of the reactive properties of compounds. Data analysis is customisable and reports can be easily generated with just the push of a button. ToxPlot is available for every ToxTracker project that is performed by Toxys as a service as well as for projects that are performed in your own lab.

Typical applications

  • Data analysis for the ToxTracker assay
  • Automated report generation
  • Building a database with all your historical ToxTracker results
  • Compare test results from different projects and compare them with a library of reference compounds

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