EURL ECVAM strategy to avoid and reduce animal use in genotoxicity testing

The assessment of genotoxicity represents an important component of the safety assessment of all types of substances. Although several in vitro tests are available at different stages of development and acceptance, they cannot at present be considered to fully replace animal tests needed to evaluate the safety of substances. Based on an analysis of regulatory requirements for this endpoint within different pieces of EU legislation, EURL ECVAM proposes a pragmatic approach to improve the traditional genotoxicity testing paradigm that offers solutions in both the shortand medium-term and that draws on the considerable experience of 40 years of regulatory toxicology testing in this area. EURL ECVAM considers that efforts should be directed towards the overall improvement of the current testing strategy for better hazard and risk assessment approaches, which either avoids or minimises the use of animals, whilst satisfying regulatory information requirements, irrespective of regulatory context. Several opportunities for the improvement of the testing strategy have been identified which aim to i) enhance the performance of the in vitro testing battery so that fewer in vivo follow-up tests are necessary and ii) guide more intelligent in vivo follow-up testing to reduce unnecessary use of animals. The implementation of this strategic plan will rely on the cooperation of EURL ECVAM with other existing initiatives and the coordinated contribution from various stakeholders. As the Commission’s in-house science service, the Joint Research Centre’s mission is to provide EU policies with independent, evidence-based scientific and technical support throughout the whole policy cycle. Working in close cooperation with policy Directorates-General, the JRC addresses key societal challenges while stimulating innovation through developing new standards, methods and tools, and sharing and transferring its know-how to the Member States and international community. Key policy areas include: environment and climate change; energy and transport; agriculture and food security; health and consumer protection; information society and digital agenda; safety and security including nuclear; all supported through a cross- cutting and multi-disciplinary approach. L B -N A -2 6 3 7 5 -E N -N

[1]  Raffaella Corvi,et al.  ECVAM prevalidation of three cell transformation assays. , 2011, ALTEX.

[2]  Lapenna Silvia,et al.  The Applicability of Software Tools for Genotoxicity and Carcinogenicity Prediction: Case Studies relevant to the Assessment of Pesticides , 2010 .

[3]  A. Gennery,et al.  Primary immunodeficiencies associated with DNA-repair disorders , 2010, Expert Reviews in Molecular Medicine.

[4]  J. Kleinjans,et al.  Application of toxicogenomics to study mechanisms of genotoxicity and carcinogenicity. , 2009, Toxicology letters.

[5]  Sunanda Kadam,et al.  Globally Harmonized System of Classification and Labeling of Chemicals , 2010 .

[6]  Andreas Czich,et al.  Follow‐up actions from positive results of in vitro genetic toxicity testing , 2011, Environmental and molecular mutagenesis.

[7]  M. Waters,et al.  Characterizing and predicting carcinogenicity and mode of action using conventional and toxicogenomics methods. , 2010, Mutation research.

[8]  Andrew Williams,et al.  Global transcriptional characterization of a mouse pulmonary epithelial cell line for use in genetic toxicology. , 2009, Toxicology in vitro : an international journal published in association with BIBRA.

[9]  Emilio Benfenati,et al.  Comparison of In Silico Models for Prediction of Mutagenicity , 2013, Journal of environmental science and health. Part C, Environmental carcinogenesis & ecotoxicology reviews.

[10]  Jiri Aubrecht,et al.  New and emerging technologies for genetic toxicity testing , 2011, Environmental and molecular mutagenesis.

[11]  Raffaella Corvi,et al.  Analysis of published data for top concentration considerations in mammalian cell genotoxicity testing. , 2010, Mutagenesis.

[12]  S. Frank.  Somatic evolutionary genomics: Mutations during development cause highly variable genetic mosaicism with risk of cancer and neurodegeneration , 2010, Proceedings of the National Academy of Sciences.

[13]  A. Elhajouji,et al.  In vitro genotoxicity test approaches with better predictivity: summary of an IWGT workshop. , 2011, Mutation research.

[14]  Ruili Huang,et al.  Characterization of environmental chemicals with potential for DNA damage using isogenic DNA repair‐deficient chicken DT40 cell lines , 2011, Environmental and molecular mutagenesis.

[15]  R. Erickson.  Somatic gene mutation and human disease other than cancer: an update. , 2010, Mutation research.

[16]  Munn Sharon j.,et al.  Alternative Approaches Can Reduce the Use of Test Animals under REACH. , 2004 .

[17]  Andrew Worth,et al.  Structural analysis and predictive value of the rodent in vivo micronucleus assay results. , 2010, Mutagenesis.

[18]  Andreas Czich,et al.  Collaborative study on fifteen compounds in the rat-liver Comet assay integrated into 2- and 4-week repeat-dose studies. , 2010, Mutation research.

[19]  N. Banduhn,et al.  The hen's egg test for micronucleus induction (HET-MN): novel analyses with a series of well-characterized substances support the further evaluation of the test system. , 2008, Mutation research.

[20]  D. Kirkland,et al.  Evaluation of a multi-endpoint assay in rats, combining the bone-marrow micronucleus test, the Comet assay and the flow-cytometric peripheral blood micronucleus test. , 2011, Mutation research.

[21]  David Kirkland,et al.  Further analysis of Ames-negative rodent carcinogens that are only genotoxic in mammalian cells in vitro at concentrations exceeding 1 mM, including retesting of compounds of concern. , 2010, Mutagenesis.

[22]  T. Singer,et al.  Comparative evaluation of in silico systems for ames test mutagenicity prediction: scope and limitations. , 2011, Chemical research in toxicology.

[23]  Nicola J Hewitt,et al.  A tiered approach to the use of alternatives to animal testing for the safety assessment of cosmetics: genotoxicity. A COLIPA analysis. , 2010, Regulatory toxicology and pharmacology : RTP.

[24]  Paul Jennings,et al.  An overview of transcriptional regulation in response to toxicological insult , 2012, Archives of Toxicology.

[25]  N. Tanaka,et al.  An international validation study of a Bhas 42 cell transformation assay for the prediction of chemical carcinogenicity. , 2011, Mutation research.

[26]  Raffaella Corvi,et al.  How to reduce false positive results when undertaking in vitro genotoxicity testing and thus avoid unnecessary follow-up animal tests: Report of an ECVAM Workshop. , 2007, Mutation research.

[27]  J. Hoeijmakers.  DNA damage, aging, and cancer. , 2009, The New England journal of medicine.

[28]  W. Schoonen,et al.  The development of RAD51C, Cystatin A, p53 and Nrf2 luciferase-reporter assays in metabolically competent HepG2 cells for the assessment of mechanism-based genotoxicity and of oxidative stress in the early research phase of drug development. , 2010, Mutation research.

[29]  Vera Rogiers,et al.  Opportunities for an alternative integrating testing strategy for carcinogen hazard assessment? , 2012, Critical reviews in toxicology.

[30]  R L Binder,et al.  Xenobiotic metabolism gene expression in the EpiDermin vitro 3D human epidermis model compared to human skin. , 2010, Toxicology in vitro : an international journal published in association with BIBRA.

[31]  Raffaella Corvi,et al.  ECVAM retrospective validation of in vitro micronucleus test (MNT) , 2008, Mutagenesis.

[32]  Raffaella Corvi,et al.  Reduction of use of animals in regulatory genotoxicity testing: Identification and implementation opportunities-Report from an ECVAM workshop. , 2009, Mutation research.

[33]  J. Kleinjans,et al.  Transcriptomic responses generated by hepatocarcinogens in a battery of liver-based in vitro models. , 2013, Carcinogenesis.

[34]  Paul Cahill,et al.  GADD45a-GFP GreenScreen HC assay results for the ECVAM recommended lists of genotoxic and non-genotoxic chemicals for assessment of new genotoxicity tests. , 2010, Mutation research.

[35]  David Kirkland,et al.  A core in vitro genotoxicity battery comprising the Ames test plus the in vitro micronucleus test is sufficient to detect rodent carcinogens and in vivo genotoxins. , 2011, Mutation research.

[36]  Bas J Blaauboer,et al.  An expert consortium review of the EC-commissioned report "alternative (Non-Animal) methods for cosmetics testing: current status and future prospects - 2010". , 2011, ALTEX.

[37]  Romualdo Benigni,et al.  Investigating the relationship between in vitro-in vivo genotoxicity: derivation of mechanistic QSAR models for in vivo liver genotoxicity and in vivo bone marrow micronucleus formation which encompass metabolism. , 2012, Chemical research in toxicology.

[38]  P. White,et al.  Proliferating primary hepatocytes from the pUR288 lacZ plasmid mouse are valuable tools for genotoxicity assessment in vitro , 2012, Environmental and molecular mutagenesis.

[39]  S. De Flora,et al.  Mutagenesis and cardiovascular diseases Molecular mechanisms, risk factors, and protective factors. , 2007, Mutation research.

[40]  R. Corvi,et al.  International prevalidation study on cell transformation assay. Preface. , 2012, Mutation research.

[41]  Olga Tcheremenskaia,et al.  The new ISSMIC database on in vivo micronucleus and its role in assessing genotoxicity testing strategies. , 2012, Mutagenesis.

[42]  Raffaella Corvi,et al.  Comet assay in reconstructed 3D human epidermal skin models—investigation of intra- and inter-laboratory reproducibility with coded chemicals , 2013, Mutagenesis.

[43]  Thomas Hartung,et al.  Meeting Report: Validation of Toxicogenomics-Based Test Systems: ECVAM–ICCVAM/NICEATM Considerations for Regulatory Use , 2005, Environmental health perspectives.

[44]  Katie Smith,et al.  Reduction of misleading ("false") positive results in mammalian cell genotoxicity assays. I. Choice of cell type. , 2012, Mutation research.

[45]  Kerstin Reisinger,et al.  Applicability and robustness of the hen's egg test for analysis of micronucleus induction (HET-MN): results from an inter-laboratory trial. , 2012, Mutation research.

[46]  Raffaella Corvi,et al.  Recommended lists of genotoxic and non-genotoxic chemicals for assessment of the performance of new or improved genotoxicity tests: a follow-up to an ECVAM workshop. , 2008, Mutation research.

[47]  Nicola J Hewitt,et al.  International prevalidation studies of the EpiDerm 3D human reconstructed skin micronucleus (RSMN) assay: transferability and reproducibility. , 2010, Mutation research.

[48]  David Kirkland,et al.  Evaluation of the ability of a battery of three in vitro genotoxicity tests to discriminate rodent carcinogens and non-carcinogens III. Appropriate follow-up testing in vivo. , 2005, Mutation research.

[49]  Richard M Walmsley,et al.  High-specificity and high-sensitivity genotoxicity assessment in a human cell line: validation of the GreenScreen HC GADD45a-GFP genotoxicity assay. , 2006, Mutation research.

[50]  M. Aardema,et al.  Use of Syrian hamster embryo and BALB/c 3T3 cell transformation for assessing the carcinogenic potential of chemicals. , 1999, IARC scientific publications.

[51]  Raffaella Corvi,et al.  3.7. Genotoxicity and Mutagenicity , 2005 .

[52]  Hyung Sik Kim,et al.  Mutagenicity and carcinogenicity: human reproductive cancer and risk factors , 2011 .

[53]  Katie Smith,et al.  Reduction of misleading ("false") positive results in mammalian cell genotoxicity assays. II. Importance of accurate toxicity measurement. , 2012, Mutation research.

[54]  Leslie Recio,et al.  Improvement of in vivo genotoxicity assessment: combination of acute tests and integration into standard toxicity testing. , 2011, Mutation research.