New and emerging technologies for genetic toxicity testing

The International Life Sciences Institute (ILSI) Health and Environmental Sciences Institute (HESI) Project Committee on the Relevance and Follow‐up of Positive Results in In Vitro Genetic Toxicity (IVGT) Testing established an Emerging Technologies and New Strategies Workgroup to review the current State of the Art in genetic toxicology testing. The aim of the workgroup was to identify promising technologies that will improve genotoxicity testing and assessment of in vivo hazard and risk, and that have the potential to help meet the objectives of the IVGT. As part of this initiative, HESI convened a workshop in Washington, DC in May 2008 to discuss mature, maturing, and emerging technologies in genetic toxicology. This article collates the abstracts of the New and Emerging Technologies Workshop together with some additional technologies subsequently considered by the workgroup. Each abstract (available in the online version of the article) includes a section addressed specifically to the strengths, weaknesses, opportunities, and threats associated with the respective technology. Importantly, an overview of the technologies and an indication of how their use might be aligned with the objectives of IVGT are presented. In particular, consideration was given with regard to follow‐up testing of positive results in the standard IVGT tests (i.e., Salmonella Ames test, chromosome aberration assay, and mouse lymphoma assay) to add weight of evidence and/or provide mechanism of action for improved genetic toxicity risk assessments in humans. Environ. Mol. Mutagen., 2011. © 2010 Wiley‐Liss, Inc.

[1]  J. Heddle,et al.  The production of micronuclei from chromosome aberrations in irradiated cultures of human lymphocytes. , 1976, Mutation research.

[2]  D. Richardson,et al.  Growth arrest and DNA damage-45 alpha (GADD45alpha). , 2009, The international journal of biochemistry & cell biology.

[3]  Flamand Nicole,et al.  再構築ヒト皮膚モデル,Episkinによる遺伝毒性試験手順の開発:経皮適用化合物のin vitroでの危険性評価のための新しいツールか? , 2006 .

[4]  R. Schiestl,et al.  The aromatic amine carcinogens o-toluidine and o-anisidine induce free radicals and intrachromosomal recombination in Saccharomyces cerevisiae. , 1999, Mutation research.

[5]  Wolfgang Muster,et al.  Computational toxicology in drug development. , 2008, Drug discovery today.

[6]  Jiri Aubrecht,et al.  Toxicogenomics: overview and potential applications for the study of non-covalent DNA interacting chemicals. , 2007, Mutation research.

[7]  Antonio Marcomini,et al.  Genotoxicity, cytotoxicity, and reactive oxygen species induced by single‐walled carbon nanotubes and C60 fullerenes in the FE1‐Muta™Mouse lung epithelial cells , 2008, Environmental and molecular mutagenesis.

[8]  Martin M. Schumacher,et al.  Use of the alkaline in vivo Comet assay for mechanistic genotoxicity investigations. , 2004, Mutagenesis.

[9]  M. Bergqvist,et al.  Human cytogenetic biomonitoring using flow‐cytometric analysis of micronuclei in transferrin‐positive immature peripheral blood reticulocytes , 2000, Environmental and molecular mutagenesis.

[10]  S. Dertinger,et al.  Practical threshold for micronucleated reticulocyte induction observed for low doses of mitomycin C, Ara-C and colchicine. , 2006, Mutagenesis.

[11]  T. Sofuni Japanese guidelines for mutagenicity testing , 1993 .

[12]  J. T. Macgregor,et al.  Flow cytometric analysis of micronuclei in peripheral blood reticulocytes III. An efficient method of monitoring chromosomal damage in the beagle dog. , 2007, Toxicological sciences : an official journal of the Society of Toxicology.

[13]  R. Mittelstaedt,et al.  Development of an in vivo gene mutation assay using the endogenous Pig‐A gene: II. Selection of Pig‐A mutant rat spleen T‐cells with proaerolysin and sequencing Pig‐A cDNA from the mutants , 2008, Environmental and molecular mutagenesis.

[14]  Laurent Griscom,et al.  Trends in the development of microfluidic cell biochips for in vitro hepatotoxicity. , 2007, Toxicology in vitro : an international journal published in association with BIBRA.

[15]  U. Vogel,et al.  Diesel exhaust particles are mutagenic in FE1-MutaMouse lung epithelial cells. , 2008, Mutation research.

[16]  S. Dertinger,et al.  Analysis of micronucleated cells by flow cytometry. 1. Achieving high resolution with a malaria model. , 1993, Mutation research.

[17]  Stephen D Dertinger,et al.  Interlaboratory evaluation of a flow cytometric, high content in vitro micronucleus assay. , 2008, Mutation research.

[18]  Daniel Marzin,et al.  SFTG international collaborative study on in vitro micronucleus test I. General conditions and overall conclusions of the study. , 2006, Mutation research.

[19]  David Kirkland,et al.  SFTG international collaborative study on in vitro micronucleus test , 2006 .

[20]  H. Sugimura,et al.  Development of the adductome approach to detect DNA damage in humans. , 2006, Antioxidants & redox signaling.

[21]  J. Schenkman,et al.  Biochemical applications of ultrathin films of enzymes, polyions and DNA. , 2008, Chemical communications.

[22]  J. Bemis,et al.  High content flow cytometric micronucleus scoring method is applicable to attachment cell lines , 2009, Environmental and molecular mutagenesis.

[23]  R. Tice,et al.  Single cell gel/comet assay: Guidelines for in vitro and in vivo genetic toxicology testing , 2000, Environmental and molecular mutagenesis.

[24]  R. Walmsley,et al.  Interlaboratory assessment of the GreenScreen HC GADD45a-GFP genotoxicity screening assay: an enabling study for independent validation as an alternative method. , 2008, Mutation research.

[25]  B. Vogelstein,et al.  A mammalian cell cycle checkpoint pathway utilizing p53 and GADD45 is defective in ataxia-telangiectasia , 1992, Cell.

[26]  Yuhchyau Chen,et al.  Automated human blood micronucleated reticulocyte measurements for rapid assessment of chromosomal damage. , 2007, Mutation research.

[27]  J. T. Macgregor,et al.  Flow cytometric analysis of micronuclei in peripheral blood reticulocytes: I. Intra- and interlaboratory comparison with microscopic scoring. , 2006, Toxicological sciences : an official journal of the Society of Toxicology.

[28]  K. Witt,et al.  Elevated frequencies of micronucleated erythrocytes in infants exposed to zidovudine in utero and postpartum to prevent mother‐to‐child transmission of HIV , 2007, Environmental and molecular mutagenesis.

[29]  B. Bhaskar Gollapudi,et al.  Genetic Toxicity Assessment: Employing the Best Science for Human Safety Evaluation , 2006 .

[30]  Nigel Greene,et al.  Computer systems for the prediction of toxicity: an update. , 2002, Advanced drug delivery reviews.

[31]  J. Cadet,et al.  Creating context for the use of DNA adduct data in cancer risk assessment: II. Overview of methods of identification and quantitation of DNA damage , 2009, Critical reviews in toxicology.

[32]  P. White,et al.  Development and characterization of a stable epithelial cell line from Muta™Mouse lung , 2003, Environmental and molecular mutagenesis.

[33]  N. Kruhlak,et al.  An analysis of genetic toxicity, reproductive and developmental toxicity, and carcinogenicity data: II. Identification of genotoxicants, reprotoxicants, and carcinogens using in silico methods. , 2006, Regulatory toxicology and pharmacology : RTP.

[34]  M. Fenech,et al.  Report from the In Vitro Micronucleus Assay Working Group , 2003, Mutation research.

[35]  Raymond R Tice,et al.  Fourth International Workgroup on Genotoxicity testing: results of the in vivo Comet assay workgroup. , 2007, Mutation research.

[36]  R. Tice,et al.  A simple technique for quantitation of low levels of DNA damage in individual cells. , 1988, Experimental cell research.

[37]  S. Stasiewicz,et al.  Chemicals showing no evidence of carcinogenicity in long-term, two-species rodent studies: the need for short-term test data. , 1984, Environmental mutagenesis.

[38]  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.

[39]  Marilyn J Aardema,et al.  Identification of a gene expression profile that discriminates indirect-acting genotoxins from direct-acting genotoxins. , 2004, Mutation research.

[40]  A. Fornace,et al.  Genetic toxicity assessment: employing the best science for human safety evaluation Part VII: Why not start with a single test: a transformational alternative to genotoxicity hazard and risk assessment. , 2007, Toxicological sciences : an official journal of the Society of Toxicology.

[41]  R. Schiestl,et al.  Oxidative mutagens induce intrachromosomal recombination in yeast. , 1994, Mutation research.

[42]  M. Cimino,et al.  Genotoxicity risk assessment: a proposed classification strategy. , 2002, Mutation research.

[43]  C. Reiners,et al.  Evaluation of the reticulocyte micronucleus assay in patients treated with radioiodine for thyroid cancer. , 2005, Mutation research.

[44]  Marilyn J Aardema,et al.  Development of a method for assessing micronucleus induction in a 3D human skin model (EpiDerm). , 2006, Mutation research.

[45]  Barbara Hoffman,et al.  Gadd45 in the response of hematopoietic cells to genotoxic stress. , 2007, Blood cells, molecules & diseases.

[46]  Lutz Müller,et al.  Evaluation of the ability of a battery of three in vitro genotoxicity tests to discriminate rodent carcinogens and non-carcinogens II. Further analysis of mammalian cell results, relative predictivity and tumour profiles. , 2006, Mutation research.

[47]  Richard M Walmsley,et al.  Assessment of the genotoxicity of S9-generated metabolites using the GreenScreen HC GADD45a-GFP assay. , 2008, Mutagenesis.

[48]  Akio Yamada,et al.  OECD Guidelines for Testing of Chemicals , 1982 .

[49]  J. Aubrecht,et al.  In vivo DNA deletion assay to detect environmental and genetic predisposition to cancer. , 2004, Methods in molecular biology.

[50]  A. Knaap,et al.  Role of genotoxicity assays in the regulation of chemicals in The Netherlands: considerations and experiences. , 1991, Mutagenesis.

[51]  Errol Zeiger,et al.  Creating context for the use of DNA adduct data in cancer risk assessment: I. Data organization , 2009, Critical reviews in toxicology.

[52]  R. Brodsky,et al.  PIG-A mutations in paroxysmal nocturnal hemoglobinuria and in normal hematopoiesis , 2006, Leukemia & lymphoma.

[53]  J. T. Macgregor,et al.  Integration of mutation and chromosomal damage endpoints into 28-day repeat dose toxicology studies. , 2010, Toxicological sciences : an official journal of the Society of Toxicology.

[54]  John C. Dearden,et al.  In silico prediction of drug toxicity , 2003, J. Comput. Aided Mol. Des..

[55]  Jiri Aubrecht,et al.  Gene expression profile analysis: an emerging approach to investigate mechanisms of genotoxicity. , 2005, Pharmacogenomics.

[56]  M. Cimino,et al.  Comparative overview of current international strategies and guidelines for genetic toxicology testing for regulatory purposes , 2006, Environmental and molecular mutagenesis.

[57]  Stephen D Dertinger,et al.  Miniaturized flow cytometric in vitro micronucleus assay represents an efficient tool for comprehensively characterizing genotoxicity dose-response relationships. , 2010, Mutation research.

[58]  M. Aardema,et al.  Intralaboratory and interlaboratory evaluation of the EpiDerm 3D human reconstructed skin micronucleus (RSMN) assay. , 2009, Mutation research.

[59]  Nicklas Raun Jacobsen,et al.  Increased mutant frequency by carbon black, but not quartz, in the lacZ and cII transgenes of muta™mouse lung epithelial cells , 2007, Environmental and molecular mutagenesis.

[60]  R. Heflich,et al.  Accumulation and persistence of Pig-A mutant peripheral red blood cells following treatment of rats with single and split doses of N-ethyl-N-nitrosourea. , 2009, Mutation research.

[61]  Mitchell Klausner,et al.  Further development of the EpiDerm 3D reconstructed human skin micronucleus (RSMN) assay. , 2009, Mutation research.

[62]  Jing Shi,et al.  Further evaluation of a flow cytometric in vitro micronucleus assay in CHO-K1 cells: a reliable platform that detects micronuclei and discriminates apoptotic bodies. , 2010, Mutagenesis.

[63]  Peter Kasper,et al.  Relevance and follow-up of positive results in in vitro genetic toxicity assays: an ILSI-HESI initiative. , 2007, Mutation research.

[64]  J. Schenkman,et al.  Toxicity screening using biosensors that measure DNA damage. , 2007, Current opinion in drug discovery & development.

[65]  Véronique Thybaud,et al.  Application of toxicogenomics to genetic toxicology risk assessment , 2007, Environmental and molecular mutagenesis.

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

[67]  P. Harbach,et al.  Interlaboratory validation of a CD71‐based flow cytometric method (Microflow®) for the scoring of micronucleated reticulocytes in mouse peripheral blood , 2005, Environmental and molecular mutagenesis.

[68]  M. Nüsse,et al.  Flow cytometric analysis of micronuclei found in cells after irradiation. , 1984, Cytometry.

[69]  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.

[70]  L. Marrot,et al.  Development of genotoxicity test procedures with Episkin, a reconstructed human skin model: towards new tools for in vitro risk assessment of dermally applied compounds? , 2006, Mutation research.

[71]  R. Walmsley,et al.  Genotoxic carcinogen or not genotoxic carcinogen? That is the question Re: Olaharski et al., "Validation of the GreenScreen GADD45alpha-GFP indicator assay with non-proprietary and proprietary compounds". , 2009, Mutation research.

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

[73]  Stephen D Dertinger,et al.  Comparative scoring of micronucleated reticulocytes in rat peripheral blood by flow cytometry and microscopy. , 2003, Toxicological sciences : an official journal of the Society of Toxicology.

[74]  A. Sarasin,et al.  Characterization of DNA reactive and non-DNA reactive anticancer drugs by gene expression profiling. , 2007, Mutation research.

[75]  B H Margolin,et al.  Prediction of chemical carcinogenicity in rodents from in vitro genetic toxicity assays. , 1987, Science.

[76]  N. Kruhlak,et al.  An analysis of genetic toxicity, reproductive and developmental toxicity, and carcinogenicity data: I. Identification of carcinogens using surrogate endpoints. , 2006, Regulatory toxicology and pharmacology : RTP.

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

[78]  A. Vickers,et al.  Precision-cut organ slices to investigate target organ injury , 2005, Expert opinion on drug metabolism & toxicology.

[79]  G. Warnes,et al.  Differentiation of DNA reactive and non-reactive genotoxic mechanisms using gene expression profile analysis. , 2004, Mutation research.

[80]  E Zeiger,et al.  Strategy for genotoxicity testing: hazard identification and risk assessment in relation to in vitro testing. , 2007, Mutation research.

[81]  R. Brennan,et al.  Detecting carcinogens with the yeast DEL assay. , 2004, Methods in molecular biology.

[82]  Jiri Aubrecht,et al.  The utility of DNA microarrays for characterizing genotoxicity. , 2004, Environmental health perspectives.

[83]  Or Guidance for Industry S 2 ( R 1 ) Genotoxicity Testing and Data Interpretation for Pharmaceuticals Intended for Human Use , 2012 .

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

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

[86]  J. T. Macgregor,et al.  Sensitivity of the erythrocyte micronucleus assay: dependence on number of cells scored and inter-animal variability. , 2007, Mutation research.

[87]  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.

[88]  Richard M Walmsley,et al.  Development and Validation of a Higher Throughput Screening Approach to Genotoxicity Testing Using the GADD45a-GFP GreenScreen HC Assay , 2009, Journal of biomolecular screening.

[89]  E. Zeiger Mutagens that are not carcinogens: faulty theory or faulty tests? , 2001, Mutation research.

[90]  Trevelyan Square,et al.  Recommendations for conducting the in vivo alkaline Comet assay , 2003 .

[91]  Y. Katsuura,et al.  Development of an in vivo gene mutation assay using the endogenous Pig‐A gene: I. Flow cytometric detection of CD59‐negative peripheral red blood cells and CD48‐negative spleen T‐cells from the rat , 2008, Environmental and molecular mutagenesis.

[92]  A. Lynch,et al.  Evaluation of the Litron In Vitro MicroFlow Kit for the flow cytometric enumeration of micronuclei (MN) in mammalian cells. , 2008, Mutation research.

[93]  S. Dertinger,et al.  Quantitative analysis of Howell-Jolly bodies in children with sickle cell disease. , 2004, Experimental hematology.

[94]  J. Bemis,et al.  In vivo mutation assay based on the endogenous Pig‐a locus , 2008, Environmental and molecular mutagenesis.

[95]  K J Johanson,et al.  Microelectrophoretic study of radiation-induced DNA damages in individual mammalian cells. , 1984, Biochemical and biophysical research communications.

[96]  J W Green,et al.  A review of the genotoxicity of marketed pharmaceuticals. , 2001, Mutation research.

[97]  Ronald D Snyder,et al.  SFTG international collaborative study on in vitro micronucleus test III. Using CHO cells. , 2006, Mutation research.

[98]  Ollivier Hyrien,et al.  Pig-a mutation: kinetics in rat erythrocytes following exposure to five prototypical mutagens. , 2010, Toxicological sciences : an official journal of the Society of Toxicology.

[99]  Michael L Bittner,et al.  Stress-specific signatures: expression profiling of p53 wild-type and -null human cells , 2005, Oncogene.

[100]  S. Dertinger,et al.  Analysis of micronucleated cells by flow cytometry. 4. Kinetic analysis of cytogenetic damage in blood. , 1995, Mutation research.

[101]  S. Dertinger,et al.  Enumeration of micronucleated reticulocytes in rat peripheral blood: a flow cytometric study. , 2000, Mutation research.

[102]  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.

[103]  D. L. Grant,et al.  The assessment of mutagenicity health protection branch mutagenicity guidelines , 1993, Environmental and molecular mutagenesis.

[104]  Richard M Walmsley,et al.  Analysis of 75 marketed pharmaceuticals using the GADD45a-GFP 'GreenScreen HC' genotoxicity assay. , 2009, Mutagenesis.

[105]  S. Bremer,et al.  The possible impact of human embryonic stem cells on safety pharmacological and toxicological assessments in drug discovery and drug development. , 2008, Current stem cell research & therapy.

[106]  Gunnar Brunborg,et al.  The comet assay: topical issues. , 2008, Mutagenesis.

[107]  Q. Zhan Gadd45a, a p53- and BRCA1-regulated stress protein, in cellular response to DNA damage. , 2005, Mutation research.

[108]  GUIDANCE DOCUMENT,et al.  Guidance on Genotoxicity Testing and Data Interpretation for Pharmaceuticals Intended for Human Use , 2008 .

[109]  B. Vessby,et al.  The impact of folate status and folic acid supplementation on the micronucleus frequency in human erythrocytes. , 2006, Mutation research.

[110]  Stephen D Dertinger,et al.  In vitro micronucleus scoring by flow cytometry: Differential staining of micronuclei versus apoptotic and necrotic chromatin enhances assay reliability , 2006, Environmental and molecular mutagenesis.

[111]  Junichiro Saito,et al.  Flow cytometric analysis of micronuclei in peripheral blood reticulocytes: II. An efficient method of monitoring chromosomal damage in the rat. , 2006, Toxicological sciences : an official journal of the Society of Toxicology.

[112]  Silvio Albertini,et al.  Evaluation of the GreenScreen GADD45alpha-GFP indicator assay with non-proprietary and proprietary compounds. , 2009, Mutation research.