Next generation testing strategy for assessment of genomic damage: A conceptual framework and considerations

For several decades, regulatory testing schemes for genetic damage have been standardized where the tests being utilized examined mutations and structural and numerical chromosomal damage. This has served the genetic toxicity community well when most of the substances being tested were amenable to such assays. The outcome from this testing is usually a dichotomous (yes/no) evaluation of test results, and in many instances, the information is only used to determine whether a substance has carcinogenic potential or not. Over the same time period, mechanisms and modes of action (MOAs) that elucidate a wider range of genomic damage involved in many adverse health outcomes have been recognized. In addition, a paradigm shift in applied genetic toxicology is moving the field toward a more quantitative dose‐response analysis and point‐of‐departure (PoD) determination with a focus on risks to exposed humans. This is directing emphasis on genomic damage that is likely to induce changes associated with a variety of adverse health outcomes. This paradigm shift is moving the testing emphasis for genetic damage from a hazard identification only evaluation to a more comprehensive risk assessment approach that provides more insightful information for decision makers regarding the potential risk of genetic damage to exposed humans. To enable this broader context for examining genetic damage, a next generation testing strategy needs to take into account a broader, more flexible approach to testing, and ultimately modeling, of genomic damage as it relates to human exposure. This is consistent with the larger risk assessment context being used in regulatory decision making. As presented here, this flexible approach for examining genomic damage focuses on testing for relevant genomic effects that can be, as best as possible, associated with an adverse health effect. The most desired linkage for risk to humans would be changes in loci associated with human diseases, whether in somatic or germ cells. The outline of a flexible approach and associated considerations are presented in a series of nine steps, some of which can occur in parallel, which was developed through a collaborative effort by leading genetic toxicologists from academia, government, and industry through the International Life Sciences Institute (ILSI) Health and Environmental Sciences Institute (HESI) Genetic Toxicology Technical Committee (GTTC). The ultimate goal is to provide quantitative data to model the potential risk levels of substances, which induce genomic damage contributing to human adverse health outcomes. Any good risk assessment begins with asking the appropriate risk management questions in a planning and scoping effort. This step sets up the problem to be addressed (e.g., broadly, does genomic damage need to be addressed, and if so, how to proceed). The next two steps assemble what is known about the problem by building a knowledge base about the substance of concern and developing a rational biological argument for why testing for genomic damage is needed or not. By focusing on the risk management problem and potential genomic damage of concern, the next step of assay(s) selection takes place. The work‐up of the problem during the earlier steps provides the insight to which assays would most likely produce the most meaningful data. This discussion does not detail the wide range of genomic damage tests available, but points to types of testing systems that can be very useful. Once the assays are performed and analyzed, the relevant data sets are selected for modeling potential risk. From this point on, the data are evaluated and modeled as they are for any other toxicology endpoint. Any observed genomic damage/effects (or genetic event(s)) can be modeled via a dose‐response analysis and determination of an estimated PoD. When a quantitative risk analysis is needed for decision making, a parallel exposure assessment effort is performed (exposure assessment is not detailed here as this is not the focus of this discussion; guidelines for this assessment exist elsewhere). Then the PoD for genomic damage is used with the exposure information to develop risk estimations (e.g., using reference dose (RfD), margin of exposure (MOE) approaches) in a risk characterization and presented to risk managers for informing decision making. This approach is applicable now for incorporating genomic damage results into the decision‐making process for assessing potential adverse outcomes in chemically exposed humans and is consistent with the ILSI HESI Risk Assessment in the 21st Century (RISK21) roadmap. This applies to any substance to which humans are exposed, including pharmaceuticals, agricultural products, food additives, and other chemicals. It is time for regulatory bodies to incorporate the broader knowledge and insights provided by genomic damage results into the assessments of risk to more fully understand the potential of adverse outcomes in chemically exposed humans, thus improving the assessment of risk due to genomic damage. The historical use of genomic damage data as a yes/no gateway for possible cancer risk has been too narrowly focused in risk assessment. The recent advances in assaying for and understanding genomic damage, including eventually epigenetic alterations, obviously add a greater wealth of information for determining potential risk to humans. Regulatory bodies need to embrace this paradigm shift from hazard identification to quantitative analysis and to incorporate the wider range of genomic damage in their assessments of risk to humans. The quantitative analyses and methodologies discussed here can be readily applied to genomic damage testing results now. Indeed, with the passage of the recent update to the Toxic Substances Control Act (TSCA) in the US, the new generation testing strategy for genomic damage described here provides a regulatory agency (here the US Environmental Protection Agency (EPA), but suitable for others) a golden opportunity to reexamine the way it addresses risk‐based genomic damage testing (including hazard identification and exposure). Environ. Mol. Mutagen. 58:264–283, 2017. © 2016 The Authors. Environmental and Molecular Mutagenesis Published by Wiley Periodicals, Inc.

[1]  Melvin E. Andersen,et al.  Incorporating New Technologies Into Toxicity Testing and Risk Assessment: Moving From 21st Century Vision to a Data-Driven Framework , 2013, Toxicological sciences : an official journal of the Society of Toxicology.

[2]  G. R. Stuart,et al.  Species-specific differences in hepatic mutant frequency and mutational spectrum among lambda/lacI transgenic rats and mice following exposure to aflatoxin B1. , 1997, Carcinogenesis.

[3]  Richard A Becker,et al.  Proposing a scientific confidence framework to help support the application of adverse outcome pathways for regulatory purposes. , 2015, Regulatory toxicology and pharmacology : RTP.

[4]  M. Anderton,et al.  Epigenetics – relevance to drug safety science , 2012 .

[5]  E. Zeiger,et al.  Derivation of point of departure (PoD) estimates in genetic toxicology studies and their potential applications in risk assessment , 2014, Environmental and molecular mutagenesis.

[6]  T Sofuni,et al.  ICH-harmonised guidances on genotoxicity testing of pharmaceuticals: evolution, reasoning and impact. , 1999, Mutation research.

[7]  B. Schumacher,et al.  Genome maintenance and transcription integrity in aging and disease , 2013, Front. Genet..

[8]  J Craig,et al.  Incorporating New Technologies into Toxicity Testing and Risk Assessment: Moving from 21st Century Vision to a Data-Driven Framework , 2013 .

[9]  Jiri Aubrecht,et al.  Application of the TGx‐28.65 transcriptomic biomarker to classify genotoxic and non‐genotoxic chemicals in human TK6 cells in the presence of rat liver S9 , 2016, Environmental and molecular mutagenesis.

[10]  Ord,et al.  A Review of the Reference Dose and Reference Concentration Processes , 2013 .

[11]  Jon M. Maguire,et al.  Prediction of noncovalent Drug/DNA interaction using computational docking models: Studies with over 1350 launched drugs , 2013, Environmental and molecular mutagenesis.

[12]  A. Boobis,et al.  The Key Events Dose-Response Framework: A Cross-Disciplinary Mode-of-Action Based Approach to Examining Dose-Response and Thresholds , 2009, Critical reviews in food science and nutrition.

[13]  M. Hurles,et al.  The genome-wide effects of ionizing radiation on mutation induction in the mammalian germline , 2015, Nature Communications.

[14]  Ocspp,et al.  The Frank R. Lautenberg Chemical Safety for the 21st Century Act , 2016 .

[15]  W. Kittisupamongkol Two sides of the same coin? , 2010, Singapore medical journal.

[16]  Alan R. Boobis,et al.  IPCS Framework for Analyzing the Relevance of a Noncancer Mode of Action for Humans , 2008, Critical reviews in toxicology.

[17]  Y. Dragan,et al.  What do we need to know prior to thinking about incorporating an epigenetic evaluation into safety assessments? , 2010, Toxicological sciences : an official journal of the Society of Toxicology.

[18]  M. E. Meek,et al.  Development of the adverse outcome pathway “alkylation of DNA in male premeiotic germ cells leading to heritable mutations” using the OECD's users' handbook supplement , 2015, Environmental and molecular mutagenesis.

[19]  E. Zeiger,et al.  Quantitative approaches for assessing dose–response relationships in genetic toxicology studies , 2013, Environmental and molecular mutagenesis.

[20]  M. LeBaron,et al.  Epigenetic screening in product safety assessment: are we there yet? , 2011, Toxicology mechanisms and methods.

[21]  English Only Economic Commission for Europe , 2012 .

[22]  Peter A. Jones,et al.  Cancer genetics and epigenetics: two sides of the same coin? , 2012, Cancer cell.

[23]  G. R. Stuart,et al.  Species-specific differences in hepatic mutant frequency and mutational spectrum among lambda/lacl transgenic rats and mice following exposure to aflatoxin B1 , 1996 .

[24]  C. Yauk,et al.  The development of adverse outcome pathways for mutagenic effects for the organization for economic co‐operation and development , 2013, Environmental and molecular mutagenesis.

[25]  Ord,et al.  Integrated Risk Information System , 2013 .

[26]  Carolyn Vickers,et al.  IPCS framework for analysing the relevance of a cancer mode of action for humans , 2006 .

[27]  Shiu-Nan Chen,et al.  Apoptotic cell: linkage of inflammation and wound healing , 2014, Front. Pharmacol..

[28]  E. Zeiger,et al.  An organizational approach for the assessment of DNA adduct data in risk assessment: case studies for aflatoxin B1, tamoxifen and vinyl chloride , 2014, Critical reviews in toxicology.

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

[30]  Gareth J.S. Jenkins,et al.  New approaches to advance the use of genetic toxicology analyses for human health risk assessment , 2015 .

[31]  David H Phillips,et al.  Mutagenicity testing for chemical risk assessment: update of the WHO/IPCS Harmonized Scheme. , 2009, Mutagenesis.

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

[33]  J. Bailar,et al.  Toxicity Testing in the 21st Century: A Vision and a Strategy , 2010, Journal of toxicology and environmental health. Part B, Critical reviews.

[34]  M. Bennett,et al.  DNA Damage and Repair in Vascular Disease. , 2016, Annual review of physiology.

[35]  C. Ioannides,et al.  Cytochromes P450 and species differences in xenobiotic metabolism and activation of carcinogen. , 1998, Environmental health perspectives.

[36]  J. Thomson,et al.  Epigenetic profiles as defined signatures of xenobiotic exposure. , 2014, Mutation research. Genetic toxicology and environmental mutagenesis.

[37]  M. Hauschild,et al.  USEtox human exposure and toxicity factors for comparative assessment of toxic emissions in life cycle analysis: sensitivity to key chemical properties , 2011 .

[38]  N. Patel,et al.  Applications of linking PBPK and PD models to predict the impact of genotypic variability, formulation differences, differences in target binding capacity and target site drug concentrations on drug responses and variability , 2014, Front. Pharmacol..

[39]  Jonathan I Levy,et al.  Science and Decisions: Advancing Risk Assessment , 2010, Risk analysis : an official publication of the Society for Risk Analysis.

[40]  J. T. Macgregor,et al.  IWGT report on quantitative approaches to genotoxicity risk assessment II. Use of point-of-departure (PoD) metrics in defining acceptable exposure limits and assessing human risk. , 2015, Mutation research. Genetic toxicology and environmental mutagenesis.

[41]  David M. Reif,et al.  High-throughput models for exposure-based chemical prioritization in the ExpoCast project. , 2013, Environmental science & technology.

[42]  M E Meek,et al.  New developments in the evolution and application of the WHO/IPCS framework on mode of action/species concordance analysis , 2013, Journal of applied toxicology : JAT.

[43]  Daniel L Villeneuve,et al.  Adverse outcome pathways: A conceptual framework to support ecotoxicology research and risk assessment , 2010, Environmental toxicology and chemistry.

[44]  Mark A. J. Huijbregts,et al.  USEtox—the UNEP-SETAC toxicity model: recommended characterisation factors for human toxicity and freshwater ecotoxicity in life cycle impact assessment , 2008 .

[45]  Joseph K. Pickrell,et al.  A Systematic Survey of Loss-of-Function Variants in Human Protein-Coding Genes , 2012, Science.

[46]  Michael D Waters,et al.  Approaches for identifying germ cell mutagens: Report of the 2013 IWGT workshop on germ cell assays(☆). , 2015, Mutation research. Genetic toxicology and environmental mutagenesis.

[47]  Steven K. Gibb Toxicity testing in the 21st century: a vision and a strategy. , 2008, Reproductive toxicology.

[48]  M. Cimino,et al.  Considerations in the U.S. Environmental Protection Agency's testing approach for mutagenicity. , 1991, Mutation research.

[49]  Joel Tickner,et al.  The Globally Harmonized System for Classification and Labelling of Chemicals , 2008 .

[50]  Alan R. Boobis,et al.  The use of mode of action information in risk assessment: Quantitative key events/dose-response framework for modeling the dose-response for key events , 2014, Critical reviews in toxicology.

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

[52]  P. Stolpman,et al.  Environmental Protection Agency , 2020, The Grants Register 2022.

[53]  J. T. Macgregor,et al.  IWGT report on quantitative approaches to genotoxicity risk assessment I. Methods and metrics for defining exposure-response relationships and points of departure (PoDs). , 2015, Mutation research. Genetic toxicology and environmental mutagenesis.

[54]  Paul White,et al.  Opportunities to integrate new approaches in genetic toxicology: An ILSI‐HESI workshop report , 2015, Environmental and molecular mutagenesis.

[55]  G. Peng,et al.  Non-coding RNAs: an emerging player in DNA damage response. , 2015, Mutation research. Reviews in mutation research.

[56]  R. Elespuru Assessment of heritable genetic effects using new genetic tools and sentinels in an era of personalized medicine , 2011, Environmental and molecular mutagenesis.

[57]  A. Zhavoronkov,et al.  The role of DNA damage and repair in aging through the prism of Koch-like criteria , 2013, Ageing Research Reviews.

[58]  Kathryn S. Porter,et al.  National Health and Nutrition Examination Survey (NHANES) , 2015 .

[59]  Division on Earth Risk Assessment in the Federal Government: Managing the Process , 1983 .

[60]  M. LeBaron,et al.  Epigenetics and chemical safety assessment. , 2010, Mutation research.

[61]  Ord,et al.  Guidance for Applying Quantitative Data to Develop Data-Derived Extrapolation Factors for Interspecies and Intraspecies Extrapolation , 2014 .

[62]  Jon A Arnot,et al.  Screening level risk assessment model for chemical fate and effects in the environment. , 2006, Environmental science & technology.

[63]  Sharon Munn,et al.  Adverse outcome pathway (AOP) development I: strategies and principles. , 2014, Toxicological sciences : an official journal of the Society of Toxicology.

[64]  Ord,et al.  Recommended Use of Body Weight 3/4 as the Default Method in Derivation of the Oral Reference Dose , 2013 .

[65]  S. Bonassi,et al.  DNA damage in non-communicable diseases: A clinical and epidemiological perspective. , 2015, Mutation research.

[66]  Ord,et al.  Guidelines for Exposure Assessment , 2014 .

[67]  Angelo Moretto,et al.  Risk assessment in the 21st century: Roadmap and matrix , 2014, Critical reviews in toxicology.

[68]  Huixiao Hong,et al.  Predicting hepatotoxicity using ToxCast in vitro bioactivity and chemical structure. , 2015, Chemical research in toxicology.

[69]  C. Allis,et al.  Covalent histone modifications — miswritten, misinterpreted and mis-erased in human cancers , 2010, Nature Reviews Cancer.