medlitГигиена и санитарияHygiene and Sanitation0016-99002412-0650Federal Scientific Center of Hygiene named after F.F. Erisman10.47470/0016-9900-2024-103-9-1056-1061ftutujmedlit-4370Research ArticleПРОФИЛАКТИЧЕСКАЯ ТОКСИКОЛОГИЯ И ГИГИЕНИЧЕСКОЕ НОРМИРОВАНИЕPREVENTIVE TOXICOLOGY AND HYGIENIC STANDARTIZATIONСовременные альтернативные методы исследования в генетической токсикологии (обзор литературы)Modern alternative research methods in genetic toxicology (literature review)https://orcid.org/0000-0003-4748-8771ЕгороваОльга ВалерьевнаEgorovaOlga V.

Канд. биол. наук, вед. науч. сотр. отд. генетической токсикологии ФБУН «ФНЦГ им. Ф.Ф. Эрисмана» Роспотребнадзора, 141014, Мытищи, Россия

e-mail: egorova.ov@fncg.ru

MD, PhD, leading researcher of the Dept. of Genetic Toxicology of the Institute of Hygiene, Toxicology of Pesticides and Chemical Safety of the Federal Scientific Center of Hygiene named after F.F. Erisman of the Federal Service for Supervision in Protection of the Rights of Consumer and Man Wellbeing, Mytishchi, 141014, Russian Federation

e-mail: egorova.ov@fncg.ru

egorova.ov@fncg.ruИлюшинаНаталия АлексеевнаIlyushinaNatalia A.

Доктор биол. наук, зав. отд. генетической токсикологии ФБУН «ФНЦГ им. Ф.Ф. Эрисмана» Роспотребнадзора, 141014, Мытищи, Россия

e-mail: ilushina.na@fncg.ru

MD, PhD, DSci., head of the Dept. of Genetic Toxicology of the Institute of Hygiene, Toxicology of Pesticides and Chemical Safety of the Federal Scientific Center of Hygiene named after F.F. Erisman of the Federal Service for Supervision in Protection of the Rights of Consumer and Man Wellbeing, Mytishchi, 141014, Russian Federation

e-mail: ilushina.na@fncg.ru

ilushina.na@fncg.ruФБУН «Федеральный научный центр гигиены имени Ф.Ф. Эрисмана» Федеральной службы по надзору в сфере защиты прав потребителей и благополучия человекаРоссияFederal Scientific Center of Hygiene named after F.F. Erisman of the Federal Service for Supervision in Protection of the Rights of Consumer and Man WellbeingRussian Federation202416102024103910561061Copyright © Егорова О.В., Илюшина Н.А., 20242024Егорова О.В., Илюшина Н.А.Egorova O.V., Ilyushina N.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.rjhas.ru/jour/article/view/4370

Рассмотрены общие принципы оценки генотоксичности химических веществ. Основное внимание уделено альтернативным методам исследования. Обсуждается международный опыт применения альтернативных подходов и перспективы их использования в регуляторных целях.

Материалом для настоящего обзора послужили данные отечественной и зарубежной литературы, а также интернет-ресурсов в области разработки новых альтернативных методов тестирования химических веществ на генотоксичность. При поиске информации использовали базы данных ОЭСР, Scopus, Medline, Google Scholar, РИНЦ, КиберЛенинка.

Оценка генотоксичности химических веществ представляет собой сложившуюся систему, включающую целый арсенал валидированных методов, однако в настоящее время не утрачивают актуальности исследования, направленные на совершенствование существующих тестов, создание новых технологий, в том числе на основе альтернативных подходов.

В целом можно выделить три направления развития генетической токсикологии, включающих разработку новых методов тестирования на основе полногеномного секвенирования и применения технологий редактирования генома; развитие и апробацию количественной системы оценки эффектов дополнительно к существующему качественному подходу (мутагенно / не мутагенно) и анализ различных комбинаций методов тестирования на генотоксичность с целью выявления батареи тестов, обладающих большей прогностической способностью в отношении канцерогенных эффектов.

Для применения разрабатываемых альтернативных моделей в регуляторных целях необходимо предоставление убедительных доказательств того, что получаемые данные являются хорошими предикторами реального ответа организма на воздействие токсикантов (генотоксикантов), требуются валидация методов, стандартизация и гармонизация протоколов исследования, внесение изменений в существующую нормативную базу.

Участие авторов

Участие авторов: Егорова О.В. — концепция и дизайн исследования, сбор и анализ литературных данных, написание текста; Илюшина Н.А. — концепция и дизайн исследования, написание текста. Все соавторы — утверждение окончательного варианта статьи, ответственность за целостность всех частей статьи.

Конфликт интересов

Конфликт интересов. Авторы декларируют отсутствие явных и потенциальных конфликтов интересов в связи с публикацией данной статьи.

Финансирование

Финансирование. Исследование не имело спонсорской поддержки.

Поступила

Поступила: 03.05.2024 / Поступила после доработки: 30.05.2024 / Принята к печати: 19.06.2024 / Опубликована: 16.10.2024

The review represents the current principles of assessment of chemicals genotoxicity. The main attention is paid to alternative research methods. The international experience of the application of alternative approaches and prospects of their use for regulatory purposes are discussed.

The data for this review were collected from the Russian and foreign literature, as well as Internet resources, concerning the development of the new alternative methods for testing chemicals for genotoxicity. The OECD database, Scopus, Medline, Google Scholar, RISC, CyberLeninka were used for the information retrieval.

Although the evaluation of genotoxicity of chemical substances is the well-established and based on the battery of validated methods, the studies for improving the existing tests and developing new technologies, including the alternative approaches, continue unabated up to now.

In general, three trends of development of genetic toxicology can be outlined, including creating of new methods based on the whole-genome sequencing and the application of genome editing technologies; implementation of quantitative system of effects assessment in addition to the existing qualitative approach (mutagenic/non-mutagenic) and testing of various combinations of genotoxicity evaluation methods to identify a battery of tests with a greater predictive activity regarding carcinogenic effects.

To use the developed alternative models for regulatory purposes, it is necessary to provide convincing evidence that the data obtained are good predictors of the organism’s actual response to the effects of toxicants/genotoxicants, validation of methods, standardization, and harmonization of research protocols, and changes to the existing regulatory framework are required.

Contribution

Contribution: Egorova O.V. — concept and design of the study, collection and analysis of literary data, writing the text; Ilyushina N.A. — concept and design of the study, writing the text. All authors are responsible for the integrity of all parts of the manuscript and approval of the manuscript final version

Conflict of interest

Conflict of interest. The authors declare no conflict of interest.

Acknowledgement

Acknowledgement. The study had no sponsorship.

Received

Received: May 3, 2024 / Revised: May 30, 2024 / Accepted: June 19, 2024 / Published: October 16, 2024

генотоксичностьгенныехромосомные и геномные мутацииДНК-поврежденияметоды in vivoальтернативные моделиОЭСРgenotoxicitygenechromosomal and genomic mutationsDNA damagemethods in vivoalternative modelsOECDReferencesWHO. Chapter 4. Hazard Identification and Characterization: Toxicological and Human Studies of Environmental Health Criteria 240 (EHC 240); 2020. Available at: https://who.int/docs/default-source/food-safety/publications/section4-5-genotoxicity.pdfWHO. Chapter 4. Hazard Identification and Characterization: Toxicological and Human Studies of Environmental Health Criteria 240 (EHC 240); 2020. Available at: https://who.int/docs/default-source/food-safety/publications/section4-5-genotoxicity.pdfДурнев А.Д., Жанатаев А.К. Актуальные аспекты генетической токсикологии лекарственных средств. Ведомости Научного центра экспертизы средств медицинского применения. Регуляторные исследования и экспертиза лекарственных средств. 2022; 12(1): 90–109. https://doi.org/10.30895/1991-29192022-12-1-90-109 https://elibrary.ru/lezoaoDurnev A.D., Zhanataev A.K. Relevant aspects of drug genetic toxicology. Vedomosti Nauchnogo tsentra ekspertizy sredstv meditsinskogo primeneniya. Regulyatornye issledovaniya i ekspertiza lekarstvennykh sredstv. 2022; 12(1): 90–109. https://doi.org/10.30895/1991-29192022-12-1-90-109 https://elibrary.ru/lezoao (in Russian)Waters M.D., Stack H.F., Jackson M.A. Genetic toxicology data in the evaluation of potential human environmental carcinogens. Mutat. Res. 1999; 437(1): 21–49. https://doi.org/10.1016/s1383-5742(99)00037-xWaters M.D., Stack H.F., Jackson M.A. Genetic toxicology data in the evaluation of potential human environmental carcinogens. Mutat. Res. 1999; 437(1): 21–49. https://doi.org/10.1016/s1383-5742(99)00037-xZeiger E. Identification of rodent carcinogens and noncarcinogens using genetic toxicity tests: premises, promises, and performance. Regul. Toxicol. Pharmacol. 1998; 28(2): 85–95. https://doi.org/10.1006/rtph.1998.1234Zeiger E. Identification of rodent carcinogens and noncarcinogens using genetic toxicity tests: premises, promises, and performance. Regul. Toxicol. Pharmacol. 1998; 28(2): 85–95. https://doi.org/10.1006/rtph.1998.1234Douglas G.R., Blakey D.H., Clayson D.B. International Commission for Protection against Environmental Mutagens and Carcinogens. ICPEMC working paper No. 5. Genotoxicity tests as predictors of carcinogens: an analysis. Mutat. Res. 1988; 196(1): 83–93. https://doi.org/10.1016/0165-1110(88)90029-2Douglas G.R., Blakey D.H., Clayson D.B. International Commission for Protection against Environmental Mutagens and Carcinogens. ICPEMC working paper No. 5. Genotoxicity tests as predictors of carcinogens: an analysis. Mutat. Res. 1988; 196(1): 83–93. https://doi.org/10.1016/0165-1110(88)90029-2Kang S.H., Kwon J.Y., Lee J.K., Seo Y.R. Recent advances in in vivo genotoxicity testing: prediction of carcinogenic potential using comet and micronucleus assay in animal models. J. Cancer Prev. 2013; 18(4): 277–88. https://doi.org/10.15430/jcp.2013.18.4.277Kang S.H., Kwon J.Y., Lee J.K., Seo Y.R. Recent advances in in vivo genotoxicity testing: prediction of carcinogenic potential using comet and micronucleus assay in animal models. J. Cancer Prev. 2013; 18(4): 277–88. https://doi.org/10.15430/jcp.2013.18.4.277DeMarini D.M. The role of genotoxicity in carcinogenesis. In: Baan R.A., Stewart B.W., Straif K., eds. Tumour Site Concordance and Mechanisms of Carcinogenesis. Lyon: International Agency for Research on Cancer; 2019.DeMarini D.M. The role of genotoxicity in carcinogenesis. In: Baan R.A., Stewart B.W., Straif K., eds. Tumour Site Concordance and Mechanisms of Carcinogenesis. Lyon: International Agency for Research on Cancer; 2019.Luan Y., Honma M. Genotoxicity testing and recent advances. Genome Instab. Dis. 2022; 3(1): 1–21. https://doi.org/10.1007/s42764-021-00058-7Luan Y., Honma M. Genotoxicity testing and recent advances. Genome Instab. Dis. 2022; 3(1): 1–21. https://doi.org/10.1007/s42764-021-00058-7Dearfield K.L., Gollapudi B.B., Bemis J.C., Benz R.D., Douglas G.R., Elespuru R.K., et al. Next generation testing strategy for assessment of genomic damage: A conceptual framework and considerations. Environ. Mol. Mutagen. 2017; 58(5): 264–83. https://doi.org/10.1002/em.22045Dearfield K.L., Gollapudi B.B., Bemis J.C., Benz R.D., Douglas G.R., Elespuru R.K., et al. Next generation testing strategy for assessment of genomic damage: A conceptual framework and considerations. Environ. Mol. Mutagen. 2017; 58(5): 264–83. https://doi.org/10.1002/em.22045Menz J., Götz M.E., Gündel U., Gürtler R., Herrmann K., Hessel-Pras S., et al. Genotoxicity assessment: opportunities, challenges and perspectives for quantitative evaluations of dose-response data. Arch. Toxicol. 2023; 97(9): 2303–28. https://doi.org/10.1007/s00204-023-03553-wMenz J., Götz M.E., Gündel U., Gürtler R., Herrmann K., Hessel-Pras S., et al. Genotoxicity assessment: opportunities, challenges and perspectives for quantitative evaluations of dose-response data. Arch. Toxicol. 2023; 97(9): 2303–28. https://doi.org/10.1007/s00204-023-03553-wLiu Q., Lei Z., Zhu F., Ihsan A., Wang X., Yuan Z. A novel strategy to predict carcinogenicity of antiparasitics based on a combination of DNA lesions and bacterial mutagenicity tests. Front. Public Health. 2017; 5: 288. https://doi.org/10.3389/fpubh.2017.00288Liu Q., Lei Z., Zhu F., Ihsan A., Wang X., Yuan Z. A novel strategy to predict carcinogenicity of antiparasitics based on a combination of DNA lesions and bacterial mutagenicity tests. Front. Public Health. 2017; 5: 288. https://doi.org/10.3389/fpubh.2017.00288Хамидулина Х.Х., Тарасова Е.В., Ластовецкий М.Л. Применение программного обеспечения ОЭСР QSAR Toolbox для прогнозирования мутагенного действия химических веществ. Токсикологический вестник. 2022; 30(6): 403–13. https://doi.org/10.47470/0869-7922-2022-30-6-403-413 https://elibrary.ru/lyvxpbKhamidulina Kh.Kh., Tarasova E.V., Lastovetskiy M.L. Application of OECD QSAR toolbox software for predicting the mutagenic effects of chemicals. Toksikologicheskii vestnik. 2022; 30(6): 403–13. https://doi.org/10.47470/0869-7922-2022-30-6-403-413 https://elibrary.ru/lyvxpb (in Russian)OECD. Overview of the set of OECD genetic toxicology test guidelines and updates performed in 2014–2015 in OECD series on testing and assessment. No. 238. Paris: OECD Publishing; 2017: 1–70. Available at: https://one.oecd.org/document/ENV/JM/MONO%282016%2933/en/pdfOECD. Overview of the set of OECD genetic toxicology test guidelines and updates performed in 2014–2015 in OECD series on testing and assessment. No. 238. Paris: OECD Publishing; 2017: 1–70. Available at: https://one.oecd.org/document/ENV/JM/MONO%282016%2933/en/pdfOECD Library. Test № 471: Bacterial reverse mutation test; 2020. Available at: https://oecd-ilibrary.org/environment/test-no-471-bacterial-reverse-mutation-test_9789264071247-enOECD Library. Test № 471: Bacterial reverse mutation test; 2020. Available at: https://oecd-ilibrary.org/environment/test-no-471-bacterial-reverse-mutation-test_9789264071247-enOECD Library. Test № 490: In vitro mammalian cell gene mutation tests using the thymidine kinase gene; 2015. Available at: https://oecd-ilibrary.org/environment/test-no-490-in-vitro-mammalian-cell-gene-mutation-tests-using-the-thymidine-kinase-gene_9789264242241-enOECD Library. Test № 490: In vitro mammalian cell gene mutation tests using the thymidine kinase gene; 2015. Available at: https://oecd-ilibrary.org/environment/test-no-490-in-vitro-mammalian-cell-gene-mutation-tests-using-the-thymidine-kinase-gene_9789264242241-enOECD Library. Test № 476: In vitro mammalian cell gene mutation tests using the HPRT and XPRT genes; 2016. Available at: https://oecd-ilibrary.org/environment/test-no-476-in-vitro-mammalian-cell-gene-mutation-tests-using-the-hprt-and-xprt-genes_9789264264809-enOECD Library. Test № 476: In vitro mammalian cell gene mutation tests using the HPRT and XPRT genes; 2016. Available at: https://oecd-ilibrary.org/environment/test-no-476-in-vitro-mammalian-cell-gene-mutation-tests-using-the-hprt-and-xprt-genes_9789264264809-enOECD Library. Test № 473: In vitro mammalian chromosomal aberration test; 2016. Available at: https://oecd-ilibrary.org/environment/test-no-473-in-vitro-mammalian-chromosomal-aberration-test_9789264264649-enOECD Library. Test № 473: In vitro mammalian chromosomal aberration test; 2016. Available at: https://oecd-ilibrary.org/environment/test-no-473-in-vitro-mammalian-chromosomal-aberration-test_9789264264649-enOECD Library. Test № 487: In vitro mammalian cell micronucleus test; 2023. Available at: https://oecd-ilibrary.org/environment/test-no-487-in-vitro-mammalian-cell-micronucleus-test_9789264264861-enOECD Library. Test № 487: In vitro mammalian cell micronucleus test; 2023. Available at: https://oecd-ilibrary.org/environment/test-no-487-in-vitro-mammalian-cell-micronucleus-test_9789264264861-enKirkland D., Reeve L., Gatehouse D., Vanparys P. 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. Mutat. Res. 2011; 721(1): 27–73. https://doi.org/10.1016/j.mrgentox.2010.12.015Kirkland D., Reeve L., Gatehouse D., Vanparys P. 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. Mutat. Res. 2011; 721(1): 27–73. https://doi.org/10.1016/j.mrgentox.2010.12.015European Commission (EC). Commission Regulation (EU) No. 283/2013 of 1 March 2013 setting out the data requirements for active substances, in accordance with Regulation (EC) № 1107/2009 of the European Parliament and of the Council concerning the placing of plant protection products on the market; 2013.European Commission (EC). Commission Regulation (EU) No. 283/2013 of 1 March 2013 setting out the data requirements for active substances, in accordance with Regulation (EC) № 1107/2009 of the European Parliament and of the Council concerning the placing of plant protection products on the market; 2013.ECHA. Guidance on information requirements and chemical safety assessment. Chapter R. 7a: Endpoint specific guidance; 2017.ECHA. Guidance on information requirements and chemical safety assessment. Chapter R. 7a: Endpoint specific guidance; 2017.ECHA. Guidance on the Biocidal Products Regulation, Volume III: Human health, Part A: Information requirements; 2022.ECHA. Guidance on the Biocidal Products Regulation, Volume III: Human health, Part A: Information requirements; 2022.European Food Safety Authority (EFSA). Scientific opinion on genotoxicity testing strategies applicable to food and feed safety assessment. EFSA J. 2011; 9(9): 2379. https://doi.org/10.2903/j.efsa.2011.2379European Food Safety Authority (EFSA). Scientific opinion on genotoxicity testing strategies applicable to food and feed safety assessment. EFSA J. 2011; 9(9): 2379. https://doi.org/10.2903/j.efsa.2011.2379OECD Library. Test № 488: Transgenic Rodent Somatic and Germ Cell Gene Mutation Assays; 2022. Available at: https://oecd-ilibrary.org/environment/test-no-488-transgenic-rodent-somatic-and-germ-cell-gene-mutation-assays_9789264203907-enOECD Library. Test № 488: Transgenic Rodent Somatic and Germ Cell Gene Mutation Assays; 2022. Available at: https://oecd-ilibrary.org/environment/test-no-488-transgenic-rodent-somatic-and-germ-cell-gene-mutation-assays_9789264203907-enOECD Library. Test № 475: Mammalian Bone Marrow Chromosomal Aberration Test [OECD TG 475]; 2016. Available at: https://oecd-ilibrary.org/environment/test-no-475-mammalian-bone-marrow-chromosomal-aberration-test_9789264264786-enOECD Library. Test № 475: Mammalian Bone Marrow Chromosomal Aberration Test [OECD TG 475]; 2016. Available at: https://oecd-ilibrary.org/environment/test-no-475-mammalian-bone-marrow-chromosomal-aberration-test_9789264264786-enOECD Library. Test № 489: In vivo mammalian alkaline comet assay; 2016. Available at: https://oecd-ilibrary.org/environment/test-no-489-in-vivo-mammalian-alkaline-comet-assay_9789264264885-enOECD Library. Test № 489: In vivo mammalian alkaline comet assay; 2016. Available at: https://oecd-ilibrary.org/environment/test-no-489-in-vivo-mammalian-alkaline-comet-assay_9789264264885-enOECD Library. Test № 474: Mammalian Erythrocyte Micronucleus Test; 2016. Available at: https://oecd-ilibrary.org/environment/test-no-474-mammalian-erythrocyte-micronucleus-test_9789264264762-enOECD Library. Test № 474: Mammalian Erythrocyte Micronucleus Test; 2016. Available at: https://oecd-ilibrary.org/environment/test-no-474-mammalian-erythrocyte-micronucleus-test_9789264264762-enOECD Library. Test № 470: Mammalian Erythrocyte Pig-a Gene Mutation Assay; 2022. Available at: https://oecd-ilibrary.org/environment/test-no-470-mammalian-erythrocyte-pig-a-gene-mutation-assay_4faea90e-enOECD Library. Test № 470: Mammalian Erythrocyte Pig-a Gene Mutation Assay; 2022. Available at: https://oecd-ilibrary.org/environment/test-no-470-mammalian-erythrocyte-pig-a-gene-mutation-assay_4faea90e-enRobison T.W., Heflich R.H., Manjanatha M.G., Elespuru R., Atrakchi A., Mei N., et al. Appropriate in vivo follow-up assays to an in vitro bacterial reverse mutation (Ames) test positive investigational drug candidate (active pharmaceutical ingredient), drug-related metabolite, or drug-related impurity. Mutat. Res. Genet. Toxicol. Environ. Mutagen. 2021; 868–9: 503386. https://doi.org/10.1016/j.mrgentox.2021.503386Robison T.W., Heflich R.H., Manjanatha M.G., Elespuru R., Atrakchi A., Mei N., et al. Appropriate in vivo follow-up assays to an in vitro bacterial reverse mutation (Ames) test positive investigational drug candidate (active pharmaceutical ingredient), drug-related metabolite, or drug-related impurity. Mutat. Res. Genet. Toxicol. Environ. Mutagen. 2021; 868–9: 503386. https://doi.org/10.1016/j.mrgentox.2021.503386OECD Library. Test № 478: Genetic Toxicology: Rodent Dominant Lethal Test; 1984. Genetic Toxicology: Rodent Dominant Lethal Test. Available at: https://oecd-ilibrary.org/environment/test-no-478-genetic-toxicology-rodent-dominant-lethal-test_9789264071360-enOECD Library. Test № 478: Genetic Toxicology: Rodent Dominant Lethal Test; 1984. Genetic Toxicology: Rodent Dominant Lethal Test. Available at: https://oecd-ilibrary.org/environment/test-no-478-genetic-toxicology-rodent-dominant-lethal-test_9789264071360-enOECD Library. Test № 485: Genetic toxicology, Mouse Heritable Translocation Assay; 1986. Available at: https://oecd-ilibrary.org/environment/test-no-485-genetic-toxicology-mouse-heritable-translocation-assay_9789264071506-enOECD Library. Test № 485: Genetic toxicology, Mouse Heritable Translocation Assay; 1986. Available at: https://oecd-ilibrary.org/environment/test-no-485-genetic-toxicology-mouse-heritable-translocation-assay_9789264071506-enRegulation (EC) No 1223/2009 European parliament and of the council of 30 November 2009 on cosmetic products; 2009.Regulation (EC) No 1223/2009 European parliament and of the council of 30 November 2009 on cosmetic products; 2009.WHO. Determination of equivalence for public health pesticides and pesticide products. Report of a WHO consultation. Geneva; 2016. Available at: https://apps.who.int/iris/bitstream/10665/254751/1/WHO-HTM-NTD-WHOPES-2017.1-eng.pdfWHO. Determination of equivalence for public health pesticides and pesticide products. Report of a WHO consultation. Geneva; 2016. Available at: https://apps.who.int/iris/bitstream/10665/254751/1/WHO-HTM-NTD-WHOPES-2017.1-eng.pdfManual on Development and Use of FAO and WHO Specifications for Pesticides. FAO Plant Production and Protection Paper 228. First edition – third revision. Geneva: WHO Press; 2016. Available at: http://apps.who.int/iris/bitstream/handle/10665/246192/WHO-HTM-NTD-WHOPES-2016.4-eng.pdfManual on Development and Use of FAO and WHO Specifications for Pesticides. FAO Plant Production and Protection Paper 228. First edition – third revision. Geneva: WHO Press; 2016. Available at: http://apps.who.int/iris/bitstream/handle/10665/246192/WHO-HTM-NTD-WHOPES-2016.4-eng.pdfICH M7(R2). Guideline on assessment and control of DNA reactive (mutagenic) impurities in pharmaceuticals to limit potential carcinogenic risk – Step 5. 19/07/2023/ Reference Number: EMA/CHMP/ICH/83812/2013; 2023.ICH M7(R2). Guideline on assessment and control of DNA reactive (mutagenic) impurities in pharmaceuticals to limit potential carcinogenic risk – Step 5. 19/07/2023/ Reference Number: EMA/CHMP/ICH/83812/2013; 2023.FDA. Safety Testing of Drug Metabolites; 2020. Available at: https://fda.gov/regulatory-information/search-fda-guidance-documents/safety-testing-drug-metabolitesFDA. Safety Testing of Drug Metabolites; 2020. Available at: https://fda.gov/regulatory-information/search-fda-guidance-documents/safety-testing-drug-metabolitesFDA. M3(R2). Nonclinical Safety Studies for the Conduct of Human Clinical Trials and Marketing Authorization for Pharmaceuticals; 2010. Available at: https://fda.gov/regulatory-information/search-fda-guidance-documents/m3r2-nonclinicalsafety-studies-conduct-human-clinical-trials-and-marketing-authorizationFDA. M3(R2). Nonclinical Safety Studies for the Conduct of Human Clinical Trials and Marketing Authorization for Pharmaceuticals; 2010. Available at: https://fda.gov/regulatory-information/search-fda-guidance-documents/m3r2-nonclinicalsafety-studies-conduct-human-clinical-trials-and-marketing-authorizationvan der Zalm A.J., Barroso J., Browne P., Casey W., Gordon J., Henry T.R., et al. A framework for establishing scientific confidence in new approach methodologies. Arch. Toxicol. 2022; 96(11): 2865–79. https://doi.org/10.1007/s00204-022-03365-4van der Zalm A.J., Barroso J., Browne P., Casey W., Gordon J., Henry T.R., et al. A framework for establishing scientific confidence in new approach methodologies. Arch. Toxicol. 2022; 96(11): 2865–79. https://doi.org/10.1007/s00204-022-03365-4US Environmental Protection Agency Memorandum. Directive to prioritize efforts to reduce animal testing. Washington; 2019. Available at: https://epa.gov/sites/default/files/2019-09/documents/image2019-09-09-231249.pdfUS Environmental Protection Agency Memorandum. Directive to prioritize efforts to reduce animal testing. Washington; 2019. Available at: https://epa.gov/sites/default/files/2019-09/documents/image2019-09-09-231249.pdfStucki A.O., Barton-Maclaren T.S., Bhuller Y., Henriquez J.E., Henry T.R., Hirn C., et al. Use of new approach methodologies (NAMs) to meet regulatory requirements for the assessment of industrial chemicals and pesticides for effects on human health. Front. Toxicol. 2022; 4: 964553. https://doi.org/10.3389/ftox.2022.964553Stucki A.O., Barton-Maclaren T.S., Bhuller Y., Henriquez J.E., Henry T.R., Hirn C., et al. Use of new approach methodologies (NAMs) to meet regulatory requirements for the assessment of industrial chemicals and pesticides for effects on human health. Front. Toxicol. 2022; 4: 964553. https://doi.org/10.3389/ftox.2022.964553Fortin A.V., Long A.S., Williams A., Meier M.J., Cox J., Pinsonnault C., et al. Application of a new approach methodology (NAM)-based strategy for genotoxicity assessment of data-poor compounds. Front. Toxicol. 2023; 5: 1098432. https://doi.org/10.3389/ftox.2023.1098432Fortin A.V., Long A.S., Williams A., Meier M.J., Cox J., Pinsonnault C., et al. Application of a new approach methodology (NAM)-based strategy for genotoxicity assessment of data-poor compounds. Front. Toxicol. 2023; 5: 1098432. https://doi.org/10.3389/ftox.2023.1098432ECHA. Read-across assessment framework (RAAF); 2017.ECHA. Read-across assessment framework (RAAF); 2017.ECHA. Registration dossier – N,N,4-trimethylpiperazine-1-ethylamine; 2019. Available at: https://echa.europa.eu/nl/registration-dossier/-/registered-dossier/27533/7/7/1ECHA. Registration dossier – N,N,4-trimethylpiperazine-1-ethylamine; 2019. Available at: https://echa.europa.eu/nl/registration-dossier/-/registered-dossier/27533/7/7/1Раздольский А.Н., Григорьев В.Ю., Ярков А.В., Григорьева Л.Д., Страхова Н.Н., Казаченко В.П. и др. Классификационные QSAR модели субстратной активности химических соединений по отношению к p-гликопротеину на базе спектра межатомных внутримолекулярных взаимодействий. Международный журнал прикладных и фундаментальных исследований. 2021; (6): 97–103. https://doi.org/10.17513/mjpfi.13238 https://elibrary.ru/yynxecRazdolskii A.N., Grigor’ev V.Yu., Yarkov A.V., Grigoreva L.D., Strakhova N.N., Kazachenko V.P., et al. Classification QSAR models of substrate activity of chemical relative to p-glycoprotein based on the spectrum of interatomic intramolecular interacrtion. Mezhdunarodnyi zhurnal prikladnykh i fundamental’nykh issledovanii. 2021; (6): 97–103. https://doi.org/10.17513/mjpfi.13238 https://elibrary.ru/yynxec (in Russian)Сухачев В.С., Иванов С.М., Филимонов Д.А., Поройков В.В. Альтернативные методы исследования. Компьютерная оценка острой токсичности для грызунов. Лабораторные животные для научных исследований. 2019; (4): 4. https://doi.org/10.29926/2618723X-2019-04-04 https://elibrary.ru/nysibySukhachev V.S., Ivanov S.M., Filimonov D.A., Poroikov V.V. Alternative methods of studies. Computer-aided estimation of rodents acute toxicity. Laboratornye zhivotnye dlya nauchnykh issledovanii. 2019; (4): 4. https://doi.org/10.29926/2618723X-2019-04-04 https://elibrary.ru/nysiby (in Russian)Гусева Е.А., Николаева Н.И., Филин А.С., Савостикова О.Н. Сравнительная оценка математических моделей прогнозирования острой токсичности химических веществ. Гигиена и санитария. 2022; 101(7): 816–23. https://doi.org/10.47470/0016-9900-2022-101-7-816-823 https://elibrary.ru/trwbtpGuseva E.A., Nikolaeva N.I., Filin A.S., Savostikova O.N. Comparative evaluation of mathematical models for predicting acute toxicity of chemicals. Gigiena i Sanitaria (Hygiene and Sanitation, Russian journal). 2022; 101(7): 816–23. https://doi.org/10.47470/0016-9900-2022-101-7-816-823 https://elibrary.ru/trwbtp (in Russian)Kavlock R.J., Bahadori T., Barton-Maclaren T.S., Gwinn M.R., Rasenberg M., Thomas R.S. Accelerating the pace of chemical risk assessment. Chem. Res. Toxicol. 2018; 31(5): 287–90. https://doi.org/10.1021/acs.chemrestox.7b00339Kavlock R.J., Bahadori T., Barton-Maclaren T.S., Gwinn M.R., Rasenberg M., Thomas R.S. Accelerating the pace of chemical risk assessment. Chem. Res. Toxicol. 2018; 31(5): 287–90. https://doi.org/10.1021/acs.chemrestox.7b00339Magurany K.A., Chang X., Clewell R., Coecke S., Haugabrooks E., Marty S. A pragmatic framework for the application of new approach methodologies in one health toxicological risk assessment. Toxicol. Sci. 2023; 192(2): 155–77. https://doi.org/10.1093/toxsci/kfad012Magurany K.A., Chang X., Clewell R., Coecke S., Haugabrooks E., Marty S. A pragmatic framework for the application of new approach methodologies in one health toxicological risk assessment. Toxicol. Sci. 2023; 192(2): 155–77. https://doi.org/10.1093/toxsci/kfad012Diniz R.R., Domingos T.F.S., Pinto G.R., Cabral L.M., de Pádula M., de Souza A.M.T. Use of in silico and in vitro methods as a potential new approach methodologies (NAMs) for (photo)mutagenicity and phototoxicity risk assessment of agrochemicals. Sci. Total. Environ. 2023; 904: 167320. https://doi.org/10.1016/j.scitotenv.2023.167320Diniz R.R., Domingos T.F.S., Pinto G.R., Cabral L.M., de Pádula M., de Souza A.M.T. Use of in silico and in vitro methods as a potential new approach methodologies (NAMs) for (photo)mutagenicity and phototoxicity risk assessment of agrochemicals. Sci. Total. Environ. 2023; 904: 167320. https://doi.org/10.1016/j.scitotenv.2023.167320Liu W., Xi J., Cao Y., You X., Chen R., Zhang X., et al. An adaption of human-induced hepatocytes to in vitro genetic toxicity tests. Mutagenesis. 2019; 34(2): 165–71. https://doi.org/10.1093/mutage/gey041Liu W., Xi J., Cao Y., You X., Chen R., Zhang X., et al. An adaption of human-induced hepatocytes to in vitro genetic toxicity tests. Mutagenesis. 2019; 34(2): 165–71. https://doi.org/10.1093/mutage/gey041Chapin R.E., Stedman D.B. Endless possibilities: stem cells and the vision for toxicology testing in the 21st century. Toxicol. Sci. 2009; 112(1): 17–22. https://doi.org/10.1093/toxsci/kfp202Chapin R.E., Stedman D.B. Endless possibilities: stem cells and the vision for toxicology testing in the 21st century. Toxicol. Sci. 2009; 112(1): 17–22. https://doi.org/10.1093/toxsci/kfp202Kang K.S., Trosko J.E. Stem cells in toxicology: fundamental biology and practical considerations. Toxicol. Sci. 2011; 120(Suppl. 1): S269–89. https://doi.org/10.1093/toxsci/kfq370Kang K.S., Trosko J.E. Stem cells in toxicology: fundamental biology and practical considerations. Toxicol. Sci. 2011; 120(Suppl. 1): S269–89. https://doi.org/10.1093/toxsci/kfq370Hendriks G., Derr R.S., Misovic B., Morolli B., Calléja F.M., Vrieling H. The extended ToxTracker assay discriminates between induction of DNA damage, oxidative stress, and protein misfolding. Toxicol. Sci. 2016; 150(1): 190–203. https://doi.org/10.1093/toxsci/kfv323Hendriks G., Derr R.S., Misovic B., Morolli B., Calléja F.M., Vrieling H. The extended ToxTracker assay discriminates between induction of DNA damage, oxidative stress, and protein misfolding. Toxicol. Sci. 2016; 150(1): 190–203. https://doi.org/10.1093/toxsci/kfv323Wills J.W., Halkes-Wellstead E., Summers H.D., Rees P., Johnson G.E. Empirical comparison of genotoxic potency estimations: the in vitro DNA-damage ToxTracker endpoints versus the in vivo micronucleus assay. Mutagenesis. 2021; 36(4): 311–20. https://doi.org/10.1093/mutage/geab020Wills J.W., Halkes-Wellstead E., Summers H.D., Rees P., Johnson G.E. Empirical comparison of genotoxic potency estimations: the in vitro DNA-damage ToxTracker endpoints versus the in vivo micronucleus assay. Mutagenesis. 2021; 36(4): 311–20. https://doi.org/10.1093/mutage/geab020Available at: https://toxys.com/toxtracker-suiteAvailable at: https://toxys.com/toxtracker-suiteWilde S., Dambowsky M., Hempt C., Sutter A., Queisser N. Classification of in vitro genotoxicants using a novel multiplexed biomarker assay compared to the flow cytometric micronucleus test. Environ. Mol. Mutagen. 2017; 58(9): 662–77. https://doi.org/10.1002/em.22130Wilde S., Dambowsky M., Hempt C., Sutter A., Queisser N. Classification of in vitro genotoxicants using a novel multiplexed biomarker assay compared to the flow cytometric micronucleus test. Environ. Mol. Mutagen. 2017; 58(9): 662–77. https://doi.org/10.1002/em.22130Bryce S.M., Bernacki D.T., Bemis J.C., Spellman R.A., Engel M.E., Schuler M., et al. Interlaboratory evaluation of a multiplexed high information content in vitro genotoxicity assay. Environ. Mol. Mutagen. 2017; 58(3): 146–61. https://doi.org/10.1002/em.22083Bryce S.M., Bernacki D.T., Bemis J.C., Spellman R.A., Engel M.E., Schuler M., et al. Interlaboratory evaluation of a multiplexed high information content in vitro genotoxicity assay. Environ. Mol. Mutagen. 2017; 58(3): 146–61. https://doi.org/10.1002/em.22083Dertinger S.D., Kraynak A.R., Wheeldon R.P., Bernacki D.T., Bryce S.M., Hall N., et al. Predictions of genotoxic potential, mode of action, molecular targets, and potency via a tiered Multiflow® assay data analysis strategy. Environ. Mol. Mutagen. 2019; 60(6): 513–33. https://doi.org/10.1002/em.22274Dertinger S.D., Kraynak A.R., Wheeldon R.P., Bernacki D.T., Bryce S.M., Hall N., et al. Predictions of genotoxic potential, mode of action, molecular targets, and potency via a tiered Multiflow® assay data analysis strategy. Environ. Mol. Mutagen. 2019; 60(6): 513–33. https://doi.org/10.1002/em.22274Winkelbeiner N., Wandt V.K., Ebert F., Lossow K., Bankoglu E.E., Martin M., et al. A multi-endpoint approach to base excision repair incision activity augmented by PARylation and DNA damage levels in mice: impact of sex and age. Int. J. Mol. Sci. 2020; 21(18): 6600. https://doi.org/10.3390/ijms21186600Winkelbeiner N., Wandt V.K., Ebert F., Lossow K., Bankoglu E.E., Martin M., et al. A multi-endpoint approach to base excision repair incision activity augmented by PARylation and DNA damage levels in mice: impact of sex and age. Int. J. Mol. Sci. 2020; 21(18): 6600. https://doi.org/10.3390/ijms21186600Alépée N., Bahinski A., Daneshian M., De Wever B., Fritsche E., Goldberg A., et al. State-of-the-art of 3D cultures (organs-on-a-chip) in safety testing and pathophysiology. ALTEX. 2014; 31(4): 441–77. https://doi.org/10.14573/altex.1406111Alépée N., Bahinski A., Daneshian M., De Wever B., Fritsche E., Goldberg A., et al. State-of-the-art of 3D cultures (organs-on-a-chip) in safety testing and pathophysiology. ALTEX. 2014; 31(4): 441–77. https://doi.org/10.14573/altex.1406111Shah U.K., Mallia J.O., Singh N., Chapman K.E., Doak S.H., Jenkins G.J.S. A three-dimensional in vitro HepG2 cells liver spheroid model for genotoxicity studies. Mutat. Res. Genet. Toxicol. Environ. Mutagen. 2018; 825: 51–8. https://doi.org/10.1016/j.mrgentox.2017.12.005Shah U.K., Mallia J.O., Singh N., Chapman K.E., Doak S.H., Jenkins G.J.S. A three-dimensional in vitro HepG2 cells liver spheroid model for genotoxicity studies. Mutat. Res. Genet. Toxicol. Environ. Mutagen. 2018; 825: 51–8. https://doi.org/10.1016/j.mrgentox.2017.12.005Augustyniak J., Bertero A., Coccini T., Baderna D., Buzanska L., Caloni F. Organoids are promising tools for species-specific in vitro toxicological studies. J. Appl. Toxicol. 2019; 39(12): 1610–22. https://doi.org/10.1002/jat.3815Augustyniak J., Bertero A., Coccini T., Baderna D., Buzanska L., Caloni F. Organoids are promising tools for species-specific in vitro toxicological studies. J. Appl. Toxicol. 2019; 39(12): 1610–22. https://doi.org/10.1002/jat.3815Weinhart M., Hocke A., Hippenstiel S., Kurreck J., Hedtrich S. 3D organ models – revolution in pharmacological research? Pharmacol. Res. 2019; 139: 446–51. https://doi.org/10.1016/j.phrs.2018.11.002Weinhart M., Hocke A., Hippenstiel S., Kurreck J., Hedtrich S. 3D organ models – revolution in pharmacological research? Pharmacol. Res. 2019; 139: 446–51. https://doi.org/10.1016/j.phrs.2018.11.002Chen L., Li N., Liu Y., Faquet B., Alépée N., Ding C., et al. A new 3D model for genotoxicity assessment: EpiSkin™ micronucleus assay. Mutagenesis. 2021; 36(1): 51–61. https://doi.org/10.1093/mutage/geaa003Chen L., Li N., Liu Y., Faquet B., Alépée N., Ding C., et al. A new 3D model for genotoxicity assessment: EpiSkin™ micronucleus assay. Mutagenesis. 2021; 36(1): 51–61. https://doi.org/10.1093/mutage/geaa003Reus A., Staal Y., van Triel J., van Acker F., Kuper Human F. 3D airway models to explore in vivo inhalation. Eur. Respir. J. 2011; 38 (Suppl. 55): 3089.Reus A., Staal Y., van Triel J., van Acker F., Kuper Human F. 3D airway models to explore in vivo inhalation. Eur. Respir. J. 2011; 38 (Suppl. 55): 3089.Pfuhler S., van Benthem J., Curren R., Doak S.H., Dusinska M., Hayashi M., et al. Use of in vitro 3D tissue models in genotoxicity testing: Strategic fit, validation status and way forward. Report of the working group from the 7th International Workshop on Genotoxicity Testing (IWGT). Mutat. Res. Genet. Toxicol. Environ. Mutagen. 2020; 850–851: 503135. https://doi.org/10.1016/j.mrgentox.2020.503135Pfuhler S., van Benthem J., Curren R., Doak S.H., Dusinska M., Hayashi M., et al. Use of in vitro 3D tissue models in genotoxicity testing: Strategic fit, validation status and way forward. Report of the working group from the 7th International Workshop on Genotoxicity Testing (IWGT). Mutat. Res. Genet. Toxicol. Environ. Mutagen. 2020; 850–851: 503135. https://doi.org/10.1016/j.mrgentox.2020.503135Pfuhler S., Fellows M., van Benthem J., Corvi R., Curren R., Dearfield K., et al. In vitro genotoxicity test approaches with better predictivity: summary of an IWGT workshop. Mutat. Res. 2011; 723(2): 101–7. https://doi.org/10.1016/j.mrgentox.2011.03.013Pfuhler S., Fellows M., van Benthem J., Corvi R., Curren R., Dearfield K., et al. In vitro genotoxicity test approaches with better predictivity: summary of an IWGT workshop. Mutat. Res. 2011; 723(2): 101–7. https://doi.org/10.1016/j.mrgentox.2011.03.013National Institute of Health Sciences. TK6 Mutants Consortium. National Institute of Health Sciences; 2019. Available at: http://nihs.go.jp/dgm/tk6.htmlNational Institute of Health Sciences. TK6 Mutants Consortium. National Institute of Health Sciences; 2019. Available at: http://nihs.go.jp/dgm/tk6.htmlBoysen G., Nookaew I. Current and future methodology for quantitation and site-specific mapping the location of DNA adducts. Toxics. 2022; 10(2): 45. https://doi.org/10.3390/toxics10020045Boysen G., Nookaew I. Current and future methodology for quantitation and site-specific mapping the location of DNA adducts. Toxics. 2022; 10(2): 45. https://doi.org/10.3390/toxics10020045Swenberg J.A., Lu K., Moeller B.C., Gao L., Upton P.B., Nakamura J., et al. Endogenous versus exogenous DNA adducts: their role in carcinogenesis, epidemiology, and risk assessment. Toxicol. Sci. 2011; 120(Suppl. 1): S130–45. https://doi.org/10.1093/toxsci/kfq371Swenberg J.A., Lu K., Moeller B.C., Gao L., Upton P.B., Nakamura J., et al. Endogenous versus exogenous DNA adducts: their role in carcinogenesis, epidemiology, and risk assessment. Toxicol. Sci. 2011; 120(Suppl. 1): S130–45. https://doi.org/10.1093/toxsci/kfq371Balbo S., Turesky R.J., Villalta P.W. DNA adductomics. Chem. Res. Toxicol. 2014; 27(3): 356–66. https://doi.org/10.1021/tx4004352Balbo S., Turesky R.J., Villalta P.W. DNA adductomics. Chem. Res. Toxicol. 2014; 27(3): 356–66. https://doi.org/10.1021/tx4004352Hemeryck L.Y., Rombouts C., De Paepe E., Vanhaecke L. DNA adduct profiling of in vitro colonic meat digests to map red vs. white meat genotoxicity. Food Chem. Toxicol. 2018; 115: 73–87. https://doi.org/10.1016/j.fct.2018.02.032Hemeryck L.Y., Rombouts C., De Paepe E., Vanhaecke L. DNA adduct profiling of in vitro colonic meat digests to map red vs. white meat genotoxicity. Food Chem. Toxicol. 2018; 115: 73–87. https://doi.org/10.1016/j.fct.2018.02.032Pietsch K.E., van Midwoud P.M., Villalta P.W., Sturla S.J. Quantification of acylfulvene- and illudin S-DNA adducts in cells with variable bioactivation capacities. Chem. Res. Toxicol. 2013; 26(1): 146–55. https://doi.org/10.1021/tx300430rPietsch K.E., van Midwoud P.M., Villalta P.W., Sturla S.J. Quantification of acylfulvene- and illudin S-DNA adducts in cells with variable bioactivation capacities. Chem. Res. Toxicol. 2013; 26(1): 146–55. https://doi.org/10.1021/tx300430rThomas R.S., Wesselkamper S.C., Wang N.C., Zhao Q.J., Petersen D.D., Lambert J.C., et al. Temporal concordance between apical and transcriptional points of departure for chemical risk assessment. Toxicol. Sci. 2013; 134(1): 180–94. https://doi.org/10.1093/toxsci/kft094Thomas R.S., Wesselkamper S.C., Wang N.C., Zhao Q.J., Petersen D.D., Lambert J.C., et al. Temporal concordance between apical and transcriptional points of departure for chemical risk assessment. Toxicol. Sci. 2013; 134(1): 180–94. https://doi.org/10.1093/toxsci/kft094Farmahin R., Williams A., Kuo B., Chepelev N.L., Thomas R.S., Barton-Maclaren T.S., et al. Recommended approaches in the application of toxicogenomics to derive points of departure for chemical risk assessment. Arch. Toxicol. 2017; 91(5): 2045–65. https://doi.org/10.1007/s00204-016-1886-5Farmahin R., Williams A., Kuo B., Chepelev N.L., Thomas R.S., Barton-Maclaren T.S., et al. Recommended approaches in the application of toxicogenomics to derive points of departure for chemical risk assessment. Arch. Toxicol. 2017; 91(5): 2045–65. https://doi.org/10.1007/s00204-016-1886-5Yauk C.L., Cheung C., Barton-Maclaren T.S., Boucher S., Bourdon-Lacombe J., Chauhan V., et al. Toxicogenomic applications in risk assessment at Health Canada. Curr. Opin. Toxicol. 2019; 18: 34–45. https://doi.org/10.1016/j.cotox.2019.02.005Yauk C.L., Cheung C., Barton-Maclaren T.S., Boucher S., Bourdon-Lacombe J., Chauhan V., et al. Toxicogenomic applications in risk assessment at Health Canada. Curr. Opin. Toxicol. 2019; 18: 34–45. https://doi.org/10.1016/j.cotox.2019.02.005Li H.H., Chen R., Hyduke D.R., Williams A., Frötschl R., Ellinger-Ziegelbauer H., et al. Development and validation of a high-throughput transcriptomic biomarker to address 21st century genetic toxicology needs. Proc. Natl. Acad. Sci. USA. 2017; 114(51): E10881–9. https://doi.org/10.1073/pnas.1714109114Li H.H., Chen R., Hyduke D.R., Williams A., Frötschl R., Ellinger-Ziegelbauer H., et al. Development and validation of a high-throughput transcriptomic biomarker to address 21st century genetic toxicology needs. Proc. Natl. Acad. Sci. USA. 2017; 114(51): E10881–9. https://doi.org/10.1073/pnas.1714109114Li H.H., Yauk C.L., Chen R., Hyduke D.R., Williams A., Frötschl R., et al. TGx-DDI, a transcriptomic biomarker for genotoxicity hazard assessment of pharmaceuticals and environmental chemicals. Front. Big Data. 2019; 2: 36. https://doi.org/10.3389/fdata.2019.00036Li H.H., Yauk C.L., Chen R., Hyduke D.R., Williams A., Frötschl R., et al. TGx-DDI, a transcriptomic biomarker for genotoxicity hazard assessment of pharmaceuticals and environmental chemicals. Front. Big Data. 2019; 2: 36. https://doi.org/10.3389/fdata.2019.00036Pérez L.O., González-José R., García P.P. Prediction of non-genotoxic carcinogenicity based on genetic profiles of short-term exposure assays. Toxicol. Res. 2016; 32(4): 289–300. https://doi.org/10.5487/TR.2016.32.4.289Pérez L.O., González-José R., García P.P. Prediction of non-genotoxic carcinogenicity based on genetic profiles of short-term exposure assays. Toxicol. Res. 2016; 32(4): 289–300. https://doi.org/10.5487/TR.2016.32.4.289Felter S.P., Bhat V.S., Botham P.A., Bussard D.A., Casey W., Hayes A.W., et al. Assessing chemical carcinogenicity: hazard identification, classification, and risk assessment. Insight from a Toxicology Forum state-of-the-science workshop. Crit. Rev. Toxicol. 2021; 51(8): 653–94. https://doi.org/10.1080/10408444.2021.2003295Felter S.P., Bhat V.S., Botham P.A., Bussard D.A., Casey W., Hayes A.W., et al. Assessing chemical carcinogenicity: hazard identification, classification, and risk assessment. Insight from a Toxicology Forum state-of-the-science workshop. Crit. Rev. Toxicol. 2021; 51(8): 653–94. https://doi.org/10.1080/10408444.2021.2003295Moffat I., Chepelev N., Labib S., Bourdon-Lacombe J., Kuo B., Buick J.K., et al. Comparison of toxicogenomics and traditional approaches to inform mode of action and points of departure in human health risk assessment of benzo[a]pyrene in drinking water. Crit. Rev. Toxicol. 2015; 45(1): 1–43. https://doi.org/10.3109/10408444.2014.973934Moffat I., Chepelev N., Labib S., Bourdon-Lacombe J., Kuo B., Buick J.K., et al. Comparison of toxicogenomics and traditional approaches to inform mode of action and points of departure in human health risk assessment of benzo[a]pyrene in drinking water. Crit. Rev. Toxicol. 2015; 45(1): 1–43. https://doi.org/10.3109/10408444.2014.973934Cho E., Buick J.K., Williams A., Chen R., Li H.H., Corton J.C., et al. Assessment of the performance of the TGx-DDI biomarker to detect DNA damage-inducing agents using quantitative RT-PCR in TK6 cells. Environ. Mol. Mutagen. 2019; 60(2): 122–33. https://doi.org/10.1002/em.22257Cho E., Buick J.K., Williams A., Chen R., Li H.H., Corton J.C., et al. Assessment of the performance of the TGx-DDI biomarker to detect DNA damage-inducing agents using quantitative RT-PCR in TK6 cells. Environ. Mol. Mutagen. 2019; 60(2): 122–33. https://doi.org/10.1002/em.22257Buick J.K., Williams A., Gagné R., Swartz C.D., Recio L., Ferguson S.S., et al. Flow cytometric micronucleus assay and TGx-DDI transcriptomic biomarker analysis of ten genotoxic and non-genotoxic chemicals in human HepaRG™ cells. Genes. Environ. 2020; 42: 5. https://doi.org/10.1186/s41021-019-0139-2Buick J.K., Williams A., Gagné R., Swartz C.D., Recio L., Ferguson S.S., et al. Flow cytometric micronucleus assay and TGx-DDI transcriptomic biomarker analysis of ten genotoxic and non-genotoxic chemicals in human HepaRG™ cells. Genes. Environ. 2020; 42: 5. https://doi.org/10.1186/s41021-019-0139-2Buick J.K., Williams A., Meier M.J., Swartz C.D., Recio L., Gagné R., et al. A modern genotoxicity testing paradigm: integration of the high-throughput CometChip® and the TGx-DDI transcriptomic biomarker in human HepaRG™ cell cultures. Front. Public Health. 2021; 9: 694834. https://doi.org/10.3389/fpubh.2021.694834Buick J.K., Williams A., Meier M.J., Swartz C.D., Recio L., Gagné R., et al. A modern genotoxicity testing paradigm: integration of the high-throughput CometChip® and the TGx-DDI transcriptomic biomarker in human HepaRG™ cell cultures. Front. Public Health. 2021; 9: 694834. https://doi.org/10.3389/fpubh.2021.694834Pradeep P., Patlewicz G., Pearce R., Wambaugh J., Wetmore B., Judson R. Using chemical structure information to develop predictive models for in vitro toxicokinetic parameters to inform high-throughput risk-assessment. Comput. Toxicol. 2020; 16: 10.1016/j.comtox.2020.100136. https://doi.org/10.1016/j.comtox.2020.100136Pradeep P., Patlewicz G., Pearce R., Wambaugh J., Wetmore B., Judson R. Using chemical structure information to develop predictive models for in vitro toxicokinetic parameters to inform high-throughput risk-assessment. Comput. Toxicol. 2020; 16: 10.1016/j.comtox.2020.100136. https://doi.org/10.1016/j.comtox.2020.100136Pradeep P., Friedman K.P., Judson R. Structure-based QSAR models to predict repeat dose toxicity points of departure. Comput. Toxicol. 2020; 16(November 2020): 10.1016/j.comtox.2020.100139. https://doi.org/10.1016/j.comtox.2020.100139Pradeep P., Friedman K.P., Judson R. Structure-based QSAR models to predict repeat dose toxicity points of departure. Comput. Toxicol. 2020; 16(November 2020): 10.1016/j.comtox.2020.100139. https://doi.org/10.1016/j.comtox.2020.100139Tennant R.E., Guesné S.J., Canipa S., Cayley A., Drewe W.C., Honma M., et al. Extrapolation of in vitro structural alerts for mutagenicity to the in vivo endpoint. Mutagenesis. 2019; 34(1): 111–21. https://doi.org/10.1093/mutage/gey030Tennant R.E., Guesné S.J., Canipa S., Cayley A., Drewe W.C., Honma M., et al. Extrapolation of in vitro structural alerts for mutagenicity to the in vivo endpoint. Mutagenesis. 2019; 34(1): 111–21. https://doi.org/10.1093/mutage/gey030Zang Q., Mansouri K., Williams A.J., Judson R.S., Allen D.G., Casey W.M., et al. In silico prediction of physicochemical properties of environmental chemicals using molecular fingerprints and machine learning. J. Chem. Inf. Model. 2017; 57(1): 36–49. https://doi.org/10.1021/acs.jcim.6b00625Zang Q., Mansouri K., Williams A.J., Judson R.S., Allen D.G., Casey W.M., et al. In silico prediction of physicochemical properties of environmental chemicals using molecular fingerprints and machine learning. J. Chem. Inf. Model. 2017; 57(1): 36–49. https://doi.org/10.1021/acs.jcim.6b00625Doak S.H., Andreoli C., Burgum M.J., Chaudhry Q., Bleeker E.A.J., Bossa C., et al. Current status and future challenges of genotoxicity OECD Test Guidelines for nanomaterials: a workshop report. Mutagenesis. 2023; 38(4): 183–91. https://doi.org/10.1093/mutage/gead017Doak S.H., Andreoli C., Burgum M.J., Chaudhry Q., Bleeker E.A.J., Bossa C., et al. Current status and future challenges of genotoxicity OECD Test Guidelines for nanomaterials: a workshop report. Mutagenesis. 2023; 38(4): 183–91. https://doi.org/10.1093/mutage/gead017

The authors declare that there are no conflicts of interest present.