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Adverse outcome pathways: opportunities, limitations and open questions

  • Regulatory Toxicology
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Abstract

Adverse outcome pathways (AOPs) are a recent toxicological construct that connects, in a formalized, transparent and quality-controlled way, mechanistic information to apical endpoints for regulatory purposes. AOP links a molecular initiating event (MIE) to the adverse outcome (AO) via key events (KE), in a way specified by key event relationships (KER). Although this approach to formalize mechanistic toxicological information only started in 2010, over 200 AOPs have already been established. At this stage, new requirements arise, such as the need for harmonization and re-assessment, for continuous updating, as well as for alerting about pitfalls, misuses and limits of applicability. In this review, the history of the AOP concept and its most prominent strengths are discussed, including the advantages of a formalized approach, the systematic collection of weight of evidence, the linkage of mechanisms to apical end points, the examination of the plausibility of epidemiological data, the identification of critical knowledge gaps and the design of mechanistic test methods. To prepare the ground for a broadened and appropriate use of AOPs, some widespread misconceptions are explained. Moreover, potential weaknesses and shortcomings of the current AOP rule set are addressed (1) to facilitate the discussion on its further evolution and (2) to better define appropriate vs. less suitable application areas. Exemplary toxicological studies are presented to discuss the linearity assumptions of AOP, the management of event modifiers and compensatory mechanisms, and whether a separation of toxicodynamics from toxicokinetics including metabolism is possible in the framework of pathway plasticity. Suggestions on how to compromise between different needs of AOP stakeholders have been added. A clear definition of open questions and limitations is provided to encourage further progress in the field.

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References

  • Ames BN, Whitfield HJ Jr (1966) Frameshift mutagenesis in Salmonella. Cold Spring Harb Symp Quant Biol 31:221–225

    Article  CAS  PubMed  Google Scholar 

  • Andersen ME, Krewski D (2009) Toxicity testing in the 21st century: bringing the vision to life. Toxicol Sci 107:324–330

    Article  CAS  PubMed  Google Scholar 

  • Andersen ME, Krewski D (2010) The vision of toxicity testing in the 21st century: moving from discussion to action. Toxicol Sci 117:17–24

    Article  CAS  PubMed  Google Scholar 

  • Ankley GT, Bennett RS, Erickson RJ et al (2010) Adverse outcome pathways: a conceptual framework to support ecotoxicology research and risk assessment. Environ Toxicol Chem 29:730–741

    Article  CAS  PubMed  Google Scholar 

  • Ball N, Cronin MTD, Shen J et al (2016) Toward good read-across practice (GRAP) guidance. ALTEX 33:149–166

    PubMed  PubMed Central  Google Scholar 

  • Balmer NV, Leist M (2014) Epigenetics and transcriptomics to detect adverse drug effects in model systems of human development. Basic Clin Pharmacol Toxicol 115:59–68

    Article  CAS  PubMed  Google Scholar 

  • Balmer NV, Weng M, Zimmer B et al (2012) Epigenetic changes and disturbed neural development in a human embryonic stem cell-based model relating to the fetal valproate syndrome. Hum Mol Genet 21:4104–4114

    Article  CAS  PubMed  Google Scholar 

  • Balmer NV, Klima S, Rempel E et al (2014) From transient transcriptome responses to disturbed neurodevelopment: role of histone acetylation and methylation as epigenetic switch between reversible and irreversible drug effects. Arch Toxicol 88:1451–1468

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bal-Price A, Crofton K, Leist M et al (2015a) International STakeholder NETwork for developmental neurotoxicity (ISTNET): creating a developmental neurotoxicity (DNT) testing roadmap for regulatory purposes. Arch Toxicol 89:269–287

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bal-Price A, Crofton KM, Sachana M et al (2015b) Putative adverse outcome pathways relevant to neurotoxicity. Crit Rev Toxicol 45:83–91

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bal-Price A, Lein PJ, Keil KP et al (2017) Developing and applying the adverse outcome pathway concept for understanding and predicting neurotoxicity. Neurotoxicology 59:240–255

    Article  CAS  PubMed  Google Scholar 

  • Basketter DA, Clewell H, Kimber I et al (2012) A roadmap for the development of alternative (non-animal) methods for systemic toxicity testing—t4 report. ALTEX 29:3–91

    Article  PubMed  Google Scholar 

  • Bataller R, Brenner DA (2005) Liver fibrosis. J Clin Investig 15:209–218

    Article  Google Scholar 

  • Baumann J, Gassmann K, Masjosthusmann S et al (2016) Comparative human and rat neurospheres reveal species differences in chemical effects on neurodevelopmental key events. Arch Toxicol 90:1415–1427

    Article  CAS  PubMed  Google Scholar 

  • Becker RA, Ankley GT, Edwards SW et al (2015) Increasing scientific confidence in adverse outcome pathways: application of tailored Bradford-Hill considerations for evaluating weight of evidence. Regul Toxicol Pharmacol 72:514–537

    Article  PubMed  Google Scholar 

  • Beliaeff B, Burgeot T (2002) Integrated biomarkers response: a useful tool for ecological risk assessment. Environ Toxicol Chem 21:1316–1322

    Article  CAS  PubMed  Google Scholar 

  • Bhattacharya S (1948) A test for mutagenicity of methylcholanthrene. Nature 162:573

    Article  CAS  PubMed  Google Scholar 

  • Blaauboer BJ, Boekelheide K, Clewell HJ et al (2012) The use of biomarkers of toxicity for integrating in vitro hazard estimates into risk assessment for humans. Altex 29:411–425

    Article  PubMed  Google Scholar 

  • Bois FY, Jamei M, Clewell HJ (2010) PBPK modelling of inter-individual variability in the pharmacokinetics of environmental chemicals. Toxicology 278:256–267

    Article  CAS  PubMed  Google Scholar 

  • Bolt HM, Foth H, Hengstler JG, Degen GH (2004) Carcinogenicity categorization of chemicals-new aspects to be considered in a European perspective. Toxicol Lett 151(1):29–41

    Article  CAS  PubMed  Google Scholar 

  • Bouhifd M, Andersen ME, Baghdikian C et al (2015) The human toxome project. ALTEX 32:112–124

    Article  PubMed  PubMed Central  Google Scholar 

  • Casey WM (2016) Advances in the development and validation of test methods in the United States. Toxicol Res 32:9–14

    Article  PubMed  PubMed Central  Google Scholar 

  • Carbonell P, Lopez O, Amberg A, Pastor M, Sanz F (2017) Hepatotoxicity prediction by systems biology modeling of disturbed metabolic pathways using gene expression data. ALTEX 34(2):219-234

    PubMed  Google Scholar 

  • Clippinger AJ, Hill E, Curren R et al (2016) Bridging the gap between regulatory acceptance and industry use of non-animal methods. ALTEX 33:453–458

    PubMed  Google Scholar 

  • Crawford SE, Hartung T, Hollert H et al (2017) Green toxicology: a strategy for sustainable chemical and material development. Environ Sci Eur 29:16

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Daneshian M, Kamp H, Hengstler J et al (2016) Highlight report: launch of a large integrated European in vitro toxicology project: EU-ToxRisk. Arch Toxicol 90:1021–1024

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Daston G, Knight DJ, Schwarz M et al (2015) SEURAT: safety evaluation ultimately replacing animal testing—recommendations for future research in the field of predictive toxicology. Arch Toxicol 89:15–23

    Article  CAS  PubMed  Google Scholar 

  • Delrue N, Sachana M, Sakuratani Y et al (2016) The adverse outcome pathway concept: a basis for developing regulatory decision-making tools. Altern Lab Anim 44:417–429

    PubMed  Google Scholar 

  • Depledge MH (1994) The rational basis for the use of biomarkers as ecotoxicological tools. In: Fossi MC, Leonzio C (ed.) Nondestructive biomarkers in vertebrates, pp. 271-295

  • Drasdo D, Hoehme S, Hengstler JG (2014) How predictive quantitative modelling of tissue organisation can inform liver disease pathogenesis. J Hepatol 61:951–956

    Article  PubMed  Google Scholar 

  • Duffield JS, Forbes SJ, Constandinou CM et al (2005) Selective depletion of macrophages reveals distinct, opposing roles during liver injury and repair. J Clin Invest 115:56–65

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Edwards SW, Tan YM, Villeneuve DL et al (2016) Adverse outcome pathways-organizing toxicological information to improve decision making. J Pharmacol Exp Ther 356(1):170–181

    Article  CAS  PubMed  Google Scholar 

  • Fasbender F, Widera A, Hengstler JG et al (2016) Natural killer cells and liver fibrosis. Front Immunol 7:19

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Fehrenbacher N, Gyrd-Hansen M, Poulsen B et al (2004) Sensitization to the lysosomal cell death pathway upon immortalization and transformation. Cancer Res 64:5301–5310

    Article  CAS  PubMed  Google Scholar 

  • Fischer FC et al (2017) Modeling exposure in the Tox21 in vitro bioassays. Chem Res Toxicol 30(5):1197–1208

    Article  CAS  PubMed  Google Scholar 

  • Foghsgaard L, Wissing D, Mauch D et al (2001) Cathepsin B acts as a dominant execution protease in tumor cell apoptosis induced by tumor necrosis factor. J Cell Biol 153:999–1010

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gantner F, Leist M, Küsters S et al (1996) T cell stimulus-induced crosstalk between lymphocytes and liver macrophages results in augmented cytokine release. Exp Cell Res 229:137–146

    Article  CAS  PubMed  Google Scholar 

  • Gassmann K, Abel J, Bothe H et al (2010) Species-specific differential AhR expression protects human neural progenitor cells against developmental neurotoxicity of PAHs. Environ Health Perspect 118:1571–1577

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gerhardt E, Kügler S, Leist M et al (2001) Cascade of caspase activation in potassium-deprived cerebellar granule neurons: targets for treatment with peptide and protein inhibitors of apoptosis. Mol Cell Neurosci 17:717–731 (PubMed PMID: 11312607)

    Article  CAS  PubMed  Google Scholar 

  • Ghallab A, Cellière G, Henkel SG et al (2016) Model-guided identification of a therapeutic strategy to reduce hyperammonemia in liver diseases. J Hepatol 64:860–871

    Article  CAS  PubMed  Google Scholar 

  • Godoy P, Hewitt NJ, Albrecht U et al (2013) Recent advances in 2D and 3D in vitro systems using primary hepatocytes, alternative hepatocyte sources and non-parenchymal liver cells and their use in investigating mechanisms of hepatotoxicity, cell signaling and ADME. Arch Toxicol 87:1315–1530

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Grinberg M, Stöber RM, Edlund K et al (2014) Toxicogenomics directory of chemically exposed human hepatocytes. Arch Toxicol 88(12):2261–2287

    Article  CAS  PubMed  Google Scholar 

  • Guidance document on developing and assessing adverse outcome pathways (2017) Environment directorate joint meeting of the chemicals committee and the working party on chemicals, pesticides and biotechnology; Series on Testing and Assessment No. 184. http://www.oecd.org/officialdocuments/publicdisplaydocumentpdf/?cote=env/jm/mono(2013)6&doclanguage=en. Accessed 4 Sept 2017

  • Hamon J, Jennings P, Bois FY (2014) Systems biology modeling of omics data: effect of cyclosporine a on the Nrf2 pathway in human renal cells. BMC Syst Biol 8:76

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Hansson O, Castilho RF, Kaminski Schierle GS et al (2000) Additive effects of caspase inhibitor and lazaroid on the survival of transplanted rat and human embryonic dopamine neurons. Exp Neurol 164:102–111

    Article  CAS  PubMed  Google Scholar 

  • Hansson O, Nylandsted J, Castilho RF et al (2003) Overexpression of heat shock protein 70 in R6/2 Huntington’s disease mice has only modest effects on disease progression. Brain Res 970:47–57

    Article  CAS  PubMed  Google Scholar 

  • Hartung T (2009) Food for thought… on evidence-based toxicology. ALTEX 26:75–82

    Article  PubMed  Google Scholar 

  • Hartung T (2013) Look back in anger—what clinical studies tell us about preclinical work. ALTEX 30:275–291

    Article  PubMed  PubMed Central  Google Scholar 

  • Hartung T (2016) Making big sense from big data in toxicology by read-across. ALTEX 33:83–93

    Article  PubMed  Google Scholar 

  • Hartung T (2017) Utility of the adverse outcome pathway concept in drug development. Expert Opin Drug Metab Toxicol 13:1–3

    Article  PubMed  Google Scholar 

  • Hartung T, McBride M (2011) Food for thought… on mapping the human toxome. ALTEX 28:83–93

    Article  PubMed  Google Scholar 

  • Hartung T, van Vliet E, Jaworska J et al (2012) Systems toxicology. ALTEX 29:119–128

    Article  PubMed  Google Scholar 

  • Hartung T, Luechtefeld T, Maertens A et al (2013a) Integrated testing strategies for safety assessments. ALTEX 30:3–18

    Article  PubMed  PubMed Central  Google Scholar 

  • Hartung T, Stephens M, Hoffmann S (2013b) Mechanistic validation. ALTEX 30:119–130

    Article  PubMed  PubMed Central  Google Scholar 

  • Hartung T, FitzGerald R, Paul J et al (2017) Systems toxicology—real world applications and opportunities. Chem Res Toxicol 30:870–882

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hengstler JG, Arand M, Herrero ME et al (1998) Polymorphisms of N-acetyltransferases, glutathione S-transferases, microsomal epoxide hydrolase and sulfotransferases: influence on cancer susceptibility. Recent Results Cancer Res 154:47–85

    Article  CAS  PubMed  Google Scholar 

  • Hengstler JG, Van der Burg B, Steinberg P et al (1999) Interspecies differences in cancer susceptibility and toxicity. Drug Metab Rev 31:917–970

    Article  CAS  PubMed  Google Scholar 

  • Hengstler JG, Bogdanffy MS, Bolt HM et al (2003) Challenging dogma: thresholds for genotoxic carcinogens? The case of vinyl acetate. Annu Rev Pharmacol Toxicol 43:485–520

    Article  CAS  PubMed  Google Scholar 

  • Hengstler JG, Marchan R, Leist M (2012) Highlight report: towards the replacement of in vivo repeated dose systemic toxicity testing. Arch Toxicol 86:13–15

    Article  CAS  PubMed  Google Scholar 

  • Hirt U, Gantner F, Leist M (2000) Phagocytosis of non-apoptotic cells dying by caspase-independent mechanisms. J Immunol 164:6520–6529

    Article  CAS  PubMed  Google Scholar 

  • Hoehme S, Hengstler JG, Brulport M et al (2007) Mathematical modelling of liver regeneration after intoxication with CCl(4). Chem Biol Interact 168:74–93

    Article  CAS  Google Scholar 

  • Hoehme S, Brulport M, Bauer A et al (2010) Prediction and validation of cell alignment along microvessels as order principle to restore tissue architecture in liver regeneration. Proc Natl Acad Sci USA 107:10371–10376

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hoffmann S, Hartung T (2006) Towards an evidence-based toxicology. Hum Exp Toxicol 25:497–513

    Article  CAS  PubMed  Google Scholar 

  • Hoffmann S, de Vries RBM, Stephens ML et al (2017) A primer on systematic reviews in toxicology. Arch Toxicol 91(7):2551–2575

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Horvat T, Landesmann B, Lostia A et al (2017) Adverse outcome pathway development from protein alkylation to liver fibrosis. Arch Toxicol 91:1523–1543

    Article  CAS  PubMed  Google Scholar 

  • Huggett RJ, Kimerle RA, Mehrle PM, Bergman HL (1992) Biomarkers. Biochemical, physiological, and histological markers of anthropogenic stress. Lewis Publishers, Boca Raton, p 347

    Google Scholar 

  • Jacobs MN, Colacci A, Louekari K et al (2016) International regulatory needs for development of an IATA for non-genotoxic carcinogenic chemical substances. ALTEX 33:359–392

    PubMed  Google Scholar 

  • Jalan R, Moreau R, Kamath PS, Arroyo V (2016) Acute-on-chronic liver failure: a distinct clinical condition. Semin Liver Dis 36(2):107–108

    Article  PubMed  Google Scholar 

  • Jansen PL, Ghallab A, Vartak N et al (2017) The ascending pathophysiology of cholestatic liver disease. Hepatology 65(2):722–738

    Article  CAS  PubMed  Google Scholar 

  • Jennings P (2013) Stress response pathways, toxicity pathways and adverse outcome pathways. Arch Toxicol 87:13–14

    Article  CAS  PubMed  Google Scholar 

  • Jennings P, Limonciel A, Felice L et al (2013) An overview of transcriptional regulation in response to toxicological insult. Arch Toxicol 87:49–72

    Article  CAS  PubMed  Google Scholar 

  • Kim KH, Chen CC, Monzon RI et al (2013) Matricellular protein CCN1 promotes regression of liver fibrosis through induction of cellular senescence in hepatic myofibroblasts. Mol Cell Biol 33:2078–2090

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kisseleva T, Cong M, Paik Y et al (2012) Myofibroblasts revert to an inactive phenotype during regression of liver fibrosis. Proc Natl Acad Sci USA 109:9448–9453

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kleensang A, Maertens A, Rosenberg M et al (2014) Pathways of toxicity. ALTEX 31:53–61

    Article  PubMed  Google Scholar 

  • Kleinstreuer NC, Ceger PC, Allen DG et al (2016) A curated database of rodent uterotrophic bioactivity. Environ Health Perspect 124:556–562

    PubMed  Google Scholar 

  • Kleinstreuer NC, Ceger P, Watt ED et al (2017) Development and validation of a computational model for androgen receptor activity. Chem Res Toxicol 30:946–964

    Article  CAS  PubMed  Google Scholar 

  • Knapen D, Vergauwen L, Villeneuve DL et al (2015) The potential of AOP networks for reproductive and developmental toxicity assay development. Reprod Toxicol 56:52–55

    Article  CAS  PubMed  Google Scholar 

  • Krizhanovsky V, Yon M, Dickins RA et al (2008) Senescence of activated stellate cells limits liver fibrosis. Cell 134:657–667

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Krug AK, Balmer NV, Matt F et al (2013a) Evaluation of a human neurite growth assay as specific screen for developmental neurotoxicants. Arch Toxicol 87:2215–2231

    Article  CAS  PubMed  Google Scholar 

  • Krug AK, Kolde R, Gaspar JA et al (2013b) Human embryonic stem cell-derived test systems for developmental neurotoxicity: a transcriptomics approach. Arch Toxicol 87:123–143

    Article  CAS  PubMed  Google Scholar 

  • Krug AK, Gutbier S, Zhao L et al (2014) Transcriptional and metabolic adaptation of human neurons to the mitochondrial toxicant MPP(+). Cell Death Dis. 5:e1222

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kuegler PB, Zimmer B, Waldmann T et al (2010) Markers of murine embryonic and neural stem cells, neurons and astrocytes: reference points for developmental neurotoxicitiy testing—a review by the Transatlantic Think Tank for toxicology (t4). ALTEX 27:17–42

    PubMed  Google Scholar 

  • Latta M, Künstle G, Leist M et al (2000) Metabolic depletion of ATP by fructose inversely controls CD95- and tumor necrosis factor receptor 1-mediated hepatic apoptosis. J Exp Med 191:1975–1985

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Leist M, Hartung T (2013) Inflammatory findings on species extrapolations: humans are definitely no 70-kg mice. Arch Toxicol 87:563–567

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Leist M, Jäättelä M (2001) Four deaths and a funeral: from caspases to alternative mechanisms. Nat Rev Mol Cell Biol 2:589–598

    Article  CAS  PubMed  Google Scholar 

  • Leist M, Nicotera P (1997) Calcium and neuronal death. Rev Physiol Biochem Pharmacol 132:79–125

    Article  Google Scholar 

  • Leist M, Gantner F, Künstle G et al (1996) The 55-kD tumor necrosis factor receptor and CD95 independently signal murine hepatocyte apoptosis and subsequent liver failure. Mol Med 2:109–124

    CAS  PubMed  PubMed Central  Google Scholar 

  • Leist M, Fava E, Montecucco C, Nicotera P (1997a) Peroxynitrite and nitric oxide donors induce neuronal apoptosis by eliciting autocrine excitotoxicity. Eur J Neurosci 9:1488–1498

    Article  CAS  PubMed  Google Scholar 

  • Leist M, Gantner F, Naumann H et al (1997b) Tumor necrosis factor-induced apoptosis during the poisoning of mice with hepatotoxins. Gastroenterology 112:923–934

    Article  CAS  PubMed  Google Scholar 

  • Leist M, Volbracht C, Kühnle S et al (1997c) Caspase-mediated apoptosis in neuronal excitotoxicity triggered by nitric oxide. Mol Med 3:750–764

    CAS  PubMed  PubMed Central  Google Scholar 

  • Leist M, Gantner F, Künstle G et al (1998a) Cytokine-mediated hepatic apoptosis. Rev Physiol Biochem Pharmacol 133:109–155

    CAS  PubMed  Google Scholar 

  • Leist M, Volbracht C, Fava E et al (1998b) 1-Methyl-4-phenylpyridinium induces autocrine excitotoxicity, protease activation, and neuronal apoptosis. Mol Pharmacol 54:789–801

    CAS  PubMed  Google Scholar 

  • Leist M, Hartung T, Nicotera P (2008) The dawning of a new age of toxicology. ALTEX 25:103–114

    Article  PubMed  Google Scholar 

  • Leist M, Efremova L, Karreman C (2010) Food for thought on considerations and guidelines for basic test method descriptions in toxicology. ALTEX 27:309–317

    Article  PubMed  Google Scholar 

  • Leist M, Hasiwa N, Daneshian M et al (2012) Validation and quality control of replacement alternatives—current status and future challenges. Toxicol Res 1:8–22

    Article  CAS  Google Scholar 

  • Leist M, Hasiwa N, Rovida C et al (2014) Consensus report on the future of animal-free systemic toxicity testing. ALTEX 31:341–356

    PubMed  Google Scholar 

  • Lewin L (1885) Lehrbuch der Toxikologie: für Aerzte, Studierende und Apotheker (translation: textbook of Toxicology for physicians, students and pharmacists). Urban & Schwarzenberg, Wien

    Google Scholar 

  • Lewin L (1924) Phantastica. Die betäubenden und erregenden Genussmittel. Für Ärzte und Nichtärzte (translation: Phantastica. Anaesthetic and stimulating drugs for physicians and laymen). Verlag von Georg Stilke, Berlin

  • Limonciel A, Aschauer L, Wilmes A et al (2011) Lactate is an ideal non-invasive marker for evaluating temporal alterations in cell stress and toxicity in repeat dose testing regimes. Toxicol In Vitro 25:1855–1862

    Article  CAS  PubMed  Google Scholar 

  • Maertens A, Anastas N, Spencer PJ et al (2014) Green Toxicology. ALTEX 31:243–249

    Article  PubMed  Google Scholar 

  • Matozzo V, Gagné F, Marin MG, Ricciardi F, Blaise C (2008) Vitellogenin as a biomarker of exposure to estrogenic compounds in aquatic invertebrates: a review. Environ Int 34:531–545

    Article  CAS  PubMed  Google Scholar 

  • McCarty JF, Shugart LR (1990) Biomarkers of environmental contamination. Lewis Publishers, CRC Press, Boca Raton, p 457

    Google Scholar 

  • Nicotera P, Leist M, Manzo L (1999) Neuronal cell death: a demise with different shapes. Trends Pharmacol Sci 20(2):46–51

    Article  CAS  PubMed  Google Scholar 

  • NRC (2007) Toxicity testing in the 21st century: a vision and a strategy. The National Academies Press, Washington

    Google Scholar 

  • NTP (2017) https://ntp.niehs.nih.gov/pubhealth/evalatm/test-method-evaluations/immunotoxicity/nonanimal/index.html. Accessed 4 Sept 2017

  • Nyffeler J, Karreman C, Leisner H et al (2017) Design of a high-throughput human neural crest cell migration assay to indicate potential developmental toxicants. ALTEX 34:7555–7594

    Google Scholar 

  • Nylandsted J, Wick W, Hirt UA et al (2002) Eradication of glioblastoma, and breast and colon carcinoma xenografts by Hsp70 depletion. Cancer Res 62:7139–7142

    CAS  PubMed  Google Scholar 

  • Obiol-Pardo C, Gomis-Tena J, Sanz F, Saiz J, Pastor M (2011) A multiscale simulation system for the prediction of drug-induced cardiotoxicity. J Chem Inf Model 51:483–492

    Article  CAS  PubMed  Google Scholar 

  • Ockleford C et al (2017) EFSA panel on plant protection products and their residues (PPR) investigation into experimental toxicological properties of plant protection products having a potential link to Parkinson’s disease and childhood leukaemia. EFSA J 15:4691

    Google Scholar 

  • Oesch F, Herrero ME, Hengstler JG, Lohmann M, Arand M (2000) Metabolic detoxification: implications for thresholds. Toxicol Pathol 28(3):382–387

    Article  CAS  PubMed  Google Scholar 

  • Oesch F, Herrero ME, Lohmann M, Hengstler JG, Arand M (2001) Sequestration of biological reactive intermediates by trapping as covalent enzyme-intermediate complex. Adv Exp Med Biol 500:577–586

    Article  CAS  PubMed  Google Scholar 

  • Oki NO, Edwards SW (2016) An integrative data mining approach to identifying adverse outcome pathway signatures. Toxicology 28(350–352):49–61

    Article  CAS  Google Scholar 

  • Paparella M, Colacci A, Jacobs MN (2017) Uncertainties of testing methods: what do we (want to) know about carcinogenicity? ALTEX 34(2):235–252

    PubMed  Google Scholar 

  • Patlewicz G, Ball N, Becker RA et al (2014) Read-across approaches—misconceptions, promises and challenges ahead. ALTEX 31:387–396

    Article  PubMed  Google Scholar 

  • Pelkonen O, Terron A, Hernandez AF et al (2017) Chemical exposure and infant leukaemia: development of an adverse outcome pathway (AOP) for aetiology and risk assessment research. Arch Toxicol 91(8):2763–2780

    Article  CAS  PubMed  Google Scholar 

  • Polasek TM, Patel F, Jensen BP, Sorich MJ, Wiese MD, Doogue MP (2013) Predicted metabolic drug clearance with increasing adult age. Br J Clin Pharmacol 75(4):1019–1028

    Article  CAS  PubMed  Google Scholar 

  • Poulin P, Theil FP (2002) Prediction of pharmacokinetics prior to in vivo studies. 1. Mechanism-based prediction of volume of distribution. J Pharm Sci 91:129–156

    Article  CAS  PubMed  Google Scholar 

  • Radaeva S, Sun R, Jaruga B et al (2006) Natural killer cells ameliorate liver fibrosis by killing activated stellate cells in NKG2D-dependent and tumor necrosis factor-related apoptosis-inducing ligand-dependent manners. Gastroenterology 130:435–452

    Article  CAS  PubMed  Google Scholar 

  • Rahnenführer J, Leist M (2015) From smoking guns to footprints: mining for critical events of toxicity pathways in transcriptome data. Arch Toxicol 89:813–817

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Ramachandran P, Iredale JP (2012) Macrophages: central regulators of hepatic fibrogenesis and fibrosis resolution. J Hepatol 56:1417–1419

    Article  PubMed  Google Scholar 

  • Ramachandran P, Pellicoro A, Vernon MA et al (2012) Differential Ly-6C expression identifies the recruited macrophage phenotype, which orchestrates the regression of murine liver fibrosis. Proc Natl Acad Sci USA 109:E3186–E3195

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rempel E, Hoelting L, Waldmann T et al (2015) A transcriptome-based classifier to identify developmental toxicants by stem cell testing: design, validation and optimization for histone deacetylase inhibitors. Arch Toxicol 89:1599–1618

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rostami-Hodjegan A (2012) Physiologically based pharmacokinetics joined with in vitro–in vivo extrapolation of ADME: a marriage under the arch of systems pharmacology. Clin Pharmacol Ther 92(1):50–61

    Article  CAS  PubMed  Google Scholar 

  • Rovida C, Alépée N, Api AM et al (2015) Integrated testing strategies (ITS) for safety assessment. ALTEX 32:171–181

    Article  PubMed  Google Scholar 

  • Samuel GO, Hoffmann S, Wright R et al (2016) Guidance on assessing the methodological and reporting quality of toxicologically relevant studies: a scoping review. Environ Int 92–93:630–646

    Article  PubMed  CAS  Google Scholar 

  • Sauer JM, Hartung T, Leist M et al (2015) Systems toxicology: the future of risk assessment. Int J Toxicol 34:346–348

    Article  PubMed  PubMed Central  Google Scholar 

  • Schierle GS, Hansson O, Leist M et al (1999) Caspase inhibition reduces apoptosis and increases survival of nigral transplants. Nat Med 5:97–100

    Article  CAS  PubMed  Google Scholar 

  • Schildknecht S, Pöltl D, Nagel DM et al (2009) Requirement of a dopaminergic neuronal phenotype for toxicity of low concentrations of 1-methyl-4-phenylpyridinium to human cells. Toxicol Appl Pharmacol 241:23–35

    Article  CAS  PubMed  Google Scholar 

  • Schildknecht S, Pape R, Müller N et al (2011) Neuroprotection by minocycline caused by direct and specific scavenging of peroxynitrite. J Biol Chem 286:4991–5002

    Article  CAS  PubMed  Google Scholar 

  • Schildknecht S, Karreman C, Pöltl D et al (2013) Generation of genetically-modified human differentiated cells for toxicological tests and the study of neurodegenerative diseases. ALTEX 30:427–444

    Article  PubMed  Google Scholar 

  • Schildknecht S, Pape R, Meiser J et al (2015) Preferential extracellular generation of the active parkinsonian toxicant MPP+ by transporter-independent export of the intermediate MPDP+. Antioxid Redox Signal 23:1001–1016

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Schildknecht S, Di Monte DA, Pape R et al (2017) Tipping points and endogenous determinants of nigrostriatal degeneration by MPTP. Trends Pharmacol Sci 38:541–555

    Article  CAS  PubMed  Google Scholar 

  • Schliess F, Hoehme S, Henkel SG et al (2014) Integrated metabolic spatial-temporal model for the prediction of ammonia detoxification during liver damage and regeneration. Hepatology 60:2040–2051

    Article  CAS  PubMed  Google Scholar 

  • Shinde V, Hoelting L, Perumal SS et al (2016) Definition of transcriptome-based indices for quantitative characterization of chemically disturbed stem cell development—introduction of the STOP-Toxukn and STOP-Toxukk t. Arch Toxicol 91:839–864

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Smirnova L, Hogberg HT, Leist M et al (2014) Developmental neurotoxicity—challenges in the 21st century and in vitro opportunities. ALTEX 31:129–156

    PubMed  PubMed Central  Google Scholar 

  • Stephens ML, Betts K, Beck NB et al (2016) The emergence of systematic review in toxicology. Toxicol Sci 152:10–16

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Stiegler N, Krug A, Matt F et al (2011) Assessment of chemical-induced impairment of human neurite outgrowth by multiparametric live cell imaging in high-density cultures. Toxicol Sci 121:73–87

    Article  CAS  PubMed  Google Scholar 

  • Tacke F, Zimmermann HW (2014) Macrophage heterogeneity in liver injury and fibrosis. J Hepatol 60:1090–1096

    Article  CAS  PubMed  Google Scholar 

  • Tian Z, Chen Y, Gao B (2013) Natural killer cells in liver disease. Hepatology 57:1654–1662. doi:10.1002/hep.26115

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tollefsen KE, Scholz S, Cronin MT et al (2014) Applying adverse outcome pathways (AOPs) to support integrated approaches to testing and assessment (IATA). Reg Toxicol Pharmacol 70:629–640

    Article  Google Scholar 

  • Trevan JW (1927) The error of determination of toxicity. Proc R Soc Lond B Biol Sci 101(712):483–514

  • Tsaioun K, Blaauboer BJ, Hartung T (2016) Evidence-based absorption, distribution, metabolism, excretion (ADME) and its interplay with alternative toxicity methods. ALTEX 33:343–358

    Article  PubMed  Google Scholar 

  • Users’ handbook supplement to the guidance document for developing and assessing AOPs (2017) http://www.oecd-ilibrary.org/environment/users-handbook-supplement-to-the-guidance-document-for-developing-and-assessing-adverse-outcome-pathways_5jlv1m9d1g32-en. Accessed 4 Sept 2017

  • van der Burg B, Pieterse B, Buist H et al (2015a) A high throughput screening system for predicting chemically-induced reproductive organ deformities. Reprod Toxicol 55:95–103

    Article  PubMed  CAS  Google Scholar 

  • van der Burg B, Wedebye EB, Dietrich DR et al (2015b) The ChemScreen project to design a pragmatic alternative approach to predict reproductive toxicity of chemicals. Reprod Toxicol 55:114–123

    Article  PubMed  CAS  Google Scholar 

  • van Thriel C, Westerink R, Beste C et al (2011) Translating neurobehavioural endpoints of developmental neurotoxicity tests into in vitro assays and readouts. NeuroToxicol 33:911–924

    Article  CAS  Google Scholar 

  • Vartak N, Damle-Vartak A, Richter B et al (2016) Cholestasis-induced adaptive remodeling of interlobular bile ducts. Hepatology 63:951–964

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Villeneuve DL, Crump D, Garcia-Reyero N, Hecker M, Hutchinson TH, LaLone CA, Landesmann B, Lettieri T, Munn S, Nepelska M, Ottinger MA, Vergauwen L, Whelan M (2014a) Adverse outcome pathway (AOP) development I: strategies and principles. Toxicol Sci 142:312–320

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Villeneuve DL, Crump D, Garcia-Reyero N, Hecker M, Hutchinson TH, LaLone CA, Landesmann B, Lettieri T, Munn S, Nepelska M, Ottinger MA, Vergauwen L, Whelan M (2014b) Adverse outcome pathway development. II: best practices. Toxicol Sci 142:321–330

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Vinken M (2015) Adverse outcome pathways and drug-induced liver injury testing. Chem Res Toxicol 28:1391–1397

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Volbracht C, Leist M, Nicotera P (1999) ATP controls neuronal apoptosis triggered by microtubule breakdown or potassium deprivation. Mol Med. 5:477–489

    CAS  PubMed  PubMed Central  Google Scholar 

  • Volbracht C, Fava E, Leist M et al (2001a) Calpain inhibitors prevent nitric oxide-triggered excitotoxic apoptosis. NeuroReport 12:3645–3648

    Article  CAS  PubMed  Google Scholar 

  • Volbracht C, Leist M, Kolb SA et al (2001b) Apoptosis in caspase-inhibited neurons. Mol Med 7:36–48

    CAS  PubMed  PubMed Central  Google Scholar 

  • Wilmes A, Limonciel A, Aschauer L et al (2013) Application of integrated transcriptomic, proteomic and metabolomic profiling for the delineation of mechanisms of drug induced cell stress. J Proteomics 79:180–194

    Article  CAS  PubMed  Google Scholar 

  • Wilmes A, Bielow C, Ranninger C et al (2015) Mechanism of cisplatin proximal tubule toxicity revealed by integrating transcriptomics, proteomics, metabolomics and biokinetics. Toxicol In Vitro 30:117–127

    Article  CAS  PubMed  Google Scholar 

  • Wittwehr C, Aladjov H, Ankley G et al (2017) How adverse outcome pathways can aid the development and use of computational prediction models for regulatory toxicology. Toxicol Sci 155:326–336

    Article  PubMed  Google Scholar 

  • Wynn TA (2008) Cellular and molecular mechanisms of fibrosis. J Pathol 214:199–210

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhu H, Bouhifd M, Kleinstreuer N et al (2016) Supporting read-across using biological data. ALTEX 33:167–182

    PubMed  PubMed Central  Google Scholar 

  • Zimmer B, Lee G, Balmer NV et al (2012) Evaluation of developmental toxicants and signaling pathways in a functional test based on the migration of human neural crest cells. Environ Health Perspect 120:1116–1122

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zimmer B, Pallocca G, Dreser N et al (2014) Profiling of drugs and environmental chemicals for functional impairment of neural crest migration in a novel stem cell-based test battery. Arch Toxicol 88:1109–1126

    CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This work was supported by the EU-ToxRisk project (An Integrated European “Flagship” Program Driving Mechanism-Based Toxicity Testing and Risk Assessment for the 21st Century) funded by the European Commission under the Horizon 2020 programme (Grant Agreement No. 681002). This review is a joint activity of members of the EU-ToxRisk project and additionally incorporates ideas and suggestions of several colleagues who are not members of this consortium. We thank in particular Brigitte Landesmann, Maurice Whelan, Anna Bal-Price, and Christian Desaintes for their valuable discussion. Some work on examples was funded by the German Federal Ministry of Education and Research (BMBF) (LiSyM, SysDT, NeuriTox, LivSys, Lebersimulator projects). We thank M. Turajski and B. Schanze for valuable bibliographic support, and B. Barton for excellent support in handling the manuscript.

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Authors and Affiliations

  1. In Vitro Toxicology and Biomedicine, Department inaugurated by the Doerenkamp-Zbinden Foundation, University of Konstanz, Box 657, Universitaetsstr. 10, 78457, Konstanz, Germany

    Marcel Leist, Stefan Schildknecht & Tanja Waldmann

  2. Leibniz Research Centre for Working Environment and Human Factors, Technical University Dortmund, Dortmund, Germany

    Ahmed Ghallab, Rosemarie Marchan, Reham Hassan, Patricio Godoy, Raymond Reif, Christoph van Thriel, Cristina Cadenas & Jan G. Hengstler

  3. Department of Forensic Medicine and Toxicology, Faculty of Veterinary Medicine, South Valley University, Qena, Egypt

    Ahmed Ghallab & Reham Hassan

  4. Research Division of Drug Discovery and Safety, Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands

    Rabea Graepel, Erik Danen, Joost Beltman & Bob van de Water

  5. The Danish EPA, Copenhagen, Denmark

    Susanne Hougaard Bennekou

  6. Division of Physiology, Department of Physiology and Medical Physics, Medical University of Innsbruck, 6020, Innsbruck, Austria

    Alice Limonciel

  7. Department of In Vitro Toxicology and Dermato-Cosmetology, Vrije Universiteit Brussel, Laarbeeklaan 103, 1090, Brussels, Belgium

    Mathieu Vinken

  8. BASF SE, Experimental Toxicology and Ecology, Ludwigshafen am Rhein, Germany

    Ben van Ravenzwaay & Hennicke Kamp

  9. Simcyp (A Certara Company), Sheffield, UK

    Iain Gardner & Ciarán Fisher

  10. INERIS, DRC/VIVA/METO, Parc ALATA BP2, 60550, Verneuil en Halatte, France

    Frederic Y. Bois & Enrico Mombelli

  11. Department of Food Safety, German Federal Institute for Risk Assessment, Max-Dohrn-Str. 8-10, 10589, Berlin, Germany

    Albert Braeuning

  12. Institute of Toxicology, University of Mainz, Mainz, Germany

    Franz Oesch

  13. INRIA, Unit Rocquencourt, B.P.105, 78153, Le Chesnay Cedex, France

    Dirk Drasdo

  14. Laboratoire Jacques-Louis Lions, France Université of Paris 06, CNRS, UMR 7598, 4 pl. Jussieu, Paris, France

    Dirk Drasdo

  15. Institute for Computer Science, University of Leipzig, Haertelstraße 16-18, 04107, Leipzig, Germany

    Stefan Höhme

  16. Department of Toxicology, Institute of Experimental and Clinical Pharmacology and Toxicology, Eberhard Karls University, Tübingen, Germany

    Michael Schwarz

  17. Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, USA

    Thomas Hartung

  18. Aquatic Ecology and Toxicology Section, Centre for Organismal Studies, University of Heidelberg, Im Neuenheimer Feld 504, 69120, Heidelberg, Germany

    Thomas Braunbeck

  19. Department of Human Genetics, Leiden University Medical Center, Postal Zone S4-P, PO Box 9600, 2300 RC, Leiden, The Netherlands

    Harry Vrieling

  20. Research Programme on Biomedical Informatics (GRIB), Department of Health and Life Sciences, Hospital del Mar Medical Research Institute (IMIM), Universitat Pompeu Fabra, Barcelona, Spain

    Ferran Sanz

  21. Unit of Toxicology Sciences, Swetox, Karolinska Institutet, Forskargatan 20, SE-151 36, Södertälje, Sweden

    Anna Forsby

  22. Laboratory of Environmental Chemistry and Toxicology, Department of Environmental Health Sciences, IRCCS-Istituto di Ricerche Farmacologiche Mario Negri, Via la Masa 19, 20156, Milano, Italy

    Domenico Gadaleta

  23. InSphero AG, Wagistrasse 27, CH-8952, Schlieren, Switzerland

    Jens Kelm & David Fluri

  24. Pharmacoinformatics Research Group, Department of Pharmaceutical Chemistry, University of Vienna, Althanstraße 14, 1090, Vienna, Austria

    Gerhard Ecker & Barbara Zdrazil

  25. European Food Safety Authority (EFSA), Pesticide Unit, via Carlo Magno 1 A, 43126, Parma, Italy

    Andrea Terron

  26. Division of Molecular and Computational Toxicology, Amsterdam Institute for Molecules, Medicines and Systems, Vrije Universiteit Amsterdam, De Boelelaan 1108, 1081 HZ, Amsterdam, The Netherlands

    Paul Jennings

  27. BioDetection Systems b.v. (BDS), Science Park 406, 1098 XH, Amsterdam, The Netherlands

    Bart van der Burg

  28. Section Molekular Hepatology, II. Medical Clinic, Medical Faculty Mannheim, University of Heidelberg, Theodor-Kutzer-Ufer 1-3, 68167, Mannheim, Germany

    Steven Dooley

  29. Institute of Biology, Leiden University, 2333 CC, Leiden, The Netherlands

    Annemarie H. Meijer & Bjorn Koch

  30. Department of Bioinformatics, BiGCaT, NUTRIM, Maastricht University, PO Box 616, 6200 MD, Maastricht, The Netherlands

    Egon Willighagen, Marvin Martens & Chris Evelo

  31. Open PHACTS Foundation, Cambridge, UK

    Egon Willighagen & Chris Evelo

  32. Inserm UMR-S973, Molécules Thérapeutiques in silico, Paris Diderot University, Paris, France

    Olivier Taboureau

  33. Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark

    Olivier Taboureau

  34. Istituto Superiore di Sanità, Viale Regina Elena 299, 00161, Roma, Italy

    Alberto Mantovani

  35. Douglas Connect GmbH, Bärmeggenweg 14, Zeiningen, 4314, Aargau, Switzerland

    Barry Hardy

  36. Fraunhofer Institute of Toxicology and Experimental Medicine (ITEM) Chemical Risk Assessment Group Manager Structure Activity Relationships/databases, and expert systems Nikolai-Fuchs-Strasse 1, 30625, Hannover, Germany

    Sylvia Escher

  37. TNO (Department of Risk Analysis of Products in Development), Zeist, The Netherlands

    D. Kroese

  38. Department of Neurochemistry, Stockholm University, Svante Arrhenius väg 16, SE-106 91, Stockholm, Sweden

    Anna Forsby

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Correspondence to Marcel Leist or Jan G. Hengstler.

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Leist, M., Ghallab, A., Graepel, R. et al. Adverse outcome pathways: opportunities, limitations and open questions. Arch Toxicol 91, 3477–3505 (2017). https://doi.org/10.1007/s00204-017-2045-3

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