The in vivo developmental toxicity of diethylstilbestrol (DES) in rat evaluated by an alternative testing strategy

Abstract

In the present study, we evaluated an alternative testing strategy to quantitatively predict the in vivo developmental toxicity of the synthetic hormone diethylstilbestrol (DES). To this end, a physiologically based kinetic (PBK) model was defined that was subsequently used to translate concentration-response data for the in vitro developmental toxicity of DES, obtained in the ES-D3 cell differentiation assay, into predicted in vivo dose-response data for developmental toxicity. The previous studies showed that the PBK model-facilitated reverse dosimetry approach is a useful approach to quantitatively predict the developmental toxicity of several developmental toxins. The results obtained in the present study show that the PBK model adequately predicted DES blood concentrations in rats. Further studies revealed that DES tested positive in the ES-D3 differentiation assay and that DES-induced inhibition of the ES-D3 cell differentiation could be counteracted by the estrogen receptor alpha (ERα) antagonist fulvestrant, indicating that the in vitro ES-D3 cell differentiation assay was able to mimic the role of ERα reported in the mode of action underlying the developmental toxicity of DES in vivo. In spite of this, combining these in vitro data with the PBK model did not adequately predict the in vivo developmental toxicity of DES in a quantitative way. It is concluded that although the EST qualifies DES as a developmental toxin and detects the role of ERα in this process, the ES-D3 cell differentiation assay of the EST apparently does not adequately capture the processes underlying DES-induced developmental toxicity in vivo. Keywords Diethylstilbestrol • Physiologically based kinetic modelling • Reverse dosimetry • Developmental toxicity • Estrogen receptor alpha (ERα) Aziza Hussein Bakheit Adam and Mengying Zhang have contributed equally to this work.

The role of metabolism in the developmental toxicity of polycyclic aromatic hydrocarbon‐containing extracts of petroleum substances

“…With these differences in molecular size and structure, DBA is able to bind to the AhR with a higher affinity than BaP, as measured in the AhR CALUX assay (Appendix B; see Supporting Information). This may also imply that DBA displays a higher intrinsic potential for interaction with other receptors present in the ES‐D3 cells, including the retinoic acid receptor and the estrogen receptor (ER) that are known to be potentially relevant in PDT (Adam et al, ; Dimopoulou et al, ; Louisse et al, ). Hence, future studies may consider the influence of structural features and size of PAHs, particularly those present in PS extracts, on their ability to activate receptors relevant for their embryotoxicity potency.…”

“…With these differences in molecular size and structure, DBA is able to bind to the AhR with a higher affinity than BaP, as measured in the AhR CALUX assay (Appendix B; see Supporting Information). This may also imply that DBA displays a higher intrinsic potential for interaction with other receptors present in the ES-D3 cells, including the retinoic acid receptor and the estrogen receptor (ER) that are known to be potentially relevant in PDT (Adam et al, 2019;Dimopoulou et al, 2018;Louisse et al, 2011). Hence, future studies may consider the influence of structural features and size of For bioactivation of the PAHs and PAH-containing PS, the present study made use of hamster liver microsomes as it has been shown that they are more effective for bioactivation of these groups of substances in comparison with rat or mouse liver microsomes (Blackburn et al, 1986;Haugen & Peak, 1983;Hermann, 1981;Hermann et al, 1980).…”

Stem cell-based test methods

“…Recently, Agapios Sachinidis and colleagues from the University of Cologne published a review about possibilities and limitations of stem cell-based test methods in pharmacology and toxicology (Sachinidis et al, 2019[ 18 ]). In recent years, much progress has been achieved concerning in vitro techniques of liver (Godoy et al, 2013[ 7 ]; Grinberg et al, 2014[ 10 ]; Leist et al, 2017[ 15 ]; Ghallab et al, 2016[ 6 ]), kidney (Sjögren et al, 2018[ 24 ]; Jiang et al, 2018[ 11 ]; Su et al, 2016[ 25 ]; Valente et al, 2012[ 26 ]; Lee et al, 2017[ 14 ]), neuronal (Keil et al, 2018[ 12 ]; Yang et al, 2018[ 28 ]; Colaianna et al, 2017[ 5 ]; Sisnaiske et al, 2014[ 23 ]) and developmental toxicity (Adam et al, 2019[ 2 ]; Bridges et al, 2019[ 3 ]; Abbott, 2019[ 1 ]). Particularly, in developmental and reproductive toxicity testing, large numbers of animals are needed for analysis of a single compound (Krug et al, 2013[ 13 ]).…”

Roadmap for the development of alternative test methods

“…Moreover, it should be well established at which concentration ranges a gold standard control should lead to positive and negative results. For example, acetaminophen (APAP) should lead to positive test results at concentrations of 1-2 mM, while concentrations below 0.1 mM should test negative in test methods for hepatotoxicity (Godoy et al 2013 In recent years, numerous in vitro tests have been developed, particularly for nephrotoxicity (Lee et al 2017;Jiang et al 2018;Sjögren et al 2018), neurotoxicity (Colaianna et al 2017;Sisnaike et al 2014;Reffatto et al 2018;Yang et al 2018;Keil et al 2018), developmental toxicity (Adam et al 2019;Krug et al 2013;Shinde et al 2017;Rempel et al 2015;Waldmann et al 2014) and hepatotoxicity (Godoy et al 2013(Godoy et al , 2015Grinberg et al 2014Grinberg et al , 2018Reif et al 2015;Leist et al 2017), often supported by methods of systems modeling (Hoehme et al 2010;Ghallab et al 2016;Vartak et al 2016;Jansen et al 2017). However, relatively little has been done to quantitatively evaluate how well in vitro/in silico methods resemble the in vivo situation.…”

“…In recent years, numerous in vitro tests have been developed, particularly for nephrotoxicity (Lee et al 2017 ; Jiang et al 2018 ; Sjögren et al 2018 ), neurotoxicity (Colaianna et al 2017 ; Sisnaike et al 2014 ; Reffatto et al 2018 ; Yang et al 2018 ; Keil et al 2018 ), developmental toxicity (Adam et al 2019 ; Krug et al 2013 ; Shinde et al 2017 ; Rempel et al 2015 ; Waldmann et al 2014 ) and hepatotoxicity (Godoy et al 2013 , 2015 ; Grinberg et al 2014 , 2018 ; Reif et al 2015 ; Leist et al 2017 ), often supported by methods of systems modeling (Hoehme et al 2010 ; Ghallab et al 2016 ; Vartak et al 2016 ; Jansen et al 2017 ). However, relatively little has been done to quantitatively evaluate how well in vitro/in silico methods resemble the in vivo situation.…”