Predictive value of cell assays for developmental toxicity and embryotoxicity of conazole fungicides.

This paper evaluates in vivo predictability of a battery of in vitro tests covering developmental toxicity and embryotoxicity of five widely used conazole fungicides. The conazoles were investigated in the embryonic stem cell test, and data were compared to in vivo embryotoxicity data. The same conazoles were evaluated on the basis of data from a battery of cell assays for endocrine activity, including assays for AR, ER, AhR, and sex hormone synthesis, and data were compared to in vivo developmental toxicity data. Overall, the ranking of the five conazole fungicides based on in vitro data were in reasonably good agreement with available in vivo effects. Ketoconazole and epoxiconazole are the most potent embryotoxic compounds, whereas prochloraz belongs to the most potent developmental toxicants. In conclusion, a rough prediction of the ranking of these conazole fungicides for in vivo toxicity data was possible by a holistic evaluation of data from a panel of cell-based assays.

[1]  V. Berezin,et al.  Prenatal prochloraz treatment significantly increases pregnancy length and reduces offspring weight but does not affect social-olfactory memory in rats. , 2013, Experimental and toxicologic pathology : official journal of the Gesellschaft fur Toxikologische Pathologie.

[2]  L. Clemmensen,et al.  Adverse effects on sexual development in rat offspring after low dose exposure to a mixture of endocrine disrupting pesticides. , 2012, Reproductive toxicology.

[3]  Ho-Chul Shin,et al.  Blockade of K+ and Ca2+ channels by azole antifungal agents in neonatal rat ventricular myocytes. , 2012, Biological & pharmaceutical bulletin.

[4]  U. Hass,et al.  Persistent developmental toxicity in rat offspring after low dose exposure to a mixture of endocrine disrupting pesticides. , 2012, Reproductive toxicology.

[5]  Arjen Lommen,et al.  Screening for modulatory effects on steroidogenesis using the human H295R adrenocortical cell line: a metabolomics approach. , 2012, Chemical research in toxicology.

[6]  T. Zacharewski,et al.  BLTK1 murine Leydig cells: a novel steroidogenic model for evaluating the effects of reproductive and developmental toxicants. , 2012, Toxicological sciences : an official journal of the Society of Toxicology.

[7]  A. H. Maia,et al.  Prenatal epoxiconazole exposure effects on rat postnatal development. , 2012, Birth defects research. Part B, Developmental and reproductive toxicology.

[8]  Julie Boberg,et al.  Identification of Cumulative Assessment Groups of Pesticides , 2012 .

[9]  B. Halling‐Sørensen,et al.  H295R cells as a model for steroidogenic disruption: a broader perspective using simultaneous chemical analysis of 7 key steroid hormones. , 2012, Toxicology in vitro : an international journal published in association with BIBRA.

[10]  N. Z. Zur Nieden,et al.  Embryonic stem cell test: stem cell use in predicting developmental cardiotoxicity and osteotoxicity. , 2012, Methods in molecular biology.

[11]  E. Menegola,et al.  Is the amphibian X. laevis WEC a good alternative method to rodent WEC teratogenicity assay? The example of the three triazole derivative fungicides Triadimefon, Tebuconazole, Cyproconazole. , 2011, Reproductive toxicology.

[12]  A. Piersma,et al.  Comparison of the mouse Embryonic Stem cell Test, the rat Whole Embryo Culture and the Zebrafish Embryotoxicity Test as alternative methods for developmental toxicity testing of six 1,2,4-triazoles. , 2011, Toxicology and applied pharmacology.

[13]  A. Luch,et al.  Defined culture medium for stem cell differentiation: applicability of serum-free conditions in the mouse embryonic stem cell test. , 2011, Toxicology in vitro : an international journal published in association with BIBRA.

[14]  Andrea E M Seiler,et al.  The validated embryonic stem cell test to predict embryotoxicity in vitro , 2011, Nature Protocols.

[15]  Aldert H Piersma,et al.  Relative embryotoxicity of two classes of chemicals in a modified zebrafish embryotoxicity test and comparison with their in vivo potencies. , 2011, Toxicology in vitro : an international journal published in association with BIBRA.

[16]  J. Pennings,et al.  Concentration-dependent gene expression responses to flusilazole in embryonic stem cell differentiation cultures. , 2011, Toxicology and applied pharmacology.

[17]  峯島 浩.  Malformation spectrum induced by ketoconazole after single administration to pregnant rats during the critical period : comparison with vitamin A-induced malformation spectrum , 2011 .

[18]  A. Vinggaard,et al.  Endocrine disrupting effects in vitro of conazole antifungals used as pesticides and pharmaceuticals. , 2010, Reproductive toxicology.

[19]  E. Menegola,et al.  Are azole fungicides a teratogenic risk for human conceptus? , 2010, Toxicology letters.

[20]  N. Cedergreen,et al.  Mixture effects of imidazole fungicides on cortisol and aldosterone secretion in human adrenocortical H295R cells. , 2010, Toxicology.

[21]  P. West,et al.  Predicting human developmental toxicity of pharmaceuticals using human embryonic stem cells and metabolomics. , 2010, Toxicology and applied pharmacology.

[22]  R. Regal,et al.  Characterization of the androgen‐sensitive MDA‐kb2 cell line for assessing complex environmental mixtures , 2010, Environmental toxicology and chemistry.

[23]  C. Nellemann,et al.  Mixture effects of endocrine disrupting compounds in vitro. , 2010, International journal of andrology.

[24]  F. Marotta,et al.  Molecular aspects of azoles-induced teratogenesis , 2010, Expert opinion on drug metabolism & toxicology.

[25]  Xiaowei Zhang,et al.  Assessment of chemical effects on aromatase activity using the H295R cell line , 2010, Environmental science and pollution research international.

[26]  S. Pecific.  Proposal for Harmonised Classification and Labelling Based on Regulation ( EC ) No 1272 / 2008 ( CLP Regulation ) , Annex VI , Part 2 Substance Name : 8 : 2 , 2012 .

[27]  Aldert Piersma,et al.  A Review of the Implementation of the Embryonic Stem Cell Test (EST) , 2009, Alternatives to laboratory animals : ATLA.

[28]  V. C. S. Amaral,et al.  Prednisone reduces ketoconazole-induced skeletal defects in rat fetuses , 2009, Archives of Toxicology.

[29]  D. Dix,et al.  Inhibition of Rat and Human Steroidogenesis by Triazole Antifungals , 2009, Systems biology in reproductive medicine.

[30]  R. Frydman,et al.  Phthalates Impair Germ Cell Development in the Human Fetal Testis in Vitro without Change in Testosterone Production , 2008, Environmental health perspectives.

[31]  M. Iida,et al.  In vitro screening for aryl hydrocarbon receptor agonistic activity in 200 pesticides using a highly sensitive reporter cell line, DR-EcoScreen cells, and in vivo mouse liver cytochrome P450-1A induction by propanil, diuron and linuron. , 2008, Chemosphere.

[32]  V. C. S. Amaral,et al.  Ketoconazole- and fluconazole-induced embryotoxicity and skeletal anomalies in wistar rats: a comparative study , 2008 .

[33]  S. Bremer,et al.  Embryotoxicity hazard assessment of cadmium and arsenic compounds using embryonic stem cells. , 2008, Toxicology.

[34]  A M Vinggaard,et al.  Endocrine-disrupting properties in vivo of widely used azole fungicides. , 2008, International journal of andrology.

[35]  Eva Bay Wedebye,et al.  Screening of 397 chemicals and development of a quantitative structure--activity relationship model for androgen receptor antagonism. , 2008, Chemical research in toxicology.

[36]  T. Hartung,et al.  First steps in establishing a developmental toxicity test method based on human embryonic stem cells. , 2008, Toxicology in vitro : an international journal published in association with BIBRA.

[37]  Julie Boberg,et al.  Endocrine-disrupting activities in vivo of the fungicides tebuconazole and epoxiconazole. , 2007, Toxicological sciences : an official journal of the Society of Toxicology.

[38]  Daniel L Villeneuve,et al.  Comparison of fathead minnow ovary explant and H295R cell-based steroidogenesis assays for identifying endocrine-active chemicals. , 2007, Ecotoxicology and environmental safety.

[39]  C. Danielsson,et al.  Embryonic cardiac arrhythmia and generation of reactive oxygen species: common teratogenic mechanism for IKr blocking drugs. , 2007, Reproductive toxicology.

[40]  J. Furr,et al.  Sensitivity of fetal rat testicular steroidogenesis to maternal prochloraz exposure and the underlying mechanism of inhibition. , 2007, Toxicological sciences : an official journal of the Society of Toxicology.

[41]  U. Hass,et al.  Effects of azole fungicides on the function of sex and thyroid hormones , 2007 .

[42]  David J Dix,et al.  Disruption of testosterone homeostasis as a mode of action for the reproductive toxicity of triazole fungicides in the male rat. , 2007, Toxicological sciences : an official journal of the Society of Toxicology.

[43]  John L Newsted,et al.  Human adrenocarcinoma (H295R) cells for rapid in vitro determination of effects on steroidogenesis: hormone production. , 2006, Toxicology and applied pharmacology.

[44]  C. Casellas,et al.  Modulation of aromatase activity and mRNA by various selected pesticides in the human choriocarcinoma JEG-3 cell line. , 2006, Toxicology.

[45]  David J Dix,et al.  Effect of conazole fungicides on reproductive development in the female rat. , 2006, Reproductive toxicology.

[46]  Ulla Hass,et al.  Mechanisms of action underlying the antiandrogenic effects of the fungicide prochloraz. , 2006, Toxicology and applied pharmacology.

[47]  Wolfgang Völkel,et al.  Inhibition of human CYP19 by azoles used as antifungal agents and aromatase inhibitors, using a new LC-MS/MS method for the analysis of estradiol product formation. , 2006, Toxicology.

[48]  Andrea E M Seiler,et al.  Use of murine embryonic stem cells in embryotoxicity assays: the embryonic stem cell test. , 2006, Methods in molecular biology.

[49]  E. Menegola,et al.  Dysmorphogenic effects of some fungicides derived from the imidazole on rat embryos cultured in vitro. , 2006, Reproductive toxicology.

[50]  Ulla Hass,et al.  Perinatal exposure to the fungicide prochloraz feminizes the male rat offspring. , 2005, Toxicological sciences : an official journal of the Society of Toxicology.

[51]  L. Gray,et al.  Late Gestational Exposure to the Fungicide Prochloraz Delays the Onset of Parturition and Causes Reproductive Malformations in Male but Not Female Rat Offspring1 , 2005, Biology of reproduction.

[52]  M. Iida,et al.  A novel nonradioactive method for measuring aromatase activity using a human ovarian granulosa-like tumor cell line and an estrone ELISA. , 2004, Toxicological sciences : an official journal of the Society of Toxicology.

[53]  M. Dalgaard,et al.  The combined antiandrogenic effects of five commonly used pesticides. , 2004, Toxicology and applied pharmacology.

[54]  Susanne Bremer,et al.  Validation of the Embryonic Stem Cell Test in the International ECVAM Validation Study on Three In Vitro Embryotoxicity Tests , 2004, Alternatives to laboratory animals : ATLA.

[55]  G. Held,et al.  Phthalate ester-induced gubernacular lesions are associated with reduced insl3 gene expression in the fetal rat testis. , 2004, Toxicology letters.

[56]  Martin van den Berg,et al.  A comparison of human H295R and rat R2C cell lines as in vitro screening tools for effects on aromatase. , 2004, Toxicology letters.

[57]  Wolfgang Völkel,et al.  Comparative Assessment of the Inhibition of Recombinant Human CYP19 (Aromatase) by Azoles Used in Agriculture and as Drugs for Humans , 2004, Endocrine research.

[58]  A. Vinggaard,et al.  Effects of currently used pesticides in the AhR-CALUX assay: comparison between the human TV101L and the rat H4IIE cell line. , 2003, Toxicology.

[59]  M. Dalgaard,et al.  Antiandrogenic effects in vitro and in vivo of the fungicide prochloraz. , 2002, Toxicological sciences : an official journal of the Society of Toxicology.

[60]  Martin van den Berg,et al.  Induction and inhibition of aromatase (CYP19) activity by various classes of pesticides in H295R human adrenocortical carcinoma cells. , 2002, Toxicology and applied pharmacology.

[61]  Aldert Piersma,et al.  The ECVAM International Validation Study on In Vitro Embryotoxicity Tests: Results of the Definitive Phase and Evaluation of Prediction Models , 2002, Alternatives to laboratory animals : ATLA.

[62]  Anne Marie Vinggaard,et al.  Effects of currently used pesticides in assays for estrogenicity, androgenicity, and aromatase activity in vitro. , 2002, Toxicology and applied pharmacology.

[63]  S. D. Turner,et al.  ASSESSING PREGNANCY RISKS OF AZOLE ANTIFUNGALS USING A HIGH THROUGHPUT AROMATASE INHIBITION ASSAY , 2002, Endocrine research.

[64]  R. Smialowicz,et al.  The effects of perinatal tebuconazole exposure on adult neurological, immunological, and reproductive function in rats. , 2001, Toxicological sciences : an official journal of the Society of Toxicology.

[65]  C. Rider,et al.  The fungicide propiconazole interferes with embryonic development of the crustacean Daphnia magna , 2001, Environmental toxicology and chemistry.

[66]  S. D. Turner,et al.  A high-throughput screen to identify inhibitors of aromatase (CYP19). , 2000, Analytical biochemistry.

[67]  A M Vinggaard,et al.  Screening of selected pesticides for inhibition of CYP19 aromatase activity in vitro. , 2000, Toxicology in vitro : an international journal published in association with BIBRA.

[68]  J. Laskey,et al.  Ketoconazole impairs early pregnancy and the decidual cell response via alterations in ovarian function. , 1997, Fundamental and applied toxicology : official journal of the Society of Toxicology.

[69]  B. Schilter,et al.  Validation of whole chick embryo cultures, whole rat embryo cultures and aggregating embryonic brain cell cultures using six pairs of coded compounds. , 1993, Toxicology in vitro : an international journal published in association with BIBRA.

[70]  R. De Coster,et al.  Comparative effects of ketoconazole on rat, dog and human testicular steroidogenesis. , 1989, Journal of enzyme inhibition.

[71]  M. Levell,et al.  Structure-activity relationships of the inhibition of human placental aromatase by imidazole drugs including ketoconazole. , 1988, Journal of steroid biochemistry.

[72]  C. Rosenfeld,et al.  Ovine placental aromatase: studies of activity levels, kinetic characteristics and effects of aromatase inhibitors. , 1987, Journal of steroid biochemistry.

[73]  J. Ian Mason,et al.  Imidazole antimycotics: Selective inhibitors of steroid aromatization and progesterone hydroxylation , 1987, Steroids.

[74]  M. Levell,et al.  Inhibition of rat testicular 17 alpha-hydroxylase and 17,20-lyase activities by anti-androgens (flutamide, hydroxyflutamide, RU23908, cyproterone acetate) in vitro. , 1987, Journal of steroid biochemistry.

[75]  R. Bechter,et al.  Teratogenicity in vitro-A comparative study of four antimycotic drugs using the whole-embryo culture system. , 1987, Toxicology in vitro : an international journal published in association with BIBRA.

[76]  J. Mason,et al.  Imidazole antimycotics: inhibitors of steroid aromatase. , 1985, Biochemical pharmacology.

[77]  E. Nieschlag,et al.  Effect of ketoconazole and other imidazole fungicides on testosterone biosynthesis. , 1984, Acta endocrinologica.

[78]  D. Stevens,et al.  Ketoconazole blocks testosterone synthesis. , 1982, Archives of internal medicine.

[79]  N. Schwartz,et al.  Uncoupling of vaginal opening and the first ovulation--an indication of an alteration in the pituitary-gonadal axis. , 1977, Biology of Reproduction.