90357-06-5LKJPYSCBVHEWIU-UHFFFAOYSA-NLKJPYSCBVHEWIU-UHFFFAOYSA-N
BicalutamideCasodex
CDX
Propanamide, N-[4-cyano-3-(trifluoromethyl)phenyl]-3-[(4-fluorophenyl)sulfonyl]-2-hydroxy-2-methyl-
DTXSID2022678427-51-0UWFYSQMTEOIJJG-FDTZYFLXSA-NUWFYSQMTEOIJJG-FDTZYFLXSA-N
Cyproterone acetate3'H-Cyclopropa[1,2]pregna-1,4,6-triene-3,20-dione, 17-(acetyloxy)-6-chloro-1,2-dihydro-, (1β,2β)-
1,2α-Methylene-6-chloro-17α-acetoxy-4,6-pregnadiene-3,20-dione
1,2α-Methylene-6-chloro-pregna-4,6-diene-3,20-dione 17α-acetate
1,2α-Methylene-6-chloro-Δ4,6-pregnadien-17α-ol-3,20-dione acetate
17-acetate de 6-chloro-1-β,2-β-dihydro-17-hydroxy-3'H-cyclopropa[1,2]pregna-1,4,6-triene-3,20-dione
17-acetato de 6-cloro-1-β,2-β-dihidro-17-hidroxi-3'H-ciclopropa[1,2]pregna-1,4,6-trieno-3,20-diona
17α-Acetoxy-6-chloro-1α,2α-methylenepregna-4,6-diene-3,20-dione
3'H-Cyclopropa[1,2]pregna-1,4,6-triene-3,20-dione
3'H-Cyclopropa[1,2]pregna-1,4,6-triene-3,20-dione, 6-chloro-1β,2β-dihydro-17-hydroxy-, acetate
6-Chlor-1-β,2-β-dihydro-17-hydroxy-3'H-cyclopropa[1,2]pregna-1,4,6-trien-3,20-dion-17-acetat
6-Chloro-1,2α-methylene-17α-hydroxy-Δ6-progesterone acetate
6-Chloro-1,2α-methylene-6-dehydro-17α-hydroxyprogesterone acetate
6-Chloro-17-hydroxy-1α,2α-methylenepregna-4,6-diene-3,20-dione acetate
6-chloro-1-β,2-β-dihydro-17-hydroxy-3'H-cyclopropa[1,2]pregna-1,4,6-triene-3,20-dione 17-acetate
Androcur
Cyprostat
Cyproterone 17-O-acetate
Cyproterone 17α-acetate
Cyproviron
NSC 81430
Pregna-4,6-diene-3,20-dione, 6-chloro-17-hydroxy-1α,2α-methylene-, acetate
DTXSID5020366133855-98-8ZMYFCFLJBGAQRS-UHFFFAOYNA-NZMYFCFLJBGAQRS-UHFFFAOYSA-N
EpoxiconazoleDTXSID104037213311-84-7MKXKFYHWDHIYRV-UHFFFAOYSA-NMKXKFYHWDHIYRV-UHFFFAOYSA-N
FlutamidePropanamide, 2-methyl-N-[4-nitro-3-(trifluoromethyl)phenyl]-
4-Nitro-3-(trifluoromethyl)isobutyranilide
4'-Nitro-3'-trifluoromethylisobutyranilide
Eulexin
Flucinom
Flutamid
flutamida
m-Propionotoluidide, α,α,α-trifluoro-2-methyl-4'-nitro-
N-(Isopropylcarbonyl)-4-nitro-3-trifluoromethylaniline
Niftholide
Niftolide
NSC 147834
NSC 215876
DTXSID703200485509-19-9FQKUGOMFVDPBIZ-UHFFFAOYSA-NFQKUGOMFVDPBIZ-UHFFFAOYSA-N
FlusilazoleNuStar
DTXSID302423567747-09-5TVLSRXXIMLFWEO-UHFFFAOYSA-NTVLSRXXIMLFWEO-UHFFFAOYSA-N
Prochloraz1H-Imidazole-1-carboxamide, N-propyl-N-[2-(2,4,6-trichlorophenoxy)ethyl]-
BTS 40542-7877
N-propil-N-[2-(2,4,6-triclorofenoxi)etil]-1H-imidazol-1-carboxamida
N-propyl-N-[2-(2,4,6-trichlorophenoxy)ethyl]-1H-imidazole-1-carboxamide
N-Propyl-N-[2-(2,4,6-trichlorophenoxy)ethyl-1H-imidazole-1-carboxamide
N-Propyl-N-[2-(2,4,6-trichlorphenoxy)ethyl]-1H-imidazol-1-carboxamid
Plocloraz
Prelude
Sportak
Sportake
DTXSID402427060207-90-1STJLVHWMYQXCPB-UHFFFAOYNA-NSTJLVHWMYQXCPB-UHFFFAOYSA-N
Propiconazoleppz
1H-1,2,4-Triazole, 1-[[2-(2,4-dichlorophenyl)-4-propyl-1,3-dioxolan-2-yl]methyl]-
(.+-.)-1-[2-(2,4-dichlorophenyl)-4-propyl-1,3-dioxolan-2-yl-methyl]-1H-1,2,4-triazole
(.+-.)-1-[2-(2,4-dichlorophenyl)-4-propyl-1,3-dioxolan-2-ylmethyl]-1H-1,2,4-triazole
1-[[2-(2,4-dichlorophenyl)-4-propyl-1,3-dioxolan-2-yl]methyl]-1H-1,2,4-triazole
1-[[2-(2,4-dichlorophenyl)-4-propyl-1,3-dioxolane-2-yl]methyl]-1H-1,2,4-triazole
1-[[2-(2,4-Dichlorphenyl)-4-propyl-1,3-dioxolan-2-yl]methyl]-1H-1,2,4-triazol
1-[[2-(2,4-diclorofenil)-4-propil-1,3-dioxolan-2-il]metil]-1H-1,2,4-triazol
Bamper 25EC
Banner Maxx
Cane Sett Treatment
Fertilome Liquid Systemic Fungicide
Microban PZ
Microban S 2140
Mycostat P
Proconazole
PROPICONAZOL
Tilt Premium
Wocosen Technical
Wocosin
Wocosin 50TK
DTXSID8024280131983-72-7PPDBOQMNKNNODG-UHFFFAOYNA-NPPDBOQMNKNNODG-UHFFFAOYSA-N
Triticonazole5-[(4-Chlorophenyl)methylene]-2,2-dimethyl-1-(1H-1,2,4-triazol-1-ylmethyl)cyclopentanol
DTXSID0032655107534-96-3PXMNMQRDXWABCY-UHFFFAOYNA-NPXMNMQRDXWABCY-UHFFFAOYSA-N
Tebuconazole1H-1,2,4-Triazole-1-ethanol, .alpha.-(2-(4-chlorophenyl)ethyl)-.alpha.
+-
1H-1,2,4-Triazole-1-ethanol, α-[2-(4-chlorophenyl)ethyl]-α-(1,1-dimethylethyl)-
(.+-.)-Tebuconazole
1-(4-Chlorophenyl)-4,4-dimethyl-3-(1,2,4-triazol-1-ylmethyl)pentan-3-ol
1H-1,2,4-Triazole-1-ethanol, α-[2-(4-chlorophenyl)ethyl]-α-(1,1-dimethylethyl)-, (.+-.)-
1H-1,2,4-Triazole-1-ethanol,α-[2-(4-chlorophenyl) ethyl]-α-(1,1-dimethylethyl)-, (.+-.)-
BAY-HWG 1608
ETHANOL, α-[2-(4-CHLOROPHENYL)ETHYL]-α- (1,1-DIMETHYLETHYL)-1H-1,2,4-TRIAZOLE
Ethyltrianol
Etiltrianol
Fenetrazole
Folicur
Microban S 2142
Microban TZ
Preventol A 8
TEBUCONAZOL
Tebuconazole Resp. HWG 1608
Terbutrazole
α-[2-(4-Chlorophenyl)-ethyl]-α-(1,1-dimethylethyl)-1H-1,2,4-triazole-1-ethanol
α-[2-(4-chlorophenyl)ethyl]-α-(1,1-dimethylethyl)-1H-1,2,4-triazole-1-ethanol
α-tert-Butyl-α-(p-chlorophenethyl)-1H-1,2,4-triazole-1-ethanol
DTXSID903211350471-44-8FSCWZHGZWWDELK-UHFFFAOYNA-NFSCWZHGZWWDELK-UHFFFAOYSA-N
Vinclozolin2,4-Oxazolidinedione, 3-(3,5-dichlorophenyl)-5-ethenyl-5-methyl-
(.+-.)-Vinclozolin
BAS 352-04F
N-3,5-Dichlorophenyl-5-methyl-5-vinyl-1,3-oxazolidine-2,4-dione
N-3,5-Dichlorophenyl-5-methyl-5-vinyloxazolidine-2,4-dione
N-3,5-Dichlorphenyl-5-methyl-5-vinyl-1,3-oxazolidin-2,4-dion
N-3,5-diclorofenil-5-metil-5-vinil-1,3-oxazolidina-2,4-diona
Ornalin
Ranilan
Ronilan
Ronilan 50WP
DTXSID4022361PR:000004191androgen receptorGO:0010468regulation of gene expression2decreasedBicalutamide2020-08-07T06:55:532020-08-07T06:55:53Cyproterone acetate2020-05-17T10:13:282020-05-17T10:13:28Epoxiconazole2020-05-16T11:35:442020-05-16T11:35:44Flutamide2016-11-29T18:42:272016-11-29T18:42:27Flusilazole2020-05-16T11:15:342020-05-16T11:15:34Prochloraz2016-11-29T18:42:222016-11-29T18:42:22Propiconazole2017-05-17T13:18:072017-05-17T13:18:07Stressor:286 Tebuconazole2020-08-07T07:00:532020-08-07T07:00:53Triticonazole2020-05-16T11:02:072020-05-16T11:09:42Vinclozalin2016-11-29T18:42:272016-11-29T18:42:27Mercaptobenzole2016-11-29T18:42:262016-11-29T18:42:26Tebuconazole2017-05-17T13:17:142017-05-17T13:17:14Vinclozolin2020-05-14T11:28:312020-05-14T11:28:31WikiUser_28Vertebrates10116rats10090mouse10116ratDecrease, androgen receptor activationDecrease, AR activationTissue<p><span style="font-size:11pt">This KE refers to decreased activation of the androgen receptor (AR) as occurring in complex biological systems such as tissues and organs in vivo. It is thus considered distinct from KEs describing either blocking of AR or decreased androgen synthesis.</span></p>
<p style="text-align:justify"><span style="font-size:11pt">The AR is a nuclear transcription factor with canonical AR activation regulated by the binding of the androgens such as testosterone or dihydrotestosterone (DHT). Thus, AR activity can be decreased by reduced levels of steroidal ligands (testosterone, DHT) or the presence of compounds interfering with ligand binding to the receptor <span style="color:black">(Davey & Grossmann, 2016; Gao et al., 2005)</span>.</span></p>
<p style="text-align:justify"><span style="font-size:11pt">In the inactive state, AR is sequestered in the cytoplasm of cells by molecular chaperones. In the classical (genomic) AR signaling pathway, AR activation causes dissociation of the chaperones, AR dimerization and translocation to the nucleus to modulate gene expression. AR binds to the androgen response element <span style="color:black">(Davey & Grossmann, 2016; Gao et al., 2005)</span>. AR does not, however, act alone in regulating gene transcription, but together with other co-factors that may differ between cells and tissues and life stages. In this way, the functional consequence of AR activation is cell- and tissue-dependent. </span></p>
<p style="text-align:justify"><span style="font-size:11pt">Ligand-bound AR may also associate with cytoplasmic and membrane-bound proteins to initiate cytoplasmic signaling pathways with other functions than the nuclear pathway. Non-genomic AR signaling includes association with Src kinase to activate MAPK/ERK signaling and activation of the PI3K/Akt pathway. Decreased AR activity may therefore be a decrease in the genomic and/or non-genomic AR signaling pathways <span style="color:black">(Leung & Sadar, 2017)</span>.</span></p>
<p><span style="font-size:11pt">This KE specifically focuses on decreased <em>in vivo</em> activation, with most methods that can be used to measure AR activity carried out <em>in vitro</em>. They provide indirect information about the KE and are described in lower tier MIE/KEs (see MIE/KE-26 for AR antagonism, KE-1690 for decreased T levels and KE-1613 for decreased dihydrotestosterone levels). In this way, this KE is a placeholder for tissue-specific responses to AR activation or inactivation that will depend on the adverse outcome (AO) for which it is included. </span></p>
<p style="text-align:justify"><span style="font-size:11pt">It should be mentioned that the Rapid Androgen Disruption Activity Reporter (RADAR) assay included in OECD test guideline no. 251 detects AR antagonism in vivo in fish (OECD 2022).</span></p>
<p><span style="font-size:11pt">This KE is considered broadly applicable across vertebrate taxa as all vertebrate animals express the AR in numerous cells and tissues where it regulates gene transcription required for developmental processes and functions. </span></p>
HighMixedHighDuring development and at adulthoodHigh<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Davey, R. A., & Grossmann, M. (2016). Androgen Receptor Structure, Function and Biology: From Bench to Bedside. <em>The Clinical Biochemist. Reviews</em>, <em>37</em>(1), 3–15.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Gao, W., Bohl, C. E., & Dalton, J. T. (2005). Chemistry and structural biology of androgen receptor. <em>Chemical Reviews</em>, <em>105</em>(9), 3352–3370. https://doi.org/10.1021/cr020456u</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Hutson, J. M. (1985). A biphasic model for the hormonal control of testicular descent. <em>The Lancet</em>, <em>24</em>, 419–421. https://doi.org/https://doi.org/10.1016/S0140-6736(85)92739-4</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Kaftanovskaya, E. M., Huang, Z., Barbara, A. M., de Gendt, K., Verhoeven, G., Gorlov, I. P., & Agoulnik, A. I. (2012). Cryptorchidism in mice with an androgen receptor ablation in gubernaculum testis. <em>Molecular Endocrinology</em>, <em>26</em>(4), 598–607. https://doi.org/10.1210/me.2011-1283</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Lee, S. H., Hong, K. Y., Seo, H., Lee, H. S., & Park, Y. (2021). Mechanistic insight into human androgen receptor-mediated endocrine-disrupting potentials by a stable bioluminescence resonance energy transfer-based dimerization assay. <em>Chemico-Biological Interactions</em>, <em>349</em>. https://doi.org/10.1016/j.cbi.2021.109655</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Leung, J. K., & Sadar, M. D. (2017). Non-Genomic Actions of the Androgen Receptor in Prostate Cancer. <em>Frontiers in Endocrinology</em>, <em>8</em>. <a href="https://doi.org/10.3389/fendo.2017.00002" style="color:#0563c1; text-decoration:underline">https://doi.org/10.3389/fendo.2017.00002</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">OECD (2022). Test No. 251: Rapid Androgen Disruption Activity Reporter (RADAR) assay. Paris: OECD Publishing doi:10.1787/da264d82-en.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Pang, T. P. S., Clarke, M. v., Ghasem-Zadeh, A., Lee, N. K. L., Davey, R. A., & MacLean, H. E. (2012). A physiological role for androgen actions in the absence of androgen receptor DNA binding activity. <em>Molecular and Cellular Endocrinology</em>, <em>348</em>(1), 189–197. https://doi.org/10.1016/j.mce.2011.08.017</span></span></p>
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<p> </p>
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2019-04-10T05:04:182023-10-19T07:21:18Altered, Transcription of genes by the androgen receptorAltered, Transcription of genes by the ARTissue<p><span style="font-size:11pt"><span style="font-size:12.0pt">This KE refers to transcription of genes by the androgen receptor (AR) as occurring in complex biological systems such as tissues and organs <em>in vivo</em>.</span></span></p>
<p><u>The Androgen Receptor and its function</u></p>
<p><span style="font-size:12.0pt">The AR belongs to the steroid hormone nuclear receptor family. It is a ligand-activated transcription factor with three domains: the N-terminal domain, the DNA-binding domain, and the ligand-binding domain with the latter being the most evolutionary conserved (Davey and Grossmann 2016). </span>Androgens <span style="font-size:12.0pt">(such as dihydrotestosterone and testosterone) are AR ligands and </span>act by binding to the AR in androgen-responsive tissues (Davey and Grossmann 2016). Human AR mutations and mouse knockout models have established a fundamental role for AR in masculinization and spermatogenesis (Maclean et al.; Walters et al. 2010; Rana et al. 2014). The AR is also expressed in many other tissues such as bone, muscles, ovaries and within the immune system (Rana et al. 2014).</p>
<p> </p>
<p><u>Altered transcription of genes by the AR as a Key Event</u></p>
<p>Upon activation by ligand-binding, the AR translocates from the cytoplasm to the cell nucleus, dimerizes, binds to androgen response elements in the DNA to modulate gene transcription (Davey and Grossmann 2016). The transcriptional targets vary between cells and tissues, as well as with developmental stages and is also dependent on available co-regulators (Bevan and Parker 1999; Heemers and Tindall 2007). <span style="font-size:12.0pt">It should also be mentioned that the AR can work in other ‘non-canonial’ ways such as non-genomic signaling, and ligand-independent activation (Davey & Grossmann, 2016; Estrada et al, 2003; Jin et al, 2013). </span></p>
<p>A large number of known, and proposed, target genes of AR canonical signaling have been identified by analysis of gene expression following treatments with AR agonists (Bolton et al. 2007; Ngan et al. 2009<span style="font-size:12.0pt">, Jin et al. 2013</span>).</p>
<p>Altered transcription of genes by the AR can be measured by measuring the transcription level of known downstream target genes by RT-qPCR or other transcription analyses approaches, e.g. transcriptomics.</p>
<p>Both the DNA-binding and ligand-binding domains of the AR are highly evolutionary conserved, whereas the transactivation domain show more divergence, which may affect AR-mediated gene regulation across species (Davey and Grossmann 2016). Despite certain inter-species differences, AR function mediated through gene expression is highly conserved, with mutation studies from both humans and rodents showing strong correlation for AR-dependent development and function (Walters et al. 2010). <span style="font-size:12.0pt">Likewise in fish, androgens are important for development of sexual characteristics (Ogino et al., 2014, 2023). One difference that must be mentioned is that in teleost fish, 11-ketotestosterone is the main androgen in addition to testosterone and DHT and that most teleosts have two <em>ar</em> ohnologs, <em>ara</em> and <em>arb</em>, with arb functioning in a similar manner to the AR in other vertebrates (Ogino et al., 2023). </span></p>
<p><span style="font-size:11pt"><span style="font-size:12.0pt">This KE is considered broadly applicable across vertebrate taxa, sex and developmental stages, as all vertebrate animals express the AR in numerous cells and tissues where it regulates gene transcription required for developmental processes and function. </span></span></p>
CL:0000255eukaryotic cellHighMixedHighDuring development and at adulthoodHigh<p>Bevan C, Parker M (1999) The role of coactivators in steroid hormone action. Exp. Cell Res. 253:349–356</p>
<p>Bolton EC, So AY, Chaivorapol C, et al (2007) Cell- and gene-specific regulation of primary target genes by the androgen receptor. Genes Dev 21:2005–2017. doi: 10.1101/gad.1564207</p>
<p>Davey RA, Grossmann M (2016) Androgen Receptor Structure, Function and Biology: From Bench to Bedside. Clin Biochem Rev 37:3–15</p>
<p>Estrada M, Espinosa A, Müller M, Jaimovich E (2003) Testosterone Stimulates Intracellular Calcium Release and Mitogen-Activated Protein Kinases Via a G Protein-Coupled Receptor in Skeletal Muscle Cells. Endocrinology 144:3586–3597. doi: 10.1210/en.2002-0164</p>
<p>Heemers H V., Tindall DJ (2007) Androgen receptor (AR) coregulators: A diversity of functions converging on and regulating the AR transcriptional complex. Endocr. Rev. 28:778–808</p>
<p><span style="font-size:11pt"><span style="font-size:12.0pt">Jin, Hong Jian, Jung Kim, and Jindan Yu. 2013. “Androgen Receptor Genomic Regulation.” Translational Andrology and Urology 2(3):158–77. doi: 10.3978/j.issn.2223-4683.2013.09.01</span></span></p>
<p>Maclean HE, Chu S, Warne GL, Zajact JD Related Individuals with Different Androgen Receptor Gene Deletions</p>
<p>MacLeod DJ, Sharpe RM, Welsh M, et al (2010) Androgen action in the masculinization programming window and development of male reproductive organs. In: International Journal of Andrology. Blackwell Publishing Ltd, pp 279–287</p>
<p>Ngan S, Stronach EA, Photiou A, et al (2009) Microarray coupled to quantitative RT&ndash;PCR analysis of androgen-regulated genes in human LNCaP prostate cancer cells. Oncogene 28:2051–2063. doi: 10.1038/onc.2009.68<span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><a name="_Hlk148352925"></a></span></span></p>
<p><span style="font-size:11pt"><a name="_Hlk148352925"><span style="font-size:12.0pt">Ogino, Y., Ansai, S., Watanabe, E., Yasugi, M., Katayama, Y., Sakamoto, H., et al. </span></a><span style="font-size:12.0pt">(2023). Evolutionary differentiation of androgen receptor is responsible for sexual characteristic development in a teleost fish. <em>Nat. Commun. 2023 141</em> 14, 1–16. doi:10.1038/s41467-023-37026-6.</span></span></p>
<p><span style="font-size:11pt"><span style="font-size:12.0pt">Ogino, Y., Hirakawa, I., Inohaya, K., Sumiya, E., Miyagawa, S., Denslow, N., et al. (2014). Bmp7 and Lef1 Are the Downstream Effectors of Androgen Signaling in Androgen-Induced Sex Characteristics Development in Medaka. </span><em><span style="font-size:12.0pt">Endocrinology</span></em><span style="font-size:12.0pt"> 155, 449–462. doi:10.1210/EN.2013-1507.</span></span></p>
<p>Rana K, Davey RA, Zajac JD (2014) Human androgen deficiency: Insights gained from androgen receptor knockout mouse models. Asian J. Androl. 16:169–177</p>
<p>Walters KA, Simanainen U, Handelsman DJ (2010) Molecular insights into androgen actions in male and female reproductive function from androgen receptor knockout models. Hum Reprod Update 16:543–558. doi: 10.1093/humupd/dmq003</p>
2016-11-29T18:41:232023-10-19T07:02:17Antagonism, Androgen receptorAntagonism, Androgen receptorMolecular<p><u>The androgen receptor (AR) and its function</u></p>
<p><span style="font-size:12.0pt">The AR is a ligand-activated transcription factor belonging to the steroid hormone nuclear receptor family (</span><span style="font-size:11.0pt"><a href="https://aopwiki.org/events/26#_ENREF_1" title="Davey, 2016 #250"><span style="font-size:12.0pt"><span style="color:#337ab7">Davey & Grossmann, 2016</span></span></a></span><span style="font-size:12.0pt">). The AR has three domains: the N-terminal domain, the DNA-binding domain and the ligand-binding domain, with the latter being most evolutionary conserved. </span>Testosterone (T) and the more biologically active dihydrotestosterone (DHT) are endogenous ligands for the AR (<a href="#_ENREF_4" title="MacLean, 1993 #251">MacLean et al, 1993</a>; <a href="#_ENREF_5" title="MacLeod, 2010 #27">MacLeod et al, 2010</a>; <a href="#_ENREF_8" title="Schwartz, 2019 #252">Schwartz et al, 2019</a>). <span style="font-size:12.0pt">In teleost fishes, 11-ketotestosterone is the second main ligand (<a href="#" title="Schuppe et al, 2020">Schuppe et al, 2020</a>).</span> Human AR mutations and mouse knock-out models have established a pivotal role for the AR in masculinization and spermatogenesis (<a href="#_ENREF_9" title="Walters, 2010 #254">Walters et al, 2010</a>). Apart from the essential role for AR in male reproductive development and function (<a href="#_ENREF_9" title="Walters, 2010 #254">Walters et al, 2010</a>), the AR is also expressed in many other tissues and organs such as bone, muscles, ovaries, and the immune system (<a href="#_ENREF_7" title="Rana, 2014 #253">Rana et al, 2014</a>). </p>
<p><u>AR antagonism as Key Event</u></p>
<p>The main function of the AR is to activate gene transcription in cells. Canonical signaling occurs by ligands (androgens) binding to AR in the cytoplasm which results in translocation to the cell nucleus, receptor dimerization and binding to specific regulatory DNA sequences (<a href="#_ENREF_2" title="Heemers, 2007 #255">Heemers & Tindall, 2007</a>). The gene targets regulated by AR activation depends on cell/tissue type and what stage of development activation occur, and is, for instance, dependent on available co-factors. Apart from the canonical signaling pathway, AR can also <span style="font-size:12.0pt">initiate cytoplasmic signaling pathways with other functions than the nuclear pathway,</span> for instance rapid change in cell function by ion transport changes (<a href="#_ENREF_3" title="Heinlein, 2002 #256">Heinlein & Chang, 2002</a>) <span style="font-size:12.0pt">and association with Src kinase to activate MAPK/ERK signaling and activation of the PI3K/Akt pathway (<a href="#" title="Leung & Sadar, 2017">Leung & Sadar, 2017</a>)</span>. </p>
<p>AR antagonism can be measured in vitro by transient or stable transactivation assays to evaluate nuclear receptor activation. There is already a validated assay for AR (ant)agonism adopted by the OECD, Test No. 458: <em>Stably Transfected Human Androgen Receptor Transcriptional Activation Assay for Detection of Androgenic Agonist and Antagonist Activity of Chemicals </em>(<a href="#_ENREF_13" title="OECD, 2016 #257">OECD, 2016</a>). The stably transfected AR-EcoScreen<sup>TM</sup> cells (<a href="#_ENREF_15" title="Satoh, 2004 #280">Satoh et al, 2004</a>) should be used for the assay and are freely available from the Japanese Collection of Research Bioresources (JCRB) Cell Bank under reference number JCRB1328.</p>
<p>Other assays include the AR-CALUX reporter gene assay that is derived from human U2-OS cells stably transfected with the human AR and an AR responsive reporter gene (<a href="#" title="Sonneveld et al, 2004">Sonneveld et al, 2004</a>; <a href="#_ENREF_18" title="van der Burg, 2010 #261">van der Burg et al, 2010</a>), various transiently transfected reporter cell lines (<a href="#_ENREF_10" title="Körner, 2004 #282">Körner et al, 2004</a>), and more.</p>
<p><span style="font-size:11.0pt">The recently developed AR dimerization assay provides an assay with an improved ability to measure potential stressor-mediated disruption of dimerization/activation (</span><span style="font-size:11.0pt"><a href="#_ENREF_11" title="Lee, 2021 #288">Lee et al, 2021</a></span><span style="font-size:11.0pt">).</span></p>
<p><span style="font-size:11.0pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:16px">The Rapid Androgen Disruption Activity Reporter (RADAR) assay included in OECD test guideline no. 251 detects AR antagonism in vivo in fish (<a href="#">OECD 2022</a>).</span> </span></span></p>
<p>Both the DNA-binding and ligand-binding domains of the AR are highly evolutionary conserved, whereas the transactivation domain show more divergence which may affect AR-mediated gene regulation across species (<a href="#_ENREF_1" title="Davey, 2016 #250">Davey & Grossmann, 2016</a>). Despite certain inter-species differences, AR function mediated through gene expression is highly conserved, with mutations studies from both humans and rodents showing strong correlation for AR-dependent development and function (<a href="#_ENREF_9" title="Walters, 2010 #254">Walters et al, 2010</a>). <span style="font-size:11pt"><span style="font-size:12.0pt">Likewise in fish, androgens are important for development of sexual characteristics (Ogino et al., 2014, 2023). One difference that must be mentioned is that in teleost fish, 11-ketotestosterone is the main androgen in addition to testosterone and DHT and that most teleosts have two <em>ar</em> ohnologs, <em>ara</em> and <em>arb</em>, with arb functioning in a similar manner to the AR in other vertebrates (Ogino et al., 2023).</span></span></p>
<p>This KE is applicable for both sexes, across developmental stages into adulthood, in numerous cells and tissues and across vertebrate taxa</p>
CL:0000255eukaryotic cellHighMixedHighDuring development and at adulthoodHigh<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_2">Davey RA, Grossmann M (2016) Androgen Receptor Structure, Function and Biology: From Bench to Bedside. <em>Clin Biochem Rev</em> <strong>37:</strong> 3-15</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_6">Heemers HV, Tindall DJ (2007) Androgen receptor (AR) coregulators: a diversity of functions converging on and regulating the AR transcriptional complex. <em>Endocr Rev</em> <strong>28:</strong> 778-808</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_7">Heinlein CA, Chang C (2002) The roles of androgen receptors and androgen-binding proteins in nongenomic androgen actions. <em>Mol Endocrinol</em> <strong>16:</strong> 2181-2187</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_10">Körner W, Vinggaard AM, Térouanne B, Ma R, Wieloch C, Schlumpf M, Sultan C, Soto AM (2004) Interlaboratory comparison of four in vitro assays for assessing androgenic and antiandrogenic activity of environmental chemicals. <em>Environ Health Perspect</em> <strong>112:</strong> 695-702</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_11">Lee SH, Hong KY, Seo H, Lee HS, Park Y (2021) Mechanistic insight into human androgen receptor-mediated endocrine-disrupting potentials by a stable bioluminescence resonance energy transfer-based dimerization assay. <em>Chem Biol Interact</em> <strong>349:</strong> 109655</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a id="_ENREF_23" name="_ENREF_23">Leung, J. K., & Sadar, M. D. (2017). Non-Genomic Actions of the Androgen Receptor in Prostate Cancer. <em>Frontiers in Endocrinology</em>, <em>8</em>. https://doi.org/10.3389/fendo.2017.00002</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_12">MacLean HE, Chu S, Warne GL, Zajac JD (1993) Related individuals with different androgen receptor gene deletions. <em>J Clin Invest</em> <strong>91:</strong> 1123-1128</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_13">MacLeod DJ, Sharpe RM, Welsh M, Fisken M, Scott HM, Hutchison GR, Drake AJ, van den Driesche S (2010) Androgen action in the masculinization programming window and development of male reproductive organs. <em>Int J Androl</em> <strong>33:</strong> 279-287</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_14">OECD. (2016) Test No. 458: Stably Transfected Human Androgen Receptor Transcriptional Activation Assay for Detection of Androgenic Agonist and Antagonist Activity of Chemicals. <em>OECD Guidelines for the Testing of Chemicals, Section 4</em>, Paris.</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">OECD (2022). Test No. 251: <a name="_Hlk148359154">Rapid Androgen Disruption Activity Reporter (RADAR) assay</a>. Paris: OECD Publishing doi:10.1787/da264d82-en.</span></span></p>
<p><span style="font-size:14px"><a name="_Hlk148353027">Ogino, Y., Ansai, S., Watanabe, E., Yasugi, M., Katayama, Y., Sakamoto, H., et al. </a>(2023). Evolutionary differentiation of androgen receptor is responsible for sexual characteristic development in a teleost fish. <em>Nat. Commun. 2023 141</em> 14, 1–16. doi:10.1038/s41467-023-37026-6.</span></p>
<p><span style="font-size:14px">Ogino, Y., Hirakawa, I., Inohaya, K., Sumiya, E., Miyagawa, S., Denslow, N., et al. (2014). Bmp7 and Lef1 Are the Downstream Effectors of Androgen Signaling in Androgen-Induced Sex Characteristics Development in Medaka. <em>Endocrinology</em> 155, 449–462. doi:10.1210/EN.2013-1507.</span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_15">Rana K, davey RA, Zajac JD (2014) Human androgen deficiency: insights gained from androgen receptor knockout mouse models. <em>Asian J Androl</em> <strong>16:</strong> 169-177</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_16">Satoh K, Ohyama K, Aoki N, Iida M, Nagai F (2004) Study on anti-androgenic effects of bisphenol a diglycidyl ether (BADGE), bisphenol F diglycidyl ether (BFDGE) and their derivatives using cells stably transfected with human androgen receptor, AR-EcoScreen. <em>Food Chem Toxicol</em> <strong>42:</strong> 983-993</a></span></span></p>
<p><a id="_ENREF_22" name="_ENREF_22"><span style="font-size:14px">Schuppe, E. R., Miles, M. C., and Fuxjager, M. J. (2020). Evolution of the androgen receptor: Perspectives from human health to dancing birds. Mol. Cell. Endocrinol. 499, 110577. doi:10.1016/J.MCE.2019.110577 </span></a></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_17">Schwartz CL, Christiansen S, Vinggaard AM, Axelstad M, Hass U, Svingen T (2019) Anogenital distance as a toxicological or clinical marker for fetal androgen action and risk for reproductive disorders. <em>Arch Toxicol</em> <strong>93:</strong> 253-272</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_18">Sonneveld E, Jansen HJ, Riteco JA, Brouwer A, van der Burg B (2005) Development of androgen- and estrogen-responsive bioassays, members of a panel of human cell line-based highly selective steroid-responsive bioassays. <em>Toxicol Sci</em> <strong>83:</strong> 136-148</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_19">van der Burg B, Winter R, Man HY, Vangenechten C, Berckmans P, Weimer M, Witters H, van der Linden S (2010) Optimization and prevalidation of the in vitro AR CALUX method to test androgenic and antiandrogenic activity of compounds. <em>Reprod Toxicol</em> <strong>30:</strong> 18-24</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a id="_ENREF_21" name="_ENREF_21">Walters KA, Simanainen U, Handelsman DJ (2010) Molecular insights into androgen actions in male and female reproductive function from androgen receptor knockout models. <em>Hum Reprod Update</em> <strong>16:</strong> 543-558</a></span></span></p>
2016-11-29T18:41:222023-10-19T06:24:18Nipple retention (NR), increasednipple retention, increasedIndividual<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">In common laboratory strains of rats and mice, females typically have 6 (rats) or 5 (mice) pairs of nipples along the bilateral milk lines. In contrast, male rats and mice do not have nipples. This is unlike e.g., humans where both sexes have 2 nipples <span style="color:black">(Schwartz et al., 2021)</span>.</span></span></span></span></p>
<p><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">In laboratory rats, high levels of dihydrotestosterone (DHT) induce regression of the nipples in males (Imperato-McGinley & Gautier, 1986; Kratochwil, 1977; Kratochwil & Schwartz, 1976). Females, in the absence of this DHT surge, retain their nipples. This relationship has also been shown in numerous rat studies with perinatal exposure to anti-androgenic chemicals <span style="color:black">(Schwartz et al., 2021)</span></span></span>. <span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">Hence, if juvenile male rats and mice possess nipples, it is considered a sign of perturbed androgen action early in life.</span></span></p>
<p><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">This KE was first published by Pedersen et al (2022). </span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">Nipple retention (NR) is visually assessed, ideally on postnatal day (PND) 12/13 <span style="color:black">(OECD, 2018; Schwartz et al., 2021). However, PND 14 is also an accepted stage of examination (OECD, 2013)</span>. Depending on animal strain, the time when nipples become visible can vary, but the assessment of NR in males should be conducted when nipples are visible in their female littermates <span style="color:black">(OECD, 2013)</span>.</span></span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">Nipples are detected as dark spots (or shadows) called areolae, which resemble precursors to a nipple rather than a fully developed nipple. The dark area may or may not display a nipple bud <span style="color:black">(Hass et al., 2007)</span>. Areolae typically emerge along the milk lines of the male pups corresponding to where female pups display nipples. Fur growth may challenge detection of areolae after PND 14/15. Therefore, the NR assessment should be conducted prior to excessive fur growth. Ideally, all pups in a study are assessed on the same postnatal day to minimize variation due to maturation level <span style="color:black">(OECD, 2013)</span>. </span></span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">NR is occasionally observed in controls. Hence, accurate assessment of NR in controls is needed to detect substance-induced effects on masculine development <span style="color:black">(Schwartz et al., 2021)</span>. It is recommended by the OECD guidance documents 43 and 151 to record NR as a quantitative number rather than a qualitative measure (present/absent or yes/no response). This allows for more nuanced analysis of results, e.g., high control values may be recognized <span style="color:black">(OECD, 2013, 2018)</span>. Studies reporting quantitative measures of NR are therefore considered stronger in terms of weight of evidence.</span></span></span></span></p>
<p><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">Reproducibility of NR results is challenged by the measure being a visual assessment prone to a degree of subjectivity. Thus, NR should be assessed and scored blinded to exposure groups and ideally be performed by the same person(s) within the same study.</span></span></p>
<p><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">The applicability domain of NR is limited to male laboratory strains of rats and mice from birth to juvenile age.</span></span></p>
HighMaleHighBirth to < 1 monthHighHigh<p><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">Hass, U., Scholze, M., Christiansen, S., Dalgaard, M., Vinggaard, A. M., Axelstad, M., Metzdorff, S. B., & Kortenkamp, A. (2007). Combined exposure to anti-androgens exacerbates disruption of sexual differentiation in the rat. <em>Environmental Health Perspectives</em>, <em>115</em>(suppl 1), 122–128.</span></span></p>
<p><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">Imperato-McGinley, J., Binienda, Z., Gedney, J., & Vaughan, E. D. (1986). Nipple Differentiation in Fetal Male Rats Treated with an Inhibitor of the Enzyme 5α-Reductase: Definition of a Selective Role for Dihydrotestosterone. <em>Endocrinology</em>, <em>118</em>(1), 132–137.</span></span></p>
<p><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">Kratochwil, K. (1977). Development and Loss of Androgen Responsiveness in the Embryonic Rudiment of the Mouse Mammary Gland. <em>DEVELOPMENTAL BIOLOGY</em>, <em>61</em>, 358–365.</span></span></p>
<p><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">OECD. (2008). Guidance document 43 on mammalian reproductive toxicity testing and assessment. <em>Environment, Health and Safety Publications</em>, <em>16</em>(43).</span></span></p>
<p><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">OECD. (2013). Guidance document supporting OECD test guideline 443 on the extended one-generation reproductive toxicity test. <em>Environment, Health and Safety Publications</em>, <em>10</em>(151).</span></span></p>
<p><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">OECD. (2016a). Test Guideline 421: Reproduction/Developmental Toxicity Screening Test. <em>OECD Guidelines for the Testing of Chemicals</em>, <em>421</em>. </span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"><a href="http://www.oecd.org/termsandconditions/" style="color:#0563c1; text-decoration:underline">http://www.oecd.org/termsandconditions/</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">OECD. (2016b). Test Guideline 422: Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test. <em>OECD Guidelines for the Testing of Chemicals</em>, <em>422</em>. <a href="http://www.oecd.org/termsandconditions/" style="color:#0563c1; text-decoration:underline">http://www.oecd.org/termsandconditions/</a> </span></span></p>
<p><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">OECD. (2018). Test Guideline 443: Extended one-generation reproductive toxicity study. <em>OECD Guidelines for the Testing of Chemicals</em>, <em>443</em>. </span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"><a href="http://www.oecd.org/termsandconditions/" style="color:#0563c1; text-decoration:underline">http://www.oecd.org/termsandconditions/</a></span></span></p>
<p><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">Pedersen, E. B., Christiansen, S., & Svingen, T. (2022). AOP key event relationship report: Linking androgen receptor antagonism with nipple retention. Current Research in Toxicology, 3, 100085.</span></span></p>
<p><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">Schwartz, C. L., Christiansen, S., Hass, U., Ramhøj, L., Axelstad, M., Löbl, N. M., & Svingen, T. (2021). On the Use and Interpretation of Areola/Nipple Retention as a Biomarker for Anti-androgenic Effects in Rat Toxicity Studies. <em>Frontiers in Toxicology</em>, <em>3</em>, 730752.</span></span></p>
2020-05-11T10:07:082023-01-11T05:53:2567d53e7b-68b6-4718-a997-4708ebb349468582eaef-c27c-4a3d-b719-a1154db2f653<p style="text-align:justify"><span style="font-size:11pt">The androgen receptor (AR) is a ligand-activated steroid hormone nuclear receptor <span style="color:black">(Davey & Grossmann, 2016)</span>. In its inactive state, the AR locates to the cytoplasm <span style="color:black">(Roy et al., 2001)</span>. When activated, the AR translocates to the nucleus, dimerizes, and, together with co-regulators, binds to specific DNA regulatory sequences to regulate gene transcription <span style="color:black">(Davey & Grossmann, 2016)</span> (Lamont and Tindall, 2010). This is considered the canonical AR signaling pathway. The AR can also activate non-genomic signalling <span style="color:black">(Jin et al., 2013)</span>. However, this KER focuses on the canonical pathway.</span></p>
<p style="text-align:justify"><span style="font-size:11pt">The two main AR ligands are the androgens testosterone (T) and the more potent dihydrotestosterone (DHT), whereas another main androgen in teleost fishes is 11-ketotestosterone (Schuppe et al., 2020). Androgens bind to the AR to mediate downstream androgenic responses, such as male development and masculinization <span style="color:black">(Rey, 2021; Walters et al., 2010)</span>. Antagonism of the AR would decrease AR activation and therefore the downstream AR-mediated effects. </span></p>
<p style="text-align:justify"><span style="font-size:11pt">The biological plausibility for this KER is considered high.</span></p>
<p style="text-align:justify"><span style="font-size:11pt">The AR belongs to the steroid hormone nuclear receptor family. The AR has 3 main domains essential for its activity, the N-terminal domain, the ligand binding domain, and the DNA binding domain <span style="color:black">(Roy et al., 2001)</span>. Ligands, such as T and DHT, must bind to the ligand binding domain to activate AR allowing it to fulfill its role as a transcription factor. The binding of the ligand induces a change in AR conformation allowing it to translocate to the nucleus and congregate into a subnuclear compartment <span style="color:black">(Marcelli et al., 2006; Roy et al., 2001)</span> homodimerize and bind to the DNA target sequences and regulate transcription of target genes. Regulation of AR target genes is greatly facilitated by numerous co-factors. Active AR signaling is essential for male reproduction and sexual development and is also crucial in several other tissues and organs such as ovaries, the immune system, bones, and muscles <span style="color:black">(Ogino et al., 2011; Prizant et al., 2014; Rey, 2021; William H. Walker, 2021)</span>. </span></p>
<p style="text-align:justify"><span style="font-size:11pt">AR antagonists can compete with or prevent in different ways AR ligand binding, thereby preventing AR activation. Antagonism of the AR can prevent translocation to the nucleus, compartmentalization, dimerization and DNA binding. Consequently, AR cannot regulate transcription of target genes and androgen signalling is disrupted. This can be observed using different AR activation assays such as AR dimerization, translocation, DNA binding or transcriptional activity assays <span style="color:black">(Brown et al., 2023; <em>OECD</em>, 2020)</span><span style="color:black">.</span> </span></p>
<p style="text-align:justify"><span style="font-size:11pt">The empirical evidence for this KER is considered high</span></p>
<p style="text-align:justify"><span style="font-size:11pt">The effects of AR antagonism have been shown in many studies <em>in vivo</em> and <em>in vitro</em>. </span></p>
<p><span style="font-size:11pt">Several stressors can act as antagonists of the AR and lead to decreased AR activation. Some of these are detailed in an AOP key event relationship report by <span style="color:black">(Pedersen et al., 2022)</span> and shown below, exhibiting evidence of dose-concordance:</span></p>
<p> </p>
<p><span style="font-size:11pt"><strong>Stressors</strong></span></p>
<ul>
<li><span style="font-size:11pt">Cyproterone acetate: Using the AR-CALUX reporter assay in antagonism mode, cyproterone acetate showed an IC50 of 7.1 nM <span style="color:black">(Sonneveld, 2005)</span></span></li>
<li><span style="font-size:11pt">Epoxiconazole: Using transiently AR-transfected CHO cells, epoxiconazole showed a LOEC of 1.6 µM and an IC50 of 10 µM <span style="color:black">(Kjærstad et al., 2010)</span>.</span></li>
<li><span style="font-size:11pt">Flutamide: Using the AR-CALUX reporter assay in antagonism mode, flutamide showed an IC50 of 1.3 µM <span style="color:black">(Sonneveld, 2005).</span></span></li>
<li><span style="font-size:11pt">Flusilazole: Using hAR-EcoScreen Assay, triticonazole showed a LOEC for antagonisms of 0.8 µM and an IC50 of 2.8 (±0.1) µM <span style="color:#0563c1"><u><span style="color:black">(Draskau et al., 2019)</span></u></span>.</span></li>
<li><span style="font-size:11pt">Prochloraz: Using transiently AR-transfected CHO cells, prochloraz showed a LOEC of 6.3 µM and an IC50 of 13 µM <span style="color:black">(Kjærstad et al., 2010)</span>.</span></li>
<li><span style="font-size:11pt">Propiconazole: Using transiently AR-transfected CHO cells, propiconazole showed a LOEC of 12.5 µM and an IC50 of 18 µM <span style="color:black">(Kjærstad et al., 2010)</span>.</span></li>
<li><span style="font-size:11pt">Tebuconazole: Using transiently AR-transfected CHO cells, tebuconazole showed a LOEC of 3.1 µM and an IC50 of 8.1 µM <span style="color:black">(Kjærstad et al., 2010)</span>.</span></li>
<li><span style="font-size:11pt">Triticonazole: Using hAR-EcoScreen Assay, triticonazole showed a LOEC for antagonisms of 0.2 µM and an IC50 of 0.3 (±0.01) µM <span style="color:black">(Draskau et al., 2019)</span>.</span></li>
<li><span style="font-size:11pt">Vinclozolin: Using the AR-CALUX reporter assay in antagonism mode, vinclozolin showed an IC50of 1.0 µM<span style="color:black">(Sonneveld, 2005)</span>.”<span style="color:black">(Pedersen et al., 2022)</span></span></li>
</ul>
<p><span style="font-size:11pt"><strong>Other evidence: </strong></span></p>
<p style="text-align:justify"><span style="font-size:11pt">Known AR antagonists are used for treatment of AR-sensitive cancers such as flutamide for prostate cancer (Mahler et al., 1998). </span></p>
<p> </p>
<p> </p>
HighMixedHighDuring development and at adulthoodHigh<p style="text-align:justify"><span style="font-size:11pt">The AR is expressed throughout vertebrate taxa and its DNA and ligand binding domains are highly conserved <span style="color:black">(Davey & Grossmann, 2016).</span> AR activity is important for sexual development and reproduction in both males and females <span style="color:black">(Prizant et al., 2014; Walters et al., 2010)</span>. AR function is required during development, puberty and adulthood. </span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Brown, E. C., Hallinger, D. R., Simmons, S. O., Puig-Castellví, F., Eilebrecht, E., Arnold, L., & Bioscience, P. A. (2023). High-throughput AR dimerization assay identifies androgen disrupting chemicals and metabolites. <em>Front. Toxicol</em>, <em>5</em>, 1134783. https://doi.org/10.3389/ftox.2023.1134783</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Chen, C. D., Welsbie, D. S., Tran, C., Baek, S. H., Chen, R., Vessella, R., Rosenfeld, M. G., & Sawyers, C. L. (2003). A R T I C L E S Molecular determinants of resistance to antiandrogen therapy. <em>NATURE MEDICINE</em>, <em>10</em>(1). https://doi.org/10.1038/nm972</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Dart, D. A., Ashelford, K., & Jiang, W. G. (2020). <em>AR mRNA stability is increased with AR-antagonist resistance via 3′UTR variants</em>. https://doi.org/10.1530/EC-19-0340</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Davey, R. A., & Grossmann, M. (2016). Androgen Receptor Structure, Function and Biology: From Bench to Bedside. In <em>Androgen Receptor Biology Clin Biochem Rev</em> (Vol. 37, Issue 1).</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Draskau, M. K., Boberg, J., Taxvig, C., Pedersen, M., Frandsen, H. L., Christiansen, S., & Svingen, T. (2019). In vitro and in vivo endocrine disrupting effects of the azole fungicides triticonazole and flusilazole. <em>Environmental Pollution</em>, <em>255</em>, 113309. https://doi.org/10.1016/j.envpol.2019.113309</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Jin, H. J., Kim, J., & Yu, J. (2013). Androgen receptor genomic regulation. In <em>Translational Andrology and Urology</em> (Vol. 2, Issue 3, pp. 158–177). AME Publishing Company. https://doi.org/10.3978/j.issn.2223-4683.2013.09.01</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Kang, Z., Pirskanen, A., Jänne, O. A., & Palvimo, J. J. (2002). Involvement of Proteasome in the Dynamic Assembly of the Androgen Receptor Transcription Complex. <em>Journal of Biological Chemistry</em>, <em>277</em>(50), 48366–48371. https://doi.org/10.1074/jbc.M209074200</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Kjærstad, M. B., Taxvig, C., Nellemann, C., Vinggaard, A. M., & Andersen, H. R. (2010). Endocrine disrupting effects in vitro of conazole antifungals used as pesticides and pharmaceuticals. <em>Reproductive Toxicology</em>, <em>30</em>(4), 573–582. <a href="https://doi.org/10.1016/j.reprotox.2010.07.009" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1016/j.reprotox.2010.07.009</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Lamont, K. R., and Tindall, D. J. (2010). Androgen Regulation of Gene Expression. Adv. Cancer Res. 107, 137–162. doi:10.1016/S0065-230X(10)07005-3.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Mahler, C., Verhelst, J., and Denis, L. (1998). Clinical pharmacokinetics of the antiandrogens and their efficacy in prostate cancer. Clin. Pharmacokinet. 34, 405–417. doi:10.2165/00003088-199834050-00005/METRICS.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Marcelli, M., Stenoien, D. L., Szafran, A. T., Simeoni, S., Agoulnik, I. U., Weigel, N. L., Moran, T., Mikic, I., Price, J. H., & Mancini, M. A. (2006). Quantifying effects of ligands on androgen receptor nuclear translocation, intranuclear dynamics, and solubility. <em>Journal of Cellular Biochemistry</em>, <em>98</em>(4), 770–788. <a href="https://doi.org/10.1002/jcb.20593" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1002/jcb.20593</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">OECD (2020). Test No. 458: Stably Transfected Human Androgen Receptor Transcriptional Activation Assay for Detection of Androgenic Agonist and Antagonist Activity of Chemicals. OECD Guide. Paris: OECD Publishing doi:10.1787/9789264264366-en.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Ogino, Y., Miyagawa, S., Katoh, H., Prins, G. S., Iguchi, T., & Yamada, G. (2011). Essential functions of androgen signaling emerged through the developmental analysis of vertebrate sex characteristics. <em>Evolution & Development</em>, <em>13</em>(3), 315–325. https://doi.org/10.1111/j.1525-142X.2011.00482.x</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Pedersen, E. B., Christiansen, S., & Svingen, T. (2022). AOP key event relationship report: Linking androgen receptor antagonism with nipple retention. <em>Current Research in Toxicology</em>, <em>3</em>, 100085. https://doi.org/10.1016/j.crtox.2022.100085</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Prizant, H., Gleicher, N., & Sen, A. (2014). Androgen actions in the ovary: balance is key. <em>Journal of Endocrinology</em>, <em>222</em>(3), R141–R151. https://doi.org/10.1530/JOE-14-0296</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Rey, R. A. (2021). The Role of Androgen Signaling in Male Sexual Development at Puberty. <em>Endocrinology</em>, <em>162</em>(2). https://doi.org/10.1210/endocr/bqaa215</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Roy, A. K., Tyagi, R. K., Song, C. S., Lavrovsky, Y., Ahn, S. C., Oh, T. S., & Chatterjee, B. (2001). Androgen receptor: Structural domains and functional dynamics after ligand-receptor interaction. <em>Annals of the New York Academy of Sciences</em>, <em>949</em>, 44–57. https://doi.org/10.1111/j.1749-6632.2001.tb04001.x</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Sonneveld, E. (2005). Development of Androgen- and Estrogen-Responsive Bioassays, Members of a Panel of Human Cell Line-Based Highly Selective Steroid-Responsive Bioassays. <em>Toxicological Sciences</em>, <em>83</em>(1), 136–148. https://doi.org/10.1093/toxsci/kfi005</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Szafran, A. T., Szwarc, M., Marcelli, M., & Mancini, M. A. (2008). Androgen Receptor Functional Analyses by High Throughput Imaging: Determination of Ligand, Cell Cycle, and Mutation-Specific Effects. <em>PLoS ONE</em>, <em>3</em>(11), e3605. https://doi.org/10.1371/journal.pone.0003605</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Walters, K. A., Simanainen, U., & Handelsman, D. J. (2010). Molecular insights into androgen actions in male and female reproductive function from androgen receptor knockout models. In <em>Human Reproduction Update</em> (Vol. 16, Issue 5, pp. 543–558). Hum Reprod Update. https://doi.org/10.1093/humupd/dmq003</span></span></p>
<p><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">William H. Walker. (2021). Androgen Actions in the Testis and the Regulation of Spermatogenesis. In <em>Advances in Experimental Medicine and Biology: Vol. volume 1381</em> (pp. 175–203).</span></span></p>
2020-05-11T07:39:002023-10-19T08:49:1767d53e7b-68b6-4718-a997-4708ebb349466efacf59-b918-4fa7-9464-a0b0b8360930<p><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">Several chemicals can antagonize the androgen receptor (AR) <em>in vitro</em>, resulting in decreased AR activation. Decreased AR activation can lead to incomplete reproductive development in males, which can be expressed in several ways. One endpoint affected is areola/nipple retention (NR), which <em>in vivo</em> studies have shown to be linked to suppressed AR activation. NR in rat and mouse toxicity studies is considered an adverse effect (i.e., an AO).</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">The biological plausibility of a link between decreased AR activation and increased NR in male rats is high. The relationship is supported by numerous studies showing that several potent AR antagonists <em>in vitro</em> induce NR <em>in vivo</em>. However, in the literature review conducted for this KER, no studies in mice were found to fulfill the inclusion criteria. The present KER is hence exclusively a description of the situation in rats, although it is believed that the link also exists in mice. </span></span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">The AR is activated through binding of either testosterone or dihydrotestosterone (DHT), the latter having the highest affinity for the AR. Upon binding, the AR translocates to the target cell nucleus where it acts as a transcription factor <span style="color:black">(Albert, 2018)</span>.</span></span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">NT has </span></span></span></span><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">been shown to be more dependent on DHT-signaling, which suggests that chemicals inducing increased NR also have a higher affinity for the AR than DHT in order to outcompete DHT for AR binding, although supra-high doses of chemicals with lower AR affinity could be speculated to also outcompete T or DHT. The general principle of higher affinity, however, has been confirmed by <em>in vitro </em>studies <span style="color:black">(Gray et al., 2019; Hass et al., 2012; McIntyre et al., 2000)</span>.</span></span></p>
<p> </p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">Table 2 lists chemical stressors shown to antagonize the AR <em>in vitro</em> as well as causing NR in male rat offspring <em>in vivo.</em> Additional information from the <em>in vivo </em>studies, including the animal species and strain, as well as the doses tested, the dosing period and the time of measurement of NR are specified in this table. The lowest dose yielding a significant increase of retained nipples in male rat pups is defined as the LOAEL. Conversely, the NOAEL represents the highest tested dose yielding no significant increase in NR. Note that the given NOAEL and LOAEL values are highly dependent on study design. Significant values are marked with an asterisk.</span></span></span></span></p>
<p><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">Table 3 shows a list of stressors shown to have AR antagonistic properties <em>in vitro</em> or in other <em>in vivo</em> studies, but for which the doses tested <em>in vivo</em> did not produce a significant effect on NR. In this list, the lowest tested dose is reported, and the NOAEL presents the highest dose tested which produced no statistically significant effect on NR. Apart from the NOAEL, the information given in Table 3 is identical to Table 2.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">A major challenge with NR as a biomarker is the subjectivity of the measure. In juvenile rat pups, nipples are only present as areolae, i.e., dark shadows with or without a nipple bud. This means that the experience of the personnel assessing the presence and number of areolae/nipples can influence the results. Furthermore, the results are likely prone to larger variation if several assessors are used to record NR within the same study. To minimize these sources of uncertainty, assessors must be trained to recognize areolae and not look for fully developed nipples. Moreover, the number of assessors should be limited to one or two, and they should always be blinded to exposure groups. </span></span></span></span></p>
<p><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">Another factor that may affect NR results is the age of the rat pups at the time of assessment. OECD guidelines have standardized the time for measuring occurrence of NR to be optimal at PD 12 or 13, when they are visible in female littermates <span style="color:black">(OECD, 2013)</span>. However, assessment of permanent NR is not included in any international guidelines. Hence, if NR is measured in older offspring, the time of measurement is not consistent between studies and varies between PD 20 and PD 100. Thus, conclusions on whether NR is permanent or not may differ based on study design. This distinction between a transient and a permanent effect is important from a regulatory perspective, since only a permanent effect will be categorized as a malformation according to OECD guidance document 43 <span style="color:black">(OECD, 2008)</span>.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">The quantitative understanding of the relationship between decreased AR activity and NR is challenged by the fact that the potency of AR antagonism <em>in vitro</em> is not proportional to the magnitude of NR observed <em>in vivo </em><span style="color:black">(Gray et al., 2019)</span>. Hence, predicting <em>in vivo</em> effects based on <em>in vitro</em> data is not possible. However, <em>in vitro</em> studies can give indications of which chemicals might exhibit anti-androgenic effects <em>in vivo </em>and should be subject to further testing <span style="color:black">(Hass et al., 2012)</span>. Development of more representative <em>in vitro</em> models is necessary if <em>in vivo</em> tests are to be phased out entirely.</span></span></span></span></p>
HighMaleHighDevelopmentHighHigh<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Alapi, E. M., & Fischer, J. (2006). Table of Selected Analogue Classes. In <em>Analogue-based Drug Discovery</em> (pp. 441–552). Wiley-VCH Verlag GmbH & Co. KGaA. <a href="https://doi.org/10.1002/3527608001.ch23" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1002/3527608001.ch23</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Albert, O. (2018). Antiandrogens. In <em>Encyclopedia of Reproduction</em> (Vol. 1, pp. 594–601). Elsevier. <a href="https://doi.org/10.1016/B978-0-12-801238-3.64380-5" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1016/B978-0-12-801238-3.64380-5</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Barlow, N. J., McIntyre, B. S., & Foster, P. M. D. (2004). Male Reproductive Tract Lesions at 6, 12, and 18 Months of Age Following in Utero Exposure to Di(n-butyl) Phthalate. <em>Toxicologic Pathology</em>, <em>32</em>(1), 79–90. <a href="https://doi.org/10.1080/01926230490265894" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1080/01926230490265894</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Bowman, C. J., Barlow, N. J., Turner, K. J., Wallace, D. G., & Foster, P. M. D. (2003). Effects of in utero exposure to finasteride on androgen-dependent reproductive development in the male rat. <em>Toxicological Sciences</em>, <em>74</em>(2), 393–406. <a href="https://doi.org/10.1093/toxsci/kfg128" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1093/toxsci/kfg128</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Carruthers, C. M., & Foster, P. M. D. (2005). Critical window of male reproductive tract development in rats following gestational exposure to di-n-butyl phthalate. <em>Birth Defects Research (Part B) Developmental and Reproductive Research</em>, <em>74</em>(3), 277–285. <a href="https://doi.org/10.1002/bdrb.20050" style="color:#0563c1; text-decoration:underline"><span style="color:#0563c1">https://doi.org/10.1002/bdrb.20050</span></a></span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Christiansen, S., Axelstad, M., Scholze, M., Johansson, H.K.L., Hass, U., Mandrup, K., Frandsen, H.L., Frederiksen, H., Isling, L.K., Boberg, J. (2020). Grouping of endocrine disrupting chemicals for mixture risk assessment – Evidence from a rat study. <em>Environ Int</em>, 142, 105870. <a href="https://doi.org/10.1016/j.envint.2020.105870" style="color:#0563c1; text-decoration:underline"><span style="color:#0563c1">https://doi.org/10.1016/j.envint.2020.105870</span></a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Christiansen, S., Boberg, J., Axelstad, M., Dalgaard, M., Vinggaard, A. M., Metzdorff, S. B., & Hass, U. (2010). Low-dose perinatal exposure to di(2-ethylhexyl) phthalate induces anti-androgenic effects in male rats. <em>Reproductive Toxicology</em>, <em>30</em>(2), 313–321. <a href="https://doi.org/10.1016/j.reprotox.2010.04.005" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1016/j.reprotox.2010.04.005</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Christiansen, S., Scholze, M., Axelstad, M., Boberg, J., Kortenkamp, A., & Hass, U. (2008). Combined exposure to anti-androgens causes markedly increased frequencies of hypospadias in the rat. <em>International Journal of Andrology</em>, <em>31</em>(2), 241–248. <a href="https://doi.org/10.1111/j.1365-2605.2008.00866.x" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1111/j.1365-2605.2008.00866.x</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Christiansen, S., Scholze, M., Dalgaard, M., Vinggaard, A., Axelstad, M., Kortenkamp, A., & Hass, U. (2009). Synergistic disruption of external male sex organ development by a mixture of four antiandrogens. <em>Environmental Health Perspectives</em>, <em>117</em>(12), 1839–1846. <a href="https://doi.org/https:/doi.org/10.1289/ehp.0900689" style="color:#0563c1; text-decoration:underline">https://doi.org/https://doi.org/10.1289/ehp.0900689</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Clewell, R. A., Thomas, A., Willson, G., Creasy, D. M., & Andersen, M. E. (2013). A dose response study to assess effects after dietary administration of diisononyl phthalate (DINP) in gestation and lactation on male rat sexual development. <em>Reproductive Toxicology</em>, <em>35</em>(1), 70–80. <a href="https://doi.org/10.1016/j.reprotox.2012.07.008" style="color:#0563c1; text-decoration:underline"><span style="color:#0563c1">https://doi.org/10.1016/j.reprotox.2012.07.008</span></a></span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Conley JM, Lambright CS, Evans N, Cardon M, Furr J, Wilson VS, Gray LE Jr (2018). Mixed "Antiandrogenic" Chemicals at Low Individual Doses Produce Reproductive Tract Malformations in the Male Rat. <em>Toxicol Sci</em>. 164(1), 166-178. <a href="https://doi.org/10.1093/toxsci/kfy069" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1093/toxsci/kfy069</a> </span></span></p>
<p style="margin-left:38px"> </p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Conley JM, Lambright CS, Evans N, Cardon M, Medlock-Kakaley E, Wilson VS, Gray LE Jr. (2021). A mixture of 15 phthalates and pesticides below individual chemical no observed adverse effect levels (NOAELs) produces reproductive tract malformations in the male rat. <em>Environ Int</em>. 156, 106615. <a href="https://doi.org/10.1016/j.envint.2021.106615" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1016/j.envint.2021.106615</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Davey, R. A., & Grossmann, M. (2016). Androgen Receptor Structure, Function and Biology: From Bench to Bedside. <em>The Clinical Biochemist. Reviews</em>, <em>37</em>(1), 3–15.</span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Draskau, M. K., Ballegaard, A. S. R., Ramhøj, L., Bowles, J., Svingen, T., & Spiller, C. M. (2022). AOP Key Event Relationship report: Linking decreased retinoic acid levels with disrupted meiosis in developing oocytes. <em>Current Research in Toxicology</em>, <em>3</em>(100069). <a href="https://doi.org/10.1016/j.crtox.2022.100069" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1016/j.crtox.2022.100069</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Draskau, M. K., Boberg, J., Taxvig, C., Pedersen, M., Frandsen, H. L., Christiansen, S., & Svingen, T. (2019). In vitro and in vivo endocrine disrupting effects of the azole fungicides triticonazole and flusilazole. <em>Environmental Pollution</em>, <em>255</em>, 113309. <a href="https://doi.org/10.1016/j.envpol.2019.113309" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1016/j.envpol.2019.113309</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Foster, P. M. D., & Harris, M. W. (2005). Changes in androgen-mediated reproductive development in male rat offspring following exposure to a single oral dose of flutamide at different gestational ages. <em>Toxicological Sciences</em>, <em>85</em>(2), 1024–1032. <a href="https://doi.org/10.1093/toxsci/kfi159" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1093/toxsci/kfi159</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Fussell, K. C., Schneider, S., Buesen, R., Groeters, S., Strauss, V., Melching-Kollmuss, S., & van Ravenzwaay, B. (2015). Investigations of putative reproductive toxicity of low-dose exposures to flutamide in Wistar rats. <em>Archives of Toxicology</em>, <em>89</em>(12), 2385–2402. <a href="https://doi.org/10.1007/s00204-015-1622-6" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1007/s00204-015-1622-6</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Gray, L. E., Furr, J. R., Conley, J. M., Lambright, C. S., Evans, N., Cardon, M. C., Wilson, V. S., Foster, P. M., & Hartig, P. C. (2019). A Conflicted Tale of Two Novel AR Antagonists In Vitro and In Vivo: Pyrifluquinazon Versus Bisphenol C. <em>Toxicological Sciences</em>, <em>168</em>(2), 632–643. <a href="https://doi.org/10.1093/toxsci/kfz010" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1093/toxsci/kfz010</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Gray, L. E., Ostby, J., Furr, J., Price, M., Veeramachaneni, D. N. R., & Parks, L. (2000). Perinatal Exposure to the Phtalates DEHP, BBP, and DINP, but Not DEP, DMP, or DOTP, Alters Sexual Differentiation of the Male Rat. <em>Toxicological Sciences</em>, <em>58</em>, 350–365. <a href="https://doi.org/10.1093/toxsci/58.2.350" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1093/toxsci/58.2.350</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Hass, U., Boberg, J., Christiansen, S., Jacobsen, P. R., Vinggaard, A. M., Taxvig, C., Poulsen, M. E., Herrmann, S. S., Jensen, B. H., Petersen, A., Clemmensen, L. H., & Axelstad, M. (2012). Adverse effects on sexual development in rat offspring after low dose exposure to a mixture of endocrine disrupting pesticides. <em>Reproductive Toxicology</em>, <em>34</em>(2), 261–274. <a href="https://doi.org/10.1016/j.reprotox.2012.05.090" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1016/j.reprotox.2012.05.090</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Hass, U., Scholze, M., Christiansen, S., Dalgaard, M., Vinggaard, A. M., Axelstad, M., Metzdorff, S. B., & Kortenkamp, A. (2007). Combined exposure to anti-androgens exacerbates disruption of sexual differentiation in the rat. <em>Environmental Health Perspectives</em>, <em>115</em>(suppl 1), 122–128. <a href="https://doi.org/10.1289/ehp.9360" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1289/ehp.9360</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Heemers, H. v., & Tindall, D. J. (2007). Androgen Receptor (AR) Coregulators: A Diversity of Functions Converging on and Regulating the AR Transcriptional Complex. <em>Endocrine Reviews</em>, <em>28</em>(7), 778–808. <a href="https://doi.org/10.1210/er.2007-0019" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1210/er.2007-0019</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Heinlein, C. A., & Chang, C. (2002). The Roles of Androgen Receptors and Androgen-Binding Proteins in Nongenomic Androgen Actions. <em>Molecular Endocrinology</em>, <em>16</em>(10), 2181–2187. <a href="https://doi.org/10.1210/me.2002-0070" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1210/me.2002-0070</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Hellwig, J., van Ravenzwaay, B., Mayer, M., & Gembardt, C. (2000). Pre- and postnatal oral toxicity of vinclozolin in Wistar and Long-Evans rats. <em>Regulatory Toxicology and Pharmacology</em>, <em>32</em>(1), 42–50. <a href="https://doi.org/10.1006/rtph.2000.1400" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1006/rtph.2000.1400</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Hotchkiss, A. K., Parks-Saldutti, L. G., Ostby, J. S., Lambright, C., Furr, J., Vandenbergh, J. G., & Gray, L. E. (2004). A mixture of the “antiandrogens” linuron and butyl benzyl phthalate alters sexual differentiation of the male rat in a cumulative fashion. <em>Biology of Reproduction</em>, <em>71</em>(6), 1852–1861. <a href="https://doi.org/10.1095/biolreprod.104.031674" style="color:#0563c1; text-decoration:underline"><span style="color:#0563c1">https://doi.org/10.1095/biolreprod.104.031674</span></a></span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Howdeshell KL, Hotchkiss AK, Gray LE Jr (2017). Cumulative effects of antiandrogenic chemical mixtures and their relevance to human health risk assessment. <em>Int J Hyg Environ Health</em>. 220(2 Pt A), 179-188. <a href="https://doi.org/10.1016/j.ijheh.2016.11.007" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1016/j.ijheh.2016.11.007</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Huliganga, E., Marchetti, F., O’Brien, J. M., Chauhan, V., & Yauk, C. L. (2022). A Case Study on Integrating a New Key Event Into an Existing Adverse Outcome Pathway on Oxidative DNA Damage: Challenges and Approaches in a Data-Rich Area. <em>Frontiers in Toxicology</em>, <em>4</em>(827328). <a href="https://doi.org/10.3389/ftox.2022.827328" style="color:#0563c1; text-decoration:underline">https://doi.org/10.3389/ftox.2022.827328</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Imperato-McGinley, J., Binienda, Z., Gedney, J., & Vaughan, E. D. (1986). Nipple Differentiation in Fetal Male Rats Treated with an Inhibitor of the Enzyme 5α-Reductase: Definition of a Selective Role for Dihydrotestosterone. <em>Endocrinology</em>, <em>118</em>(1), 132–137. <a href="https://doi.org/10.1210/endo-118-1-132" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1210/endo-118-1-132</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Imperato-McGinley, J., & Gautier, T. (1986). Inherited 5α-reductase deficiency in man. <em>Trends in Genetics</em>, <em>2</em>, 130–133. https://doi.org/https://doi.org/10.1016/0168-9525(86)90202-7</span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Imperato-McGinley, J., Sanchez, R. S., Spencer, J. R., Yee, B., & Darracott Vaughan, E. (1992). Comparison of the Effects of the 5α-Reductase Inhibitor Finasteride and the Antiandrogen Flutamide on Prostate and Genital Differentiation: Dose-Response Studies. <em>Endocrinology</em>, <em>131</em>(3), 1149–1156. <a href="https://doi.org/10.1210/endo.131.3.1324152" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1210/endo.131.3.1324152</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Jarfelt, K., Dalgaard, M., Hass, U., Borch, J., Jacobsen, H., & Ladefoged, O. (2005). Antiandrogenic effects in male rats perinatally exposed to a mixture of di(2-ethylhexyl) phthalate and di(2-ethylhexyl) adipate. <em>Reproductive Toxicology</em>, <em>19</em>(4), 505–515. <a href="https://doi.org/10.1016/j.reprotox.2004.11.005" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1016/j.reprotox.2004.11.005</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Kita, D. H., Meyer, K. B., Venturelli, A. C., Adams, R., Machado, D. L. B., Morais, R. N., Swan, S. H., Gennings, C., & Martino-Andrade, A. J. (2016). Manipulation of pre and postnatal androgen environments and anogenital distance in rats. <em>Toxicology</em>, <em>368–369</em>, 152–161. <a href="https://doi.org/10.1016/j.tox.2016.08.021" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1016/j.tox.2016.08.021</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Kjærstad, M. B., Taxvig, C., Nellemann, C., Vinggaard, A. M., & Andersen, H. R. (2010). Endocrine disrupting effects in vitro of conazole antifungals used as pesticides and pharmaceuticals. <em>Reproductive Toxicology</em>, <em>30</em>(4), 573–582. <a href="https://doi.org/10.1016/j.reprotox.2010.07.009" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1016/j.reprotox.2010.07.009</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Körner, W., Vinggaard, A. M., Térouanne, B., Ma, R., Wieloch, C., Schlumpf, M., Sultan, C., & Soto, A. M. (2004). Interlaboratory comparison of four in vitro assays for assessing androgenic and antiandrogenic activity of environmental chemicals. <em>Environmental Health Perspectives</em>, <em>112</em>(6), 695–702. <a href="https://doi.org/10.1289/ehp.112-1241964" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1289/ehp.112-1241964</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Kratochwil, K. (1977). Development and Loss of Androgen Responsiveness in the Embryonic Rudiment of the Mouse Mammary Gland. <em>DEVELOPMENTAL BIOLOGY</em>, <em>61</em>, 358–365.</span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Kratochwil, K., & Schwartz, P. (1976). Tissue interaction in androgen response of embryonic mammary rudiment of mouse: Identification of target tissue for testosterone (testicular feminization/sexual differentiation/epithelio-mesenchymal interaction). <em>Cell Biology</em>, <em>73</em>(11), 4041–4044.</span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Lee, S.-H., Hong, K. Y., Seo, H., Lee, H.-S., & Park, Y. (2021). Mechanistic insight into human androgen receptor-mediated endocrine-disrupting potentials by a stable bioluminescence resonance energy transfer-based dimerization assay. <em>Chemico-Biological Interactions</em>, <em>349</em>, 109655. <a href="https://doi.org/10.1016/j.cbi.2021.109655" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1016/j.cbi.2021.109655</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Loeffler, I. K., & Peterson, R. E. (1999). Interactive Effects of TCDD and p,p’-DDE on Male Reproductive Tract Development in in Utero and Lactationally Exposed Rats. <em>Toxicology and Applied Pharmacology</em>, <em>154</em>(1), 28–39. <a href="Https://doi.org/10.1006/taap.1998.8572" style="color:#0563c1; text-decoration:underline">Https://doi.org/10.1006/taap.1998.8572</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Lu, S.-Y., Kuo, M.-L., Liao, J.-W., Hwang, J.-S., & Ueng, T.-H. (2006). Antagonistic and Synergistic Effects of Carbendazim and Flutamide Exposures In Utero on Reproductive and Developmental Toxicity in Rats. <em>Journal of Food and Drug Analysis</em>, <em>14</em>(2), 120–132. <a href="https://doi.org/https:/doi.org/10.38212/2224-6614.2491" style="color:#0563c1; text-decoration:underline">https://doi.org/https://doi.org/10.38212/2224-6614.2491</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">MacLean, H. E., Chu, S., Warne, G. L., & Zajac, J. D. (1993). Related individuals with different androgen receptor gene deletions. <em>Journal of Clinical Investigation</em>, <em>91</em>(3), 1123–1128. <a href="https://doi.org/10.1172/JCI116271" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1172/JCI116271</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">MacLeod, D. J., Sharpe, R. M., Welsh, M., Fisken, M., Scott, H. M., Hutchison, G. R., Drake, A. J., & van den Driesche, S. (2010). Androgen action in the masculinization programming window and development of male reproductive organs. <em>International Journal of Andrology</em>, <em>33</em>(2), 279–287. <a href="https://doi.org/10.1111/j.1365-2605.2009.01005.x" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1111/j.1365-2605.2009.01005.x</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Martínez, A. G., Pardo, B., Gámez, R., Mas, R., Noa, M., Marrero, G., Valle, M., García, H., Curveco, D., Mendoza, N., & Goicochea, E. (2011). Effects of in utero exposure to D-004, a lipid extract from roystonea regia fruits, in the male rat: A comparison with finasteride. <em>Journal of Medicinal Food</em>, <em>14</em>(12), 1663–1669. <a href="https://doi.org/10.1089/jmf.2010.0279" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1089/jmf.2010.0279</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Mayer, J. A., Foley, J., de La Cruz, D., Chuong, C. M., & Widelitz, R. (2008). Conversion of the nipple to hair-bearing epithelia by lowering bone morphogenetic protein pathway activity at the dermal-epidermal interface. <em>American Journal of Pathology</em>, <em>173</em>(5), 1339–1348. <a href="https://doi.org/10.2353/ajpath.2008.070920" style="color:#0563c1; text-decoration:underline">https://doi.org/10.2353/ajpath.2008.070920</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">McIntyre, B. S., Barlow, N. J., & Foster, P. M. D. (2001). Androgen-Mediated Development in Male Rat Offspring Exposed to Flutamide in Utero: Permanence and Correlation of Early Postnatal Changes in Anogenital Distance and Nipple Retention with Malformations in Androgen-Dependent Tissues. <em>Toxicological Sciences</em>, <em>62</em>(2), 236–249. <a href="https://doi.org/https:/doi.org/10.1093/toxsci/62.2.236" style="color:#0563c1; text-decoration:underline">https://doi.org/https://doi.org/10.1093/toxsci/62.2.236</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Mcintyre, B. S., Barlow, N. J., & Foster, P. M. D. (2002). Male Rats Exposed to Linuron in Utero Exhibit Permanent Changes in Anogenital Distance, Nipple Retention, and Epididymal Malformations That Result in Subsequent Testicular Atrophy. <em>Toxicological Sciences</em>, <em>65</em>(1), 62–70. <a href="https://doi.org/https:/doi.org/10.1093/toxsci/65.1.62" style="color:#0563c1; text-decoration:underline">https://doi.org/https://doi.org/10.1093/toxsci/65.1.62</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">McIntyre, B. S., Barlow, N. J., Wallace, D. G., Maness, S. C., Gaido, K. W., & Foster, P. M. D. (2000). Effects of in utero exposure to linuron on androgen-dependent reproductive development in the male Crl:CD(SD)BR rat. <em>Toxicology and Applied Pharmacology</em>, <em>167</em>(2), 87–99. <a href="https://doi.org/10.1006/taap.2000.8998" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1006/taap.2000.8998</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Melching-Kollmuss, S., Fussell, K. C., Schneider, S., Buesen, R., Groeters, S., Strauss, V., & van Ravenzwaay, B. (2017). Comparing effect levels of regulatory studies with endpoints derived in targeted anti-androgenic studies: example prochloraz. <em>Archives of Toxicology</em>, <em>91</em>(1), 143–162. <a href="https://doi.org/10.1007/s00204-016-1678-y" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1007/s00204-016-1678-y</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Miyata, K., Yabushita, S., Sukata, T., Sano, M., Yoshino, H., Nakanishi, T., Okuno, Y., & Matsuo, M. (2002). Effects of Perinatal Exposure to Flutamide on Sex Hormones and Androgen-Dependent Organs in F1 Male Rats. <em>The Journal of Toxicological Sciences</em>, <em>27</em>(1), 19–33. <a href="https://doi.org/https:/doi.org/10.2131/jts.27.19" style="color:#0563c1; text-decoration:underline">https://doi.org/https://doi.org/10.2131/jts.27.19</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Moore, R. W., Rudy, T. A., Lin, T.-M., Ko, K., & Peterson, R. E. (2001). Abnormalities of Sexual Development in Male Rats with in Utero and Lactational Exposure to the Antiandrogenic Plasticizer Di(2-ethylhexyl) Phthalate. <em>Environmental Health Perspectives</em>, <em>109</em>(3), 229–237. <a href="http://ehpnet1.niehs.nih.gov/docs/2001/109p229-237moore/abstract.html" style="color:#0563c1; text-decoration:underline">http://ehpnet1.niehs.nih.gov/docs/2001/109p229-237moore/abstract.html</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Mylchreest, E., Sar, M., Cattley, R. C., & Foster, P. M. D. (1999). Disruption of Androgen-Regulated Male Reproductive Development by Di(n-Butyl) Phthalate during Late Gestation in Rats Is Different from Flutamide. <em>Toxicology and Applied Pharmacology</em>, <em>27</em>(1), 81–95. <a href="https://doi.org/https:/doi.org/10.1006/taap.1999.8643" style="color:#0563c1; text-decoration:underline">https://doi.org/https://doi.org/10.1006/taap.1999.8643</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Noriega, N. C., Ostby, J., Lambright, C., Wilson, V. S., & Gray, L. E. (2005). Late gestational exposure to the fungicide prochloraz delays the onset of parturition and causes reproductive malformations in male but not female rat offspring. <em>Biology of Reproduction</em>, <em>72</em>(6), 1324–1335. <a href="https://doi.org/10.1095/biolreprod.104.031385" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1095/biolreprod.104.031385</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">OECD. (2008). Guidance document 43 on mammalian reproductive toxicity testing and assessment. <em>Environment, Health and Safety Publications</em>, <em>16</em>(43).</span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">OECD. (2009). Test Guideline 441: Hershberger Bioassay in Rats: A Short-term Screening Assay for (Anti)Androgenic Properties. <em>OECD Guidelines for the Testing of Chemicals</em>, <em>441</em>.</span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">OECD. (2013). Guidance document supporting OECD test guideline 443 on the extended one-generation reproductive toxicity test. <em>Environment, Health and Safety Publications</em>, <em>10</em>(151).</span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">OECD. (2016a). Test Guideline 421: Reproduction/Developmental Toxicity Screening Test. <em>OECD Guidelines for the Testing of Chemicals</em>, <em>421</em>. <a href="http://www.oecd.org/termsandconditions/" style="color:#0563c1; text-decoration:underline">http://www.oecd.org/termsandconditions/</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">OECD. (2016b). Test Guideline 422: Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test. <em>OECD Guidelines for the Testing of Chemicals</em>, <em>422</em>. <a href="http://www.oecd.org/termsandconditions/" style="color:#0563c1; text-decoration:underline">http://www.oecd.org/termsandconditions/</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">OECD. (2018). Test Guideline 443: Extended one-generation reproductive toxicity study. <em>OECD Guidelines for the Testing of Chemicals</em>, <em>443</em>. <a href="http://www.oecd.org/termsandconditions/" style="color:#0563c1; text-decoration:underline">http://www.oecd.org/termsandconditions/</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">OECD. (2020). Test Guideline 458: Stably Transfected Human Androgen Receptor Transcriptional Activation Assay for Detection of Androgenic Agonist and Antagonist Activity of Chemicals. <em>OECD Guidelines for the Testing of Chemicals</em>, <em>458</em>. <a href="http://www.oecd.org/termsandconditions/" style="color:#0563c1; text-decoration:underline">http://www.oecd.org/termsandconditions/</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Okahashi, N., Sano, M., Miyata, K., Tamano, S., Higuchi, H., Kamita, Y., & Seki, T. (2005). Lack of evidence for endocrine disrupting effects in rats exposed to fenitrothion in utero and from weaning to maturation. <em>Toxicology</em>, <em>206</em>(1), 17–31. <a href="https://doi.org/10.1016/j.tox.2004.04.020" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1016/j.tox.2004.04.020</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Ostby, J., Monosson, E., Kelce, W. R., & Earl Gray, L. J. (1999). Environmental antiandrogens: low doses of the fungicide vinclozolin alter sexual differentiation of the male rat. <em>Toxicology and Industrial Health</em>, <em>15</em>, 48–64. <a href="https://doi.org/https:/doi.org/10.1177/074823379901500106" style="color:#0563c1; text-decoration:underline">https://doi.org/https://doi.org/10.1177/074823379901500106</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Panagiotou, E. M., Draskau, M. K., Li, T., Hirschberg, A., Svingen, T., & Damdimopoulou, P. (2022). AOP key event relationship report: Linking decreased androgen receptor activation with decreased granulosa cell proliferation of gonadotropin-independent follicles. Reproductive Toxicology, 112, 136-147. <a href="https://doi.org/10.1016/j.reprotox.2022.07.004" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1016/j.reprotox.2022.07.004</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Pedersen, E. B., Christiansen, S., & Svingen, T. (2022). AOP key event relationship report: Linking androgen receptor antagonism with nipple retention. <em>Current Research in Toxicology</em>, 3, 100085. https://doi.org/10.1016/j.crtox.2022.100085 </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Rana, K., Davey, R., & Zajac, J. (2014). Human androgen deficiency: insights gained from androgen receptor knockout mouse models. <em>Asian Journal of Andrology</em>, <em>16</em>(2), 169. <a href="https://doi.org/10.4103/1008-682X.122590" style="color:#0563c1; text-decoration:underline"><span style="color:#0563c1">https://doi.org/10.4103/1008-682X.122590</span></a></span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Rider CV, Furr JR, Wilson VS, Gray LE Jr (2010). Cumulative effects of in utero administration of mixtures of reproductive toxicants that disrupt common target tissues via diverse mechanisms of toxicity. <em>Int J Androl</em>. 33(2), 443-62. <a href="https://doi.org/10.1111/j.1365-2605.2009.01049.x" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1111/j.1365-2605.2009.01049.x</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Saillenfait, A. M., Sabaté, J. P., & Gallissot, F. (2008). Diisobutyl phthalate impairs the androgen-dependent reproductive development of the male rat. <em>Reproductive Toxicology</em>, <em>26</em>(2), 107–115. <a href="https://doi.org/10.1016/j.reprotox.2008.07.006" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1016/j.reprotox.2008.07.006</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Saillenfait, A. M., Sabaté, J. P., & Gallissot, F. (2009). Effects of in utero exposure to di-n-hexyl phthalate on the reproductive development of the male rat. <em>Reproductive Toxicology</em>, <em>28</em>(4), 468–476. <a href="https://doi.org/10.1016/j.reprotox.2009.06.013" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1016/j.reprotox.2009.06.013</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Satoh, K., Ohyama, K., Aoki, N., Iida, M., & Nagai, F. (2004). Study on anti-androgenic effects of bisphenol a diglycidyl ether (BADGE), bisphenol F diglycidyl ether (BFDGE) and their derivatives using cells stably transfected with human androgen receptor, AR-EcoScreen. <em>Food and Chemical Toxicology</em>, <em>42</em>(6), 983–993. <a href="https://doi.org/10.1016/j.fct.2004.02.011" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1016/j.fct.2004.02.011</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Schneider, S., Kaufmann, W., Strauss, V., & van Ravenzwaay, B. (2011). Vinclozolin: A feasibility and sensitivity study of the ILSI-HESI F1-extended one-generation rat reproduction protocol. <em>Regulatory Toxicology and Pharmacology</em>, <em>59</em>(1), 91–100. <a href="https://doi.org/10.1016/j.yrtph.2010.09.010" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1016/j.yrtph.2010.09.010</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Schreiber, E., Garcia, T., González, N., Esplugas, R., Sharma, R. P., Torrente, M., Kumar, V., Bovee, T., Katsanou, E. S., Machera, K., Domingo, J. L., & Gómez, M. (2020). Maternal exposure to mixtures of dienestrol, linuron and flutamide. Part I: Feminization effects on male rat offspring. <em>Food and Chemical Toxicology</em>, <em>139</em>(1), 1–13. <a href="https://doi.org/10.1016/j.fct.2020.111256" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1016/j.fct.2020.111256</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Schwartz, C. L., Christiansen, S., Hass, U., Ramhøj, L., Axelstad, M., Löbl, N. M., & Svingen, T. (2021). On the Use and Interpretation of Areola/Nipple Retention as a Biomarker for Anti-androgenic Effects in Rat Toxicity Studies. <em>Frontiers in Toxicology</em>, <em>3</em>. <a href="https://doi.org/10.3389/ftox.2021.730752" style="color:#0563c1; text-decoration:underline">https://doi.org/10.3389/ftox.2021.730752</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Schwartz, C. L., Christiansen, S., Vinggaard, A. M., Axelstad, M., Hass, U., & Svingen, T. (2019). Anogenital distance as a toxicological or clinical marker for fetal androgen action and risk for reproductive disorders. <em>Archives of Toxicology</em>, <em>93</em>(2), 253–272. <a href="https://doi.org/10.1007/s00204-018-2350-5" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1007/s00204-018-2350-5</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Sonneveld, E., Jansen, H. J., Riteco, J. A. C., Brouwer, A., & van der Burg, B. (2005). Development of Androgen- and Estrogen-Responsive Bioassays, Members of a Panel of Human Cell Line-Based Highly Selective Steroid-Responsive Bioassays. <em>Toxicological Sciences</em>, <em>83</em>(1), 136–148. <a href="https://doi.org/10.1093/toxsci/kfi005" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1093/toxsci/kfi005</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Svingen, T., Villeneuve, D. L., Knapen, D., Panagiotou, E. M., Draskau, M. K., Damdimopoulou, P., & O’Brien, J. M. (2021). A Pragmatic Approach to Adverse Outcome Pathway Development and Evaluation. <em>Toxicological Sciences</em>, <em>184</em>(2), 183–190. https://doi.org/10.1093/toxsci/kfab113</span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Taxvig, C., Hass, U., Axelstad, M., Dalgaard, M., Boberg, J., Andeasen, H. R., & Vinggaard, A. M. (2007). Endocrine-disrupting activities In Vivo of the fungicides tebuconazole and epoxiconazole. <em>Toxicological Sciences</em>, <em>100</em>(2), 464–473. <a href="https://doi.org/10.1093/toxsci/kfm227" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1093/toxsci/kfm227</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Turner, K. J., Barlow, N. J., Struve, M. F., Wallace, D. G., Gaido, K. W., Dorman, D. C., & Foster, P. M. D. (2002). Effects of in Utero Exposure to the Organophosphate Insecticide Fenitrothion on Androgen-Dependent Reproductive Development in the Crl:CD(SD)BR Rat. <em>Toxicological Sciences</em>, <em>68</em>(1), 174–183. <a href="https://doi.org/https:/doi.org/10.1093/toxsci/68.1.174" style="color:#0563c1; text-decoration:underline">https://doi.org/https://doi.org/10.1093/toxsci/68.1.174</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">van der Burg, B., Winter, R., Man, H., Vangenechten, C., Berckmans, P., Weimer, M., Witters, H., & van der Linden, S. (2010). Optimization and prevalidation of the in vitro AR CALUX method to test androgenic and antiandrogenic activity of compounds. <em>Reproductive Toxicology</em>, <em>30</em>(1), 18–24. <a href="https://doi.org/10.1016/j.reprotox.2010.04.012" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1016/j.reprotox.2010.04.012</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Vinggaard, A. M., Christiansen, S., Laier, P., Poulsen, M. E., Breinholt, V., Jarfelt, K., Jacobsen, H., Dalgaard, M., Nellemann, C., & Hass, U. (2005). Perinatal exposure to the fungicide prochloraz feminizes the male rat offspring. <em>Toxicological Sciences</em>, <em>85</em>(2), 886–897. <a href="https://doi.org/10.1093/toxsci/kfi150" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1093/toxsci/kfi150</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Vinggaard, A. M., Hass, U., Dalgaard, M., Andersen, H. R., Bonefeld-Jørgensen, E., Christiansen, S., Laier, P., Poulsen, M. E., McLachlan, J., Main, K. M., Søeborg, T., & Foster, P. (2006). Prochloraz: An imidazole fungicide with multiple mechanisms of action. <em>International Journal of Andrology</em>, <em>29</em>(1), 186–192. <a href="https://doi.org/10.1111/j.1365-2605.2005.00604.x" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1111/j.1365-2605.2005.00604.x</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Vinggaard, A. M., Niemelä, J., Wedebye, E. B., & Jensen, G. E. (2008). Screening of 397 Chemicals and Development of a Quantitative Structure−Activity Relationship Model for Androgen Receptor Antagonism. <em>Chemical Research in Toxicology</em>, <em>21</em>(4), 813–823. <a href="https://doi.org/10.1021/tx7002382" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1021/tx7002382</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Walters, K. A., Simanainen, U., & Handelsman, D. J. (2010). Molecular insights into androgen actions in male and female reproductive function from androgen receptor knockout models. <em>Human Reproduction Update</em>, <em>16</em>(5), 543–558. <a href="https://doi.org/10.1093/humupd/dmq003" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1093/humupd/dmq003</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Welsh, M., Saunders, P. T. K., Fisken, M., Scott, H. M., Hutchison, G. R., Smith, L. B., & Sharpe, R. M. (2008). Identification in rats of a programming window for reproductive tract masculinization, disruption of which leads to hypospadias and cryptorchidism. <em>Journal of Clinical Investigation</em>, <em>118</em>(4), 1479–1490. <a href="https://doi.org/10.1172/JCI34241" style="color:#0563c1; text-decoration:underline"><span style="color:#0563c1">https://doi.org/10.1172/JCI34241</span></a></span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Wilson, V.S., Bobseine, K., Earl Gray Jr, L. (2004), Development and Characterization of a Cell Line That Stably Expresses an Estrogen-Responsive Luciferase Reporter for the Detection of Estrogen Receptor Agonist and Antagonists. <em>Toxicological Sciences</em>, 81, 69-77. <a href="https://doi.org/10.1093/toxsci/kfh180" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1093/toxsci/kfh180</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Wolf, C. J., LeBlanc, G. A., & Gray, L. E. (2004). Interactive effects of vinclozolin and testosterone propionate on pregnancy and sexual differentiation of the male and female SD rat. <em>Toxicological Sciences</em>, <em>78</em>(1), 135–143. <a href="https://doi.org/10.1093/toxsci/kfh018" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1093/toxsci/kfh018</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Wolf, Lambright, C., Mann, P., Price, M., Cooper, R. L., Ostby, J., & Earl Gray, L. J. (1999). Administration of potentially antiandrogenic pesticides (procymidone, linuron, iprodione, chlozolinate, p,p-DDE, and ketoconazole) and toxic substances (dibutyl-and diethylhexyl phthalate, PCB 169, and ethane dimethane sulphonate) during sexual differentiation produces diverse profiles of reproductive malformations in the male rat. <em>Toxicology and Industrial Health</em>, <em>15</em>(2), 94–118. <a href="https://doi.org/https:/doi.org/10.1177/074823379901500109" style="color:#0563c1; text-decoration:underline">https://doi.org/https://doi.org/10.1177/074823379901500109</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Wolf, Leblanc, G. A., Ostby, J. S., Gray, L. E., & Branch, E. (2000). Characterization of the Period of Sensitivity of Fetal Male Sexual Development to Vinclozolin. <em>Toxicological Sciences</em>, <em>55</em>(1), 152–161. <a href="https://doi.org/https:/doi.org/10.1093/toxsci/55.1.152" style="color:#0563c1; text-decoration:underline">https://doi.org/https://doi.org/10.1093/toxsci/55.1.152</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Yamasaki, K., Okuda, H., Takeuchi, T., & Minobe, Y. (2009). Effects of in utero through lactational exposure to dicyclohexyl phthalate and p,p′-DDE in Sprague-Dawley rats. <em>Toxicology Letters</em>, <em>189</em>(1), 14–20. <a href="https://doi.org/10.1016/j.toxlet.2009.04.023" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1016/j.toxlet.2009.04.023</a> </span></span></p>
<p style="margin-left:38px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">You, P.-D., Casanova, L., Archibeque-Engle, M., Sar, S., Fan, M., Heck, L.-Q. A., & D’a, H. (1998). Impaired Male Sexual Development in Perinatal Sprague-Dawley and Long-Evans Hooded Rats Exposed in Utero and Lactationally to p,p’-DDE. <em>Toxicol. Sci</em>, <em>45</em>, 162–173. </span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"><a href="https://doi.org/10.1093/toxsci/45.2.162" style="color:#0563c1; text-decoration:underline">https://doi.org/10.1093/toxsci/45.2.162</a></span></span></p>
<p> </p>
2020-05-11T10:24:312023-01-11T06:22:468582eaef-c27c-4a3d-b719-a1154db2f653d68bce64-3ad8-4410-9957-fa2216eddbb7<p style="text-align:justify"><span style="font-size:12pt">The androgen receptor (AR) is a ligand-dependent nuclear transcription factor that upon activation translocates to the nucleus, dimerizes, and binds androgen response elements (AREs) to modulate transcription of target genes <span style="color:black">(Lamont and Tindall, 2010, Roy et al. 2001)</span>. Decreased activation of the AR affects its transcription factor activity, therefore leading to altered AR-target gene expression. This KER refers to decreased AR activation and altered gene expression occurring in complex systems, such as <em>in vivo</em> and the specific effect on transcription of AR target genes will depend on species, life stage, tissue, cell type etc. </span></p>
<p style="text-align:justify"><span style="font-size:12pt">The biological plausibility for this KER is considered high</span></p>
<p style="text-align:justify"><span style="font-size:12pt">The AR is a ligand-activated transcription factor part of the steroid hormone nuclear receptor family. Non-activated AR is found in the cytoplasm as a multiprotein complex with heat-shock proteins, immunophilins and, other chaperones <span style="color:black">(Roy et al. 2001)</span>. Upon activation through ligand binding, the AR dissociates from the protein complex, translocates to the nucleus and homodimerizes. Facilitated by co-regulators, AR can bind to DNA regions containing AREs and initiate transcription of target genes, that thus will be different in e.g. different tissues, life-stages, species etc. </span></p>
<p style="text-align:justify"><span style="font-size:12pt">Through mapping of AREs and ChIP sequencing studies, several AR target genes have been identified, mainly studied in prostate cells <span style="color:black">(Jin, Kim, and Yu 2013)</span>. Different co-regulators and ligands lead to altered expression of different sets of genes <span style="color:black">(Jin et al. 2013; Kanno et al. 2022)</span>. Alternative splicing of the AR can lead to different AR variants that also affects which genes are transcribed <span style="color:black">(Jin et al. 2013)</span>.</span></p>
<p style="text-align:justify"><span style="font-size:12pt">Apart from this canonical signaling pathway, the AR can suppress gene expression, indirectly regulate miRNA transcription, and have non-genomic effects by rapid activation of second messenger pathways in either presence or absence of a ligand <span style="color:black">(Jin et al. 2013)</span>.</span></p>
<p style="text-align:justify"><span style="font-size:12pt">The empirical evidence for this KER is considered high</span></p>
<p style="text-align:justify"><span style="font-size:12pt">In humans, altered gene expression profiling in individuals with androgen insensitivity syndrome (AIS) can provide supporting empirical evidence <span style="color:black">(Holterhus et al. 2003; Peng et al. 2021)</span>. In rodent AR knockout (KO) models, gene expression profiling studies and gene-targeted approaches have provided information on differentially expressed genes in several organ systems including male and female reproductive, endocrine, muscular, cardiovascular and nervous systems <span style="color:black">(Denolet et al. 2006; Fan et al. 2005; Holterhus et al. 2003; Ikeda et al. 2005; Karlsson et al. 2016; MacLean et al. 2008; Rana et al. 2011; Russell et al. 2012; Shiina et al. 2006; Wang et al. 2006; Welsh et al. 2012; Willems et al. 2010; Yu et al. 2008, 2012; Zhang et al. 2006; Zhou et al. 2011)</span>.</span></p>
<p style="text-align:justify"><span style="font-size:12pt">Exposure to known antiandrogens has been shown to alter transcriptional profiles, for example of neonatal pig ovaries <span style="color:black">(Knapczyk-Stwora et al. 2019)</span>. </span></p>
<p style="text-align:justify"><span style="font-size:12pt">Dose concordance has also been observed for instance in zebrafish embryos; a dose of 50 µg/L of the AR antagonist flutamide resulted in 674 differentially expressed genes at 96 h post fertilization whereas 500 µg/L flutamide resulted in 2871 differentially expressed genes (Ayobahan et al., 2023). </span></p>
<p style="text-align:justify"><span style="font-size:12pt">AR action has been reported to occur also without ligand binding. However, not much is known about the extent and biological implications of such non-canonical, ligand-independent AR activation <span style="color:black">(Bennesch and Picard 2015)</span>.</span></p>
HighMixedHighDuring development and at adulthoodHigh<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Ayobahan, S. U., Alvincz, J., Reinwald, H., Strompen, J., Salinas, G., Schäfers, C., et al. (2023). Comprehensive identification of gene expression fingerprints and biomarkers of sexual endocrine disruption in zebrafish embryo. Ecotoxicol. Environ. Saf. 250, 114514. doi:10.1016/J.ECOENV.2023.114514.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Bennesch, Marcela A., and Didier Picard. 2015. “Minireview: Tipping the Balance: Ligand-Independent Activation of Steroid Receptors.” <em>Molecular Endocrinology</em> 29(3):349–63.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Chamberlain, Nancy L., Erika D. Driverand, and Roger L. Miesfeldi. 1994. <em>The Length and Location of CAG Trinucleotide Repeats in the Androgen Receptor N-Terminal Domain Affect Transactivation Function</em>. Vol. 22.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Denolet, Evi, Karel De Gendt, Joke Allemeersch, Kristof Engelen, Kathleen Marchal, Paul Van Hummelen, Karen A. L. Tan, Richard M. Sharpe, Philippa T. K. Saunders, Johannes V. Swinnen, and Guido Verhoeven. 2006. “The Effect of a Sertoli Cell-Selective Knockout of the Androgen Receptor on Testicular Gene Expression in Prepubertal Mice.” <em>Molecular Endocrinology</em> 20(2):321–34. doi: 10.1210/me.2005-0113.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Fan, Wuqiang, Toshihiko Yanase, Masatoshi Nomura, Taijiro Okabe, Kiminobu Goto, Takashi Sato, Hirotaka Kawano, Shigeaki Kato, and Hajime Nawata. 2005. <em>Androgen Receptor Null Male Mice Develop Late-Onset Obesity Caused by Decreased Energy Expenditure and Lipolytic Activity but Show Normal Insulin Sensitivity With High Adiponectin Secretion</em>. Vol. 54.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Holterhus, Paul-Martin, Olaf Hiort, Janos Demeter, Patrick O. Brown, and James D. Brooks. 2003. <em>Differential Gene-Expression Patterns in Genital Fibroblasts of Normal Males and 46,XY Females with Androgen Insensitivity Syndrome: Evidence for Early Programming Involving the Androgen Receptor</em>. Vol. 4.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Ikeda, Yasumasa, Ken Ichi Aihara, Takashi Sato, Masashi Akaike, Masanori Yoshizumi, Yuki Suzaki, Yuki Izawa, Mitsunori Fujimura, Shunji Hashizume, Midori Kato, Shusuke Yagi, Toshiaki Tamaki, Hirotaka Kawano, Takahiro Matsumoto, Hiroyuki Azuma, Shigeaki Kato, and Toshio Matsumoto. 2005. “Androgen Receptor Gene Knockout Male Mice Exhibit Impaired Cardiac Growth and Exacerbation of Angiotensin II-Induced Cardiac Fibrosis.” <em>Journal of Biological Chemistry</em> 280(33):29661–66. doi: 10.1074/jbc.M411694200.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Jin, Hong Jian, Jung Kim, and Jindan Yu. 2013. “Androgen Receptor Genomic Regulation.” <em>Translational Andrology and Urology</em> 2(3):158–77.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Kang, Zhigang, Asta Pirskanen, Olli A. Jänne, and Jorma J. Palvimo. 2002. “Involvement of Proteasome in the Dynamic Assembly of the Androgen Receptor Transcription Complex.” <em>Journal of Biological Chemistry</em> 277(50):48366–71. doi: 10.1074/jbc.M209074200.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Kanno, Yuichiro, Nao Saito, Ryota Saito, Tomohiro Kosuge, Ryota Shizu, Tomofumi Yatsu, Takuomi Hosaka, Kiyomitsu Nemoto, Keisuke Kato, and Kouichi Yoshinari. 2022. “Differential DNA-Binding and Cofactor Recruitment Are Possible Determinants of the Synthetic Steroid YK11-Dependent Gene Expression by Androgen Receptor in Breast Cancer MDA-MB 453 Cells.” <em>Experimental Cell Research</em> 419(2). doi: 10.1016/j.yexcr.2022.113333.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Karlsson, Sara A., Erik Studer, Petronella Kettunen, and Lars Westberg. 2016. “Neural Androgen Receptors Modulate Gene Expression and Social Recognition but Not Social Investigation.” <em>Frontiers in Behavioral Neuroscience</em> 10(MAR). doi: 10.3389/fnbeh.2016.00041.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Knapczyk-Stwora, Katarzyna, Anna Nynca, Renata E. Ciereszko, Lukasz Paukszto, Jan P. Jastrzebski, Elzbieta Czaja, Patrycja Witek, Marek Koziorowski, and Maria Slomczynska. 2019. “Flutamide-Induced Alterations in Transcriptional Profiling of Neonatal Porcine Ovaries.” <em>Journal of Animal Science and Biotechnology</em> 10(1):1–15. doi: 10.1186/s40104-019-0340-y.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Lamont, K. R., and Tindall, D. J. (2010). Androgen Regulation of Gene Expression. Adv. Cancer Res. 107, 137–162. doi:10.1016/S0065-230X(10)07005-3.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">MacLean, Helen E., W. S. Maria Chiu, Amanda J. Notini, Anna-Maree Axell, Rachel A. Davey, Julie F. McManus, Cathy Ma, David R. Plant, Gordon S. Lynch, and Jeffrey D. Zajac. 2008. “ Impaired Skeletal Muscle Development and Function in Male, but Not Female, Genomic Androgen Receptor Knockout Mice .” <em>The FASEB Journal</em> 22(8):2676–89. doi: 10.1096/fj.08-105726.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Maiuri, Paolo, Anna Knezevich, Alex De Marco, Davide Mazza, Anna Kula, Jim G. McNally, and Alessandro Marcello. 2011. “Fast Transcription Rates of RNA Polymerase II in Human Cells.” <em>EMBO Reports</em> 12(12):1280–85. doi: 10.1038/embor.2011.196.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Mora, Gloria R., and Virendra B. Mahesh. 1999. <em>Autoregulation of the Androgen Receptor at the Translational Level: Testosterone Induces Accumulation of Androgen Receptor MRNA in the Rat Ventral Prostate Polyribosomes</em>.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Peng, Yajie, Hui Zhu, Bing Han, Yue Xu, Xuemeng Liu, Huaidong Song, and Jie Qiao. 2021. “Identification of Potential Genes in Pathogenesis and Diagnostic Value Analysis of Partial Androgen Insensitivity Syndrome Using Bioinformatics Analysis.” <em>Frontiers in Endocrinology</em> 12. doi: 10.3389/fendo.2021.731107.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Rana, Kesha, Barbara C. Fam, Michele V Clarke, Tammy P. S. Pang, Jeffrey D. Zajac, and Helen E. Maclean. 2011. “Increased Adiposity in DNA Binding-Dependent Androgen Receptor Knockout Male Mice Associated with Decreased Voluntary Activity and Not Insulin Resistance.” <em>Am J Physiol Endocrinol Me-Tab</em> 301:767–78. doi: 10.1152/ajpendo.00584.2010.-In.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Roy, Arun K., Rakesh K. Tyagi, Chung S. Song, Yan Lavrovsky, Soon C. Ahn, Tae Sung Oh, and Bandana Chatterjee. 2001. “Androgen Receptor: Structural Domains and Functional Dynamics after Ligand-Receptor Interaction.” Pp. 44–57 in <em>Annals of the New York Academy of Sciences</em>. Vol. 949. New York Academy of Sciences.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Russell, Patricia K., Michele V. Clarke, Jarrod P. Skinner, Tammy P. S. Pang, Jeffrey D. Zajac, and Rachel A. Davey. 2012. “Identification of Gene Pathways Altered by Deletion of the Androgen Receptor Specifically in Mineralizing Osteoblasts and Osteocytes in Mice.” <em>Journal of Molecular Endocrinology</em> 49(1):1–10. doi: 10.1530/JME-12-0014.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Shiina, Hiroko, Takahiro Matsumoto, Takashi Sato, Katsuhide Igarashi, Junko Miyamoto, Sayuri Takemasa, Matomo Sakari, Ichiro Takada, Takashi Nakamura, Daniel Metzger, Pierre Chambon, Jun Kanno, Hiroyuki Yoshikawa, and Shigeaki Kato. 2006. <em>Premature Ovarian Failure in Androgen Receptor-Deficient Mice</em>. Vol. 103.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Supakar, P. C., C. S. Song, M. H. Jung, M. A. Slomczynska, J. M. Kim, R. L. Vellanoweth, B. Chatterjee, and A. K. Roy. 1993. “A Novel Regulatory Element Associated with Age-Dependent Expression of the Rat Androgen Receptor Gene.” <em>Journal of Biological Chemistry</em> 268(35):26400–408. doi: 10.1016/s0021-9258(19)74328-2.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Tut, Thein G., Farid J. Ghadessy, M. A. Trifiro, L. Pinsky, and E. L. Yong. 1997. <em>Long Polyglutamine Tracts in the Androgen Receptor Are Associated with Reduced Trans-Activation, Impaired Sperm Production, and Male Infertility*</em>. Vol. 82.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Wang, Ruey Sheng, Shuyuan Yeh, Lu Min Chen, Hung Yun Lin, Caixia Zhang, Jing Ni, Cheng Chia Wu, P. Anthony Di Sant’Agnese, Karen L. DeMesy-Bentley, Chii Ruey Tzeng, and Chawnshang Chang. 2006. “Androgen Receptor in Sertoli Cell Is Essential for Germ Cell Nursery and Junctional Complex Formation in Mouse Testes.” <em>Endocrinology</em> 147(12):5624–33. doi: 10.1210/en.2006-0138.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Welsh, M., L. Moffat, K. Belling, L. R. de França, T. M. Segatelli, P. T. K. Saunders, R. M. Sharpe, and L. B. Smith. 2012. “Androgen Receptor Signalling in Peritubular Myoid Cells Is Essential for Normal Differentiation and Function of Adult Leydig Cells.” <em>International Journal of Andrology</em> 35(1):25–40. doi: 10.1111/j.1365-2605.2011.01150.x.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Willems, Ariane, Sergio R. Batlouni, Arantza Esnal, Johannes V. Swinnen, Philippa T. K. Saunders, Richard M. Sharpe, Luiz R. França, Karel de Gendt, and Guido Verhoeven. 2010. “Selective Ablation of the Androgen Receptor in Mouse Sertoli Cells Affects Sertoli Cell Maturation, Barrier Formation and Cytoskeletal Development.” <em>PLoS ONE</em> 5(11). doi: 10.1371/journal.pone.0014168.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Wu, D. I., Grace Lin, and Andrea C. Gore. 2009. “Age-Related Changes in Hypothalamic Androgen Receptor and Estrogen Receptor in Male Rats.” <em>The Journal of Comparative Neurology</em> 512:688–701. doi: 10.1002/cne.21925.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Yu, I. Chen, Hung Yun Lin, Ning Chun Liu, Ruey Shen Wang, Janet D. Sparks, Shuyuan Yeh, and Chawnshang Chang. 2008. “Hyperleptinemia without Obesity in Male Mice Lacking Androgen Receptor in Adipose Tissue.” <em>Endocrinology</em> 149(5):2361–68. doi: 10.1210/en.2007-0516.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Yu, Shengqiang, Chiuan Ren Yeh, Yuanjie Niu, Hong Chiang Chang, Yu Chieh Tsai, Harold L. Moses, Chih Rong Shyr, Chawnshang Chang, and Shuyuan Yeh. 2012. “Altered Prostate Epithelial Development in Mice Lacking the Androgen Receptor in Stromal Fibroblasts.” <em>Prostate</em> 72(4):437–49. doi: 10.1002/pros.21445.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Zhang, Caixia, Shuyuan Yeh, Yen-Ta Chen, Cheng-Chia Wu, Kuang-Hsiang Chuang, Hung-Yun Lin, Ruey-Sheng Wang, Yu-Jia Chang, Chamindrani Mendis-Handagama, Liquan Hu, Henry Lardy, Chawnshang Chang, and † † George. 2006. <em>Oligozoospermia with Normal Fertility in Male Mice Lacking the Androgen Receptor in Testis Peritubular Myoid Cells</em>.</span></span></p>
<p><span style="font-size:12pt"><span style="font-family:Calibri,sans-serif">Zhou, Wei, Gensheng Wang, Christopher L. Small, Zhilin Liu, Connie C. Weng, Lizhong Yang, Michael D. Griswold, and Marvin L. Meistrich. 2011. “Erratum: Gene Expression Alterations by Conditional Knockout of Androgen Receptor in Adult Sertoli Cells of Utp14bjsd/Jsd (Jsd) Mice (Biology of Reproduction (2010) 83, (759-766) DOI: 10.1095/Biolreprod.110.085472).” <em>Biology of Reproduction</em> 84(2):400–408.</span></span></p>
2020-05-11T06:50:062023-10-19T08:42:39d68bce64-3ad8-4410-9957-fa2216eddbb76efacf59-b918-4fa7-9464-a0b0b83609302020-05-11T10:24:112020-05-11T10:24:11Androgen receptor (AR) antagonism leading to nipple retention (NR) in male (mammalian) offspringAR antagonism leading to NR<p>Sofie Christiansen; National Food Institute, Technical University of Denmark, Kongens Lyngby, 2800 Denmark</p>
<p>Louise Ramhøj; National Food Institute, Technical University of Denmark, Kongens Lyngby, 2800 Denmark</p>
<p>Terje Svingen; National Food Institute, Technical University of Denmark, Kongens Lyngby, 2800 Denmark</p>
Under development: Not open for comment. Do not citeUnder DevelopmentIncluded in OECD Work Plan1.108<p>This AOP links Androgen receptor antagonism during fetal life with nipple retention (NR) in male rat offspring. NR, measured around 2 weeks post partum, is a marker for feminization of male rat fetuses and is associated with general feminization of male offspring. Although NR is not a directly applicable measure in humans (male humans normally retain two nipples), it is nevertheless considered an ‘adverse outcome’ in OECD test guidelines; NR measurements are mandatory in specific tests for developmental and reproductive toxicity in chemical risk assessment (TG 443, TG 421/422, TG 414).</p>
<p>The AR is a nuclear receptor involved in the transcriptional regulation of various target genes during development and adulthood across species. Its main ligand is testosterone and dihydrotestosterone (DHT). Under normal physiological conditions, testosterone produced mainly by the testicles, is converted in peripheral tissues by 5α-reductase into DHT, which in turn binds AR and activates downstream target genes. AR signaling is necessary for normal masculinization of the developing fetus, and AR action in male rats signals the nipple anlagen to regress, leaving males with no nipples.</p>
<div>The key events in this pathway is antagonism of the AR in target cells of the nipple anlagen, which leads to inactivation of the AR and failure to suppress development of the nipples. In this instance, the local levels of testosterone or DHT may be normal, but prevented from binding the AR.</div>
<p>A large number of drugs and chemicals have been shown to antagonise the AR using various AR reporter gene assays. The AR is specifically targeted in AR-sensitive cancers, for example the use of the anti-androgenic drug flutamide in treating prostate cancer (<a href="#_ENREF_1" title="Alapi, 2006 #262">Alapi & Fischer, 2006</a>). Flutamide has also been used in several rodent in vivo studies showing anti-androgenic effects (feminization of male offspring) evident by e.g. short anogenital distance (AGD) in males (<a href="#_ENREF_4" title="Foster, 2005 #53">Foster & Harris, 2005</a>; <a href="#_ENREF_5" title="Hass, 2007 #76">Hass et al, 2007</a>; <a href="#_ENREF_8" title="Kita, 2016 #34">Kita et al, 2016</a>). QSAR models can predict AR antagonism for a wide range of chemicals, many of which have shown in vitro antagonistic potential (<a href="#_ENREF_17" title="Vinggaard, 2008 #263">Vinggaard et al, 2008</a>).</p>
<p><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">NR is recognized by the OECD as a relevant measure for anti-androgenic effects and is mandatory in the test guidelines Extended One Generation Reproductive Toxicity Study, TG 443 <span style="color:black">(OECD, 2018) </span>and the two screening studies for reproductive toxicity, TGs 421/422 <span style="color:black">(OECD, 2016a, 2016b)</span>. The endpoint is also described in the guidance documents 43 <span style="color:black">(OECD, 2008)</span> and 151 <span style="color:black">(OECD, 2013)</span>. Furthermore, NR data can be used in chemical risk assessment for setting the No Observed Adverse Effect Level (NOAEL) as stated in the OECD guidance document 151 <span style="color:black">(OECD, 2013)</span>: “<em>A statistically significant change in nipple retention should be evaluated similarly to an effect on AGD as both endpoints indicate an adverse effect of exposure and should be considered in setting a NOAEL</em>”.</span></span></p>
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