13311-84-7MKXKFYHWDHIYRV-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
DTXSID7032004131983-72-7PPDBOQMNKNNODG-UHFFFAOYNA-NPPDBOQMNKNNODG-UHFFFAOYSA-N
Triticonazole5-[(4-Chlorophenyl)methylene]-2,2-dimethyl-1-(1H-1,2,4-triazol-1-ylmethyl)cyclopentanol
DTXSID003265585509-19-9FQKUGOMFVDPBIZ-UHFFFAOYSA-NFQKUGOMFVDPBIZ-UHFFFAOYSA-N
FlusilazoleNuStar
DTXSID3024235133855-98-8ZMYFCFLJBGAQRS-UHFFFAOYNA-NZMYFCFLJBGAQRS-UHFFFAOYSA-N
EpoxiconazoleDTXSID104037267747-09-5TVLSRXXIMLFWEO-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
DTXSID8024280107534-96-3PXMNMQRDXWABCY-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
DTXSID9032113427-51-0UWFYSQMTEOIJJG-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
DTXSID502036650471-44-8FSCWZHGZWWDELK-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
DTXSID402236190357-06-5LKJPYSCBVHEWIU-UHFFFAOYSA-NLKJPYSCBVHEWIU-UHFFFAOYSA-N
BicalutamideCasodex
CDX
Propanamide, N-[4-cyano-3-(trifluoromethyl)phenyl]-3-[(4-fluorophenyl)sulfonyl]-2-hydroxy-2-methyl-
DTXSID2022678PR:000004191androgen receptorGO:0010468regulation of gene expressionVT:1000294egg quantity2decreasedFlutamide2016-11-29T18:42:272016-11-29T18:42:27Mercaptobenzole2016-11-29T18:42:262016-11-29T18:42:26Triticonazole2020-05-16T11:02:072020-05-16T11:09:42Flusilazole2020-05-16T11:15:342020-05-16T11:15:34Epoxiconazole2020-05-16T11:35:442020-05-16T11:35:44Prochloraz2016-11-29T18:42:222016-11-29T18:42:22Propiconazole2017-05-17T13:18:072017-05-17T13:18:07Tebuconazole2017-05-17T13:17:142017-05-17T13:17:14Cyproterone acetate2020-05-17T10:13:282020-05-17T10:13:28Vinclozolin2020-05-14T11:28:312020-05-14T11:28:31Bicalutamide2020-08-07T06:55:532020-08-07T06:55:53Stressor:286 Tebuconazole2020-08-07T07:00:532020-08-07T07:00:53Vinclozalin2016-11-29T18:42:272016-11-29T18:42:27WCS_9606human10090mouse10116ratWikiUser_25human and other cells in cultureWikiUser_14MonkeyWikiUser_24Pig9913cowWCS_7955zebrafishAntagonism, Androgen receptorAntagonism, Androgen receptorMolecular<p><u>The androgen receptor (AR) and its function</u></p>
<p>Development of the male reproductive system and secondary male characteristics is dependent on androgens (foremost testosterone (T) and dihydrotestosterone (DHT). T and the more biologically active DHT act by binding to 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>), with human AR mutations and mouse knock-out models having established its pivotal role in masculinization and spermatogenesis (<a href="#_ENREF_9" title="Walters, 2010 #254">Walters et al, 2010</a>). The AR is a ligand-activated transcription factor belonging to the steroid hormone nuclear receptor family (<a href="#_ENREF_1" title="Davey, 2016 #250">Davey & Grossmann, 2016</a>). 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. Apart from the essential role AR plays for 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 function through non-genomic modalities, for instance rapid change in cell function by ion transport changes (<a href="#_ENREF_3" title="Heinlein, 2002 #256">Heinlein & Chang, 2002</a>). However, with regard to this specific KE the canonical signaling pathway is what is referred to.</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 is freely available for 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="#_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"><span style="font-family:"Calibri",sans-serif">Recently developed AR dimerization assay may soon be included in TGs for its improved ability to measure potential stressor-mediated dimerization/activation </span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">(</span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"><a href="#_ENREF_11" title="Lee, 2021 #288">Lee et al, 2021</a></span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">)</span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">.</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>).</p>
<p>This KE is applicable for both sexes, across developmental stages into adulthood, in numerous cells and tissues and across taxa</p>
CL:0000255eukaryotic cellHighMixedHighFoetalModerateEmbryoHighDuring development and at adulthoodHighHighHighHigh<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_1">Alapi EM, Fischer J (2006) Table of Selected Analogue Classes. In <em>Analogue-based Drug Discovery</em>, Fischer J, Ganellin CR (eds), p 515. Weinheim: Wiley-VCH Verlag GmbH & Co</a></span></span></p>
<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_3">Draskau MK, Boberg J, Taxvig C, Pedersen M, Frandsen HL, Christiansen S, Svingen T (2019) In vitro and in vivo endocrine disrupting effects of the azole fungicides triticonazole and flusilazole. <em>Environ Pollut</em> <strong>255:</strong> 113309</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_4">Foster PM, Harris MW (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>Toxicol Sci</em> <strong>85:</strong> 1024-1032</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_5">Hass U, Scholze M, Christiansen S, Dalgaard M, Vinggaard AM, Axelstad M, Metzdorff SB, Kortenkamp A (2007) Combined exposure to anti-androgens exacerbates disruption of sexual differentiation in the rat. <em>Environ Health Perspect</em> <strong>115 Suppl. 1:</strong> 122-128</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_8">Kita DH, Meyer KB, Venturelli AC, Adams R, Machado DL, Morais RN, Swan SH, Gennings C, Martino-Andrade AJ (2016) Manipulation of pre and postnatal androgen environments and anogenital distance in rats. <em>Toxicology</em> <strong>368-369:</strong> 152-161</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_9">Kjærstad MB, Taxvig C, Nellemann C, Vinggaard AM, Andersen HR (2010) Endocrine disrupting effects in vitro of conazole antifungals used as pesticides and pharmaceuticals. <em>Reprod Toxicol</em> <strong>30:</strong> 573-582</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 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"><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><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> </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 name="_ENREF_20">Vinggaard AM, Niemelä J, Wedebye EB, Jensen GE (2008) Screening of 397 chemicals and development of a quantitative structure--activity relationship model for androgen receptor antagonism. <em>Chem Res Toxicol</em> <strong>21:</strong> 813-823</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a 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>
2016-11-29T18:41:222022-06-15T06:17:59Altered, Transcription of genes by ARAltered, Transcription of genes by ARCellular<p><u>The Androgen Receptor and its function</u></p>
<p>Androgens act by binding to the Androgen receptor (AR) in androgen-responsive tissues (Davey and Grossmann 2016). Human AR mutations and mouse knockout models have established the fundamental role of 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>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). Upon activation by ligand-binding, the AR translocate 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 varies between different cells and tissues, as well as with developmental stages and is, for instance, dependent on available co-regulators (Bevan and Parker 1999; Heemers and Tindall 2007).</p>
<p>Several 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) and can for instance be found in the Androgen-Responsive Gene Database (Jiang et al. 2009).</p>
<p><em>In vitro</em></p>
<p>Decreased 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, eg transcriptomics.</p>
<p>Indirect approaches include the use of transient or stable transactivation assays including the validated OECD test guideline assay, Test No. 458: <em>Stably Transfected Human Androgen Receptor Transcriptional Activation Assay for Detection of Androgenic Agonist and Antagonist Activity of Chemicals </em>(OECD 2016). The stably transfected AR-EcoScreenTM cell line is freely available for the Japanese Collection of Research Bioresources (JCRB) Cell Bank under reference number JCRB1328. These cell-based transcriptional activation assays are typically used to detect AR agonists and antagonists. However, these types of assays are well suited to measure this KE as what they measure is exactly AR transcriptional activity. Other assays along this line 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 (van der Burg et al. 2010).</p>
<p><em>In vivo</em></p>
<p>Known downstream target gene transcription level can be measured in tissues by RT-qPCR or other gene expression analyses approaches.</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).</p>
<p>This KE is applicable for both sexes, across developmental stages into adulthood, in numerous cells and tissues and across taxa.</p>
CL:0000255eukaryotic cellHighMixedHighFoetalHighAdult, reproductively matureHighHighHigh<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>Draskau MK, Boberg J, Taxvig C, et al (2019) In vitro and in vivo endocrine disrupting effects of the azole fungicides triticonazole and flusilazole. Environ Pollut 255:113309. doi: 10.1016/j.envpol.2019.113309</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>Jiang M, Ma Y, Chen C, et al (2009) Androgen-Responsive Gene Database: Integrated Knowledge on Androgen-Responsive Genes. Mol Endocrinol 23:1927–1933. doi: 10.1210/me.2009-0103</p>
<p>Kjærstad MB, Taxvig C, Nellemann C, et al (2010) Endocrine disrupting effects in vitro of conazole antifungals used as pesticides and pharmaceuticals. Reprod Toxicol 30:573–582. doi: 10.1016/J.REPROTOX.2010.07.009</p>
<p>Laier P, Metzdorff SB, Borch J, et al (2006) Mechanisms of action underlying the antiandrogenic effects of the fungicide prochloraz. Toxicol Appl Pharmacol 213:160–71. doi: 10.1016/j.taap.2005.10.013</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</p>
<p>OECD (2016) Test No. 458: Stably Transfected Human Androgen Receptor Transcriptional Activation Assay for Detection of Androgenic Agonist and Antagonist Activity of Chemicals, OECD Guide. OECD Publishing</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>Sonneveld E, Jansen HJ, Riteco JAC, et al (2005) Development of Androgen-and Estrogen-Responsive Bioassays, Members of a Panel of Human Cell Line-Based Highly Selective Steroid-Responsive Bioassays. Toxicol Sci 83:136–148. doi: 10.1093/toxsci/kfi005</p>
<p>van der Burg B, Winter R, Man H yen, et al (2010) Optimization and prevalidation of the in vitro AR CALUX method to test androgenic and antiandrogenic activity of compounds. Reprod Toxicol 30:18–24. doi: 10.1016/j.reprotox.2010.04.012</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>
2016-11-29T18:41:232020-11-04T11:11:01Decreased fertility, Reduced number of oocytes ovulated Decreased fertility, Reduced number of oocytes ovulated Individual2016-11-29T18:41:282016-12-03T16:37:52Granulosa cell proliferation of gonadotropin-independent follicles, ReducedReduced granulosa cell proliferationCellular<p style="text-align:start"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><u><span style="font-size:12pt">Granulosa cell function</span></u></span></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><span style="font-size:12pt">Granulosa cells of the ovary play an important structural and functional role during folliculogenesis. They form the ovarian follicle architecture and transmit molecular messages to the oocyte through gap junction channels, ensuring developmental competence<span style="color:black"><span style="font-size:11pt">(Kidder and Vanderhyden, 2010)</span></span></span>. Folliculogenesis can be roughly divided into two phases: gonadotropin-independent and gonadotropin-dependent by the requirement for the gonadotropin follicle-stimulating hormone (FSH) to grow<span style="color:black"><span style="font-size:11pt">(Hsueh et al., 2015)</span></span>. During the gonadotropin-independent growth phase, growth factors secreted by the follicle, e.g. growth differentiation factor-9 (GDF9) by the oocyte and anti-Müllerian hormone (AMH) by the granulosa cells control the necessary morphological changes of granulosa cells and their proliferation<span style="color:black"><span style="font-size:11pt">(Hsueh et al., 2015)</span></span>. The growth can be histologically observed as proliferation of the granulosa cells as the flat granulosa cells of primordial follicles become cuboidal and increase in numbers<span style="color:black"><span style="font-size:11pt">(Gougeon, 2010)</span></span>. The connection between granulosa cell numbers and follicle growth during gonadotropin-independent growth is well described <span style="color:black"><span style="font-size:11pt">(Gougeon and Chainy, 1987)</span></span>.</span></span></span></p>
<p style="text-align:justify"> </p>
<p style="text-align:start"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><u><span style="font-size:12pt">Reduced granulosa cell proliferation as Key Event</span></u></span></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><span style="font-size:12pt">Genetically modified mouse models have demonstrated that granulosa cell proliferation is a prerequisite for normal follicle growth and fertility. For example, deletion of the oocyte-specific growth factor GDF9 that stimulates granulosa cells halt folliculogenesis at the primary follicle stage in mice: the granulosa cells fail to proliferate to generate secondary follicles, the oocytes degenerate, and the mice are sterile<span style="color:black"><span style="font-size:11pt">(Dong et al., 1996)</span></span></span>. Conversely, mice administered GDF9 have accelerated granulosa cell proliferation and higher numbers of primary and secondary follicles compared to non-treated ones<span style="color:black"><span style="font-size:11pt">(Vitt et al., 2000)</span></span>.</span></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><span style="font-size:12pt">AMH is a growth factor secreted by granulosa cells during the gonadotropin-independent follicle growth stage, and it inhibits the activation of primordial follicles to keep the growing and dormant follicles in balance. In mice overexpressing AMH, follicle growth to antral stages is inhibited and the numbers of all developmental stages of follicles decline faster by age than in wildtype controls<span style="color:black"><span style="font-size:11pt">(Pankhurst et al., 2018)</span></span></span>. Exposure of human ovarian tissue to AMH in culture inhibits follicle growth<span style="color:black"><span style="font-size:11pt">(Carlsson et al., 2006)</span></span>.</span></span></span></p>
<p style="text-align:start"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><em><span style="font-size:12pt">In vitro</span></em></span></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><span style="font-size:12pt">Decreased granulosa cell proliferation can be measured in cell culture. There are commercially available human granulosa cell tumor lines, for instance KGN (#RCB1154) “Granulosa cell tumor”, available from the Riken cell Bank. This cell line is representative of undifferentiated granulosa cells at early stages of follicle development making it suitable to study interactions of primordial to early antral pathways independent from hormonal control from theca cells and hypothalamic-pituitary axis <span style="color:black"><span style="font-size:11pt">(Nishi et al., 2001)</span></span></span>.</span></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><span style="font-size:12pt">Well-established assays to detect proliferation include methods to assess DNA synthesis (e.g. BrdU), cellular metabolism (e.g. MTT, XTT, ATP detection assays), and proliferation proteins (e.g. PCNA, Ki67, MCM-2)<span style="color:black"><span style="font-size:11pt">(Adan et al., 2016)</span></span></span>. The same methods can also be used in ovarian follicle or tissue culture.</span></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><em><span style="font-size:12pt">In vivo</span></em></span></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><span style="font-size:12pt">Granulosa cell proliferation manifests as increased numbers of granulosa cells within ovarian follicles<span style="color:black"><span style="font-size:11pt">(Gougeon and Chainy, 1987)</span></span></span>. Analysis of follicle growth is based on the numbers of granulosa cell layers which is also reflected in the diameter of the follicle<span style="color:black"><span style="font-size:11pt">(Gougeon and Chainy, 1987)</span></span>. Granulosa cell proliferation is inseparably connected to folliculogenesis, and therefore numbers of follicles in different developmental stages reflect the proliferation of granulosa cells. Granulosa cell proliferation can therefore be measured by counting follicles in different stages (primordial, primary, secondary) or by measuring the follicle diameters. Changes in the proliferation of granulosa cells during the early follicle growth phase would lead to altered proportions of follicles in different stages. For example, inhibition of granulosa cell proliferation can lead to reduced numbers of secondary follicles<span style="color:black"><span style="font-size:11pt">(Dong et al., 1996; Pankhurst et al., 2018)</span></span>. Therefore, studying ratios between follicles in different developmental stages can reveal changes in the proliferation of granulosa cells. Follicle counts are already suggested endpoints in the <span style="color:black">Extended One-Generation Reproductive Toxicity Study; EOGRTS (OECD 443)</span><span style="color:black"><span style="font-size:11pt">(2018)</span></span><span style="color:black">.</span></span></span></span></p>
<p style="text-align:start"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><u><span style="font-size:12pt">Overview</span></u></span></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><span style="font-size:12pt">Mechanisms controlling folliculogenesis are well conserved between mammalian species, including mice, farm animals and humans<span style="color:black"><span style="font-size:11pt">(Adhikari and Liu, 2009; McGee and Hsueh, 2000)</span></span></span>.</span></span></span></p>
<p> </p>
UBERON:0001305ovarian follicleCL:0000255eukaryotic cellHighFemaleHighDuring development and at adulthoodHighHighHighHighHighHigh<p style="text-align:start"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000">Adan, A., Kiraz, Y., and Baran, Y. (2016). Cell Proliferation and Cytotoxicity Assays. Current Pharmaceutical Biotechnology <em>17</em>, 1213–1221. https://doi.org/10.2174/1389201017666160808160513.</span></span></span></p>
<p style="text-align:start"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000">Adhikari, D., and Liu, K. (2009). Molecular mechanisms underlying the activation of mammalian primordial follicles. Endocrine Reviews <em>30</em>, 438–464. https://doi.org/10.1210/er.2008-0048.</span></span></span></p>
<p style="text-align:start"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000">Carlsson, I.B., Scott, J.E., Visser, J.A., Ritvos, O., Themmen, A.P.N., and Hovatta, O. (2006). Anti-Müllerian hormone inhibits initiation of growth of human primordial ovarian follicles in vitro. Human Reproduction <em>21</em>, 2223–2227. https://doi.org/10.1093/humrep/del165.</span></span></span></p>
<p style="text-align:start"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000">Dong, J., Albertini, D.F., Nishimori, K., Kumar, T.R., Lu, N., and Matzuk, M.M. (1996). Growth differentiation factor-9 is required during early ovarian folliculogenesis. Nature <em>383</em>, 531–535. https://doi.org/10.1038/383531a0.</span></span></span></p>
<p style="text-align:start"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000">Gougeon, A. (2010). Croissance folliculaire dans l’ovaire humain: de l’entrée en croissance du follicule primordial jusqu’à la maturation préovulatoire. Annales d’Endocrinologie <em>71</em>, 132–143. https://doi.org/10.1016/j.ando.2010.02.021.</span></span></span></p>
<p style="text-align:start"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000">Gougeon, A., and Chainy, G.B.N. (1987). Morphometric studies of small follicles in ovaries of women at different ages. Journal of Reproduction and Fertility <em>81</em>, 433–442. https://doi.org/10.1530/jrf.0.0810433.</span></span></span></p>
<p style="text-align:start"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000">Hsueh, A.J.W., Kawamura, K., Cheng, Y., and Fauser, B.C.J.M. (2015). Intraovarian control of early folliculogenesis. Endocrine Reviews <em>36</em>, 1–24. https://doi.org/10.1210/er.2014-1020.</span></span></span></p>
<p style="text-align:start"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000">Kidder, G.M., and Vanderhyden, B.C. (2010). Bidirectional communication between oocytes and follicle cells: Ensuring oocyte developmental competence. Canadian Journal of Physiology and Pharmacology <em>88</em>, 399–413. https://doi.org/10.1139/Y10-009.</span></span></span></p>
<p style="text-align:start"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000">McGee, E.A., and Hsueh, A.J.W. (2000). Initial and Cyclic Recruitment of Ovarian Follicles*. Endocrine Reviews <em>21</em>, 200–214. https://doi.org/10.1210/edrv.21.2.0394.</span></span></span></p>
<p style="text-align:start"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000">Nishi, Y., Yanase, T., Mu, Y.-M., Oba, K., Ichino, I., Saito, M., Nomura, M., Mukasa, C., Okabe, T., Goto, K., et al. (2001). Establishment and Characterization of a Steroidogenic Human Granulosa-Like Tumor Cell Line, KGN, That Expresses Functional Follicle-Stimulating Hormone Receptor. Endocrinology <em>142</em>, 437–445. https://doi.org/10.1210/endo.142.1.7862.</span></span></span></p>
<p style="text-align:start"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000">Pankhurst, M.W., Kelley, R.L., Sanders, R.L., Woodcock, S.R., Oorschot, D.E., and Batchelor, N.J. (2018). Anti-Müllerian hormone overexpression restricts preantral ovarian follicle survival. Journal of Endocrinology <em>237</em>, 153–163. https://doi.org/10.1530/JOE-18-0005.</span></span></span></p>
<p style="text-align:start"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000">Vitt, U.A., McGee, E.A., Hayashi, M., and Hsueh, A.J.W. (2000). In vivo treatment with GDF-9 stimulates primordial and primary follicle progression and theca cell marker CYP17 in ovaries of immature rats. Endocrinology <em>141</em>, 3814–3820. https://doi.org/10.1210/endo.141.10.7732.</span></span></span></p>
<p style="text-align:start"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000">(2018). Test No. 443: Extended One-Generation Reproductive Toxicity Study (OECD).</span></span></span></p>
2020-10-13T10:19:022022-04-20T10:42:33343d418c-1b59-4f82-b409-aee2df1036a843af122c-a285-469d-968a-641cdae46c7f<p><span style="font-family:calibri,sans-serif; font-size:11.0pt">The AR is a nuclear hormone receptor that functions primarily as a transcription factor. After binding of a ligand (androgens: DHT, testosterone) to the AR, the androgen-AR complex translocate to the nucleus and bind to specific hormone response elements on the DNA duplex to regulate gene transcription. The AR is expressed in various cells and tissues throughout the body in a spatiotemporal manner, with levels and activation changed in response to androgen signaling </span><!--[if supportFields]><span
lang=EN-US style='font-size:11.0pt;line-height:107%;font-family:"Calibri",sans-serif;
mso-ascii-theme-font:minor-latin;mso-fareast-font-family:Calibri;mso-fareast-theme-font:
minor-latin;mso-hansi-theme-font:minor-latin;mso-bidi-font-family:"Times New Roman";
mso-bidi-theme-font:minor-bidi;mso-ansi-language:EN-US;mso-fareast-language:
EN-US;mso-bidi-language:AR-SA'><span style='mso-element:field-begin'></span><span
style='mso-spacerun:yes'> </span>ADDIN EN.CITE <span style='mso-element:field-begin'></span><span
style='mso-spacerun:yes'> </span>ADDIN EN.CITE.DATA <![if gte mso 9]><xml>
<w:data>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</w:data>
</xml><![endif]><span style='mso-element:field-end'></span><span
style='mso-element:field-separator'></span></span><![endif]--><span style="font-family:calibri,sans-serif; font-size:11.0pt">(<a href="#_ENREF_1" title="Chang, 1995 #271"><span style="color:windowtext">Chang et al, 1995</span></a>; <a href="#_ENREF_2" title="Davey, 2016 #250"><span style="color:windowtext">Davey & Grossmann, 2016</span></a>; <a href="#_ENREF_20" title="Roy, 1999 #270"><span style="color:windowtext">Roy et al, 1999</span></a>)<!--[if gte mso 9]><xml>
<w:data>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</w:data>
</xml><![endif]--></span><!--[if supportFields]><span lang=EN-US
style='font-size:11.0pt;line-height:107%;font-family:"Calibri",sans-serif;
mso-ascii-theme-font:minor-latin;mso-fareast-font-family:Calibri;mso-fareast-theme-font:
minor-latin;mso-hansi-theme-font:minor-latin;mso-bidi-font-family:"Times New Roman";
mso-bidi-theme-font:minor-bidi;mso-ansi-language:EN-US;mso-fareast-language:
EN-US;mso-bidi-language:AR-SA'><span style='mso-element:field-end'></span></span><![endif]--><span style="font-family:calibri,sans-serif; font-size:11.0pt">.</span></p>
<p><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">The fact that AR exert its primary function as a nuclear transcription factor is well-established and a generally accepted fact. Binding of androgens (ligands) to the AR induces receptor activation and transcriptional regulation of target genes </span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">(</span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"><a href="#_ENREF_1" title="Chang, 1995 #271">Chang et al, 1995</a></span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">; </span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"><a href="#_ENREF_2" title="Davey, 2016 #250">Davey & Grossmann, 2016</a></span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">; </span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"><a href="#_ENREF_6" title="Heemers, 2007 #255">Heemers & Tindall, 2007</a></span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">)</span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">. After binding to Androgen Response Elements (AREs), the AR can recruit a variety of co-regulators (activators or repressors) that will influence the transcriptional regulation of target genes and thereby achieve spatiotemporally regulated gene expression </span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">(</span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"><a href="#_ENREF_6" title="Heemers, 2007 #255">Heemers & Tindall, 2007</a></span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">)</span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">. Consequently, inhibition/competition of ligand binding to the AR leading to reduced activity will have an effect on downstream transcriptional function.</span></span></p>
<p><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">The fact that AR exert its primary function as a nuclear transcription factor is well-established and a generally accepted fact. Binding of androgens (ligands) to the AR induces receptor activation and transcriptional regulation of target genes </span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">(</span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"><a href="#_ENREF_1" title="Chang, 1995 #271">Chang et al, 1995</a></span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">; </span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"><a href="#_ENREF_2" title="Davey, 2016 #250">Davey & Grossmann, 2016</a></span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">; </span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"><a href="#_ENREF_6" title="Heemers, 2007 #255">Heemers & Tindall, 2007</a></span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">)</span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">. After binding to Androgen Response Elements (AREs), the AR can recruit a variety of co-regulators (activators or repressors) that will influence the transcriptional regulation of target genes and thereby achieve spatiotemporally regulated gene expression </span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">(</span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"><a href="#_ENREF_6" title="Heemers, 2007 #255">Heemers & Tindall, 2007</a></span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">)</span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">. Consequently, inhibition/competition of ligand binding to the AR leading to reduced activity will have an effect on downstream transcriptional function.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">The activation of AR by ligand binding is crucial for transcriptional activation of target genes. Thus, antagonism of AR (upstream KE) will have a direct effects on nuclear translocation and subsequent transcriptional activation of target genes (downstream KE). There is ample evidence showing that AR regulate gene expression in a large number of cells and tissues across animal species, as well as altered gene expression following ectopic activation or antagonisms of AR (<a href="#_ENREF_5" title="Grosse, 2012 #74">Grosse et al, 2012</a>; <a href="#_ENREF_10" title="Lamb, 2014 #75">Lamb et al, 2014</a>; <a href="#_ENREF_11" title="Lamont, 2010 #77">Lamont & Tindall, 2010</a>; <a href="#_ENREF_13" title="Matsumoto, 2013 #76">Matsumoto et al, 2013</a>; <a href="#_ENREF_14" title="Mikkonen, 2010 #71">Mikkonen et al, 2010</a>; <a href="#_ENREF_18" title="Olsen, 2016 #72">Olsen et al, 2016</a>). Below are examples of evidence for AR antagonism leading to altered gene transactivation (assay) or expression of gene targets in cells and tissues. The list is not exhaustive.</span></span></p>
<ul>
<li><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">There are several drugs that bind the AR ligand binding domain in competition to androgens (testosterone and DHT), thereby inhibiting AR nuclear translocation and transcriptional activation, for instance flutamide (<a href="#_ENREF_7" title="Irwin, 1973 #275">Irwin & Prout Jr, 1973</a>), biculamide (<a href="#_ENREF_3" title="Furr, 1988 #276">Furr, 1988</a>), enzalutamide (<a href="#_ENREF_25" title="Tran, 2009 #274">Tran et al, 2009</a>) and darolutamide (<a href="#_ENREF_15" title="Moilanen, 2015 #66">Moilanen et al, 2015</a>). </span></span></li>
<li><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Exploiting well-characterized AR response elements (AREs), several in vitro AR reporter (transactivation) assays have been developed, and reported on extensively in the scientific literature. These study show direct antagonistic effects of a large number of compounds on the AR, with several examples listed under KE 26. </span></span></li>
<li><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Using ChIP and DNA/RNA sequencing technologies, a complex network of genes directly regulated by AR has been mapped in for instance prostate cancer cells (<a href="#_ENREF_23" title="Takayam, 2013 #68">Takayam & Inoue, 2013</a>).</span></span></li>
<li><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Plumagin, a naphthoquinone, can inhibit DHT-mediated AR-regulated gene expression in prostate cancer cells, shown by RNA-seq analysis (<a href="#_ENREF_19" title="Rondeau, 2018 #67">Rondeau et al, 2018</a>).</span></span></li>
<li><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Short-term exposure to the AR antagonists flutamide and vinclozolin significantly alters the gonadal transcriptome in mature zebrafish (<a href="#_ENREF_12" title="Martinović-Weigelt, 2011 #69">Martinović-Weigelt et al, 2011</a>)</span></span></li>
<li><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">In utero exposure to rats to the AR antagonist finasteride alters the transcriptome in male perineal tissues (<a href="#_ENREF_22" title="Schwartz, 2019 #51">Schwartz et al, 2019</a>).</span></span></li>
<li><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Genome-wide analysis of AR target genes in mesenchymal cells during human prostate development has shown variable expression of AR target genes in androgen insensitivity syndrome patients (<a href="#_ENREF_16" title="Nash, 2019 #70">Nash et al, 2019</a>). </span></span></li>
<li><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">Human LNCaP prostate cancer cells stimulated for 24h with the AR agonist R1881 display up- and down-regulation of around 300 genes in both directions </span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">(</span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"><a href="#_ENREF_17" title="Ngan, 2009 #277">Ngan et al, 2009</a></span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">)</span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">.</span></span></li>
</ul>
<p><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">The AR gene contains CAG repeats (encoding for the amino acid glutamine), which vary between individuals and will affect transcriptional function. Broadly speaking, fewer CAG repeats tend to render the AR more sensitive to androgen activation whereas more CAG repeats tend to render the AR less sensitive, albeit the functional relevance at the tissue/organ level remains less clear </span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">(</span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"><a href="#_ENREF_24" title="Tirabassi, 2015 #273">Tirabassi et al, 2015</a></span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">; </span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"><a href="#_ENREF_28" title="Zitzmann, 2009 #272">Zitzmann, 2009</a></span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">)</span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">. It is plausible, however, that it may lead to variable sensitivity to AR antagonism. </span></span></p>
<p><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">The quantitative relationship between AR antagonism and transcriptional activity can be measured in vitro through several available reporter assay such as AR-EcoScreen </span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">(</span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"><a href="#_ENREF_21" title="Satoh, 2004 #280">Satoh et al, 2004</a></span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">)</span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"> and AR-CALUX </span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">(</span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"><a href="#_ENREF_26" title="van der Burg, 2010 #261">van der Burg et al, 2010</a></span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">)</span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">, different validated reporters assays </span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">(</span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"><a href="#_ENREF_9" title="Körner, 2004 #282">Körner et al, 2004</a></span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">)</span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"> plus several more. Dose-response curve assays are performed, allowing for the calculation of potential to antagonize AR (e.g. EC<sub>50 </sub>scores, log10 Ki values). There is a large body of studies having reported on AR antagonism by numerous chemicals </span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">(</span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"><a href="#_ENREF_4" title="Gray, 2020 #278">Gray et al, 2020</a></span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">; </span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"><a href="#_ENREF_8" title="Kleinstreuer, 2017 #281">Kleinstreuer et al, 2017</a></span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">; </span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"><a href="#_ENREF_27" title="Vinggaard, 2008 #263">Vinggaard et al, 2008</a></span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">)</span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">. </span></span></p>
HighMaleHighFemaleHighAll life stagesHighHighHighHigh<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_1">Chang AY, Saltzman A, Yeh S, Young W, Keller E, Lee HJ, Wang HJ, Mizokami A (1995) Androgen receptor: an overview. <em>Critical Reviews in Eukaryotic Gene Expression</em> <strong>5:</strong> 97-125</a></span></span></p>
<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>Clinical Biochemist Reviews</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_3">Furr BJ (1988) ICI 176,334: a novel non-steroidal, peripherally-selective antiandrogen. <em>Prog Clin Biol Res</em> <strong>260:</strong> 13-26</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_4">Gray LE, Furr JR, Lambright CS, Evans N, Hartig PC, Cardon MC, Wilson VS, Hotchkiss AK, Conley JM (2020) Quantification of the Uncertainties in Extrapolating From In Vitro Androgen Receptor Antagonism to In Vivo Hershberger Assay Endpoints and Adverse Reproductive Development in Male Rats. <em>Toxicol Sci</em> <strong>176:</strong> 297-311</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_5">Grosse A, Bartsch S, Baniahmad A (2012) Androgen receptor-mediated gene repression. <em>Mol Cell Endocrinol</em> <strong>352:</strong> 46-56</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">Irwin RJ, Prout Jr GR (1973) A new antiprostatic agent for treatment of prostatic carcinoma. <em>Surg Forum</em> <strong>24:</strong> 536-537</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_8">Kleinstreuer NC, Ceger P, Watt ED, Martin M, Houck K, Browne P, Thomas RS, Casey WM, Dix DJ, Allen D, Sakamuru S, Xia M, Huang R, Judson R (2017) Development and Validation of a Computational Model for Androgen Receptor Activity. <em>Chem Res Toxicol</em> <strong>30:</strong> 946-964</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_9">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_10">Lamb AD, Massie CE, Neal DE (2014) The transcriptional programme of the androgen receptor (AR) in prostate cancer. <em>BJU Int</em> <strong>113:</strong> 358-366</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_11">Lamont KR, Tindall DJ (2010) Androgen regulation of gene expression. <em>Adv Cancer Res</em> <strong>107:</strong> 137-162</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_12">Martinović-Weigelt D, Wang RL, Villeneuve DL, Bencic DC, Lazorchak J, Ankley GT (2011) Gene expression profiling of the androgen receptor antagonists flutamide and vinclozolin in zebrafish (Danio rerio) gonads. <em>Aquat Toxicol</em> <strong>101:</strong> 447-458</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_13">Matsumoto T, Sakari M, Okada M, Yokoyama A, Takahashi S, Kouzmenko A, Kato S (2013) The androgen receptor in health and disease. <em>Annu Rev Physiol</em> <strong>75:</strong> 201-224</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_14">Mikkonen L, Pihlajamaa P, Sahu B, Zhang FP, Jänne OA (2010) Androgen receptor and androgen-dependent gene expression in lung. <em>Mol Cell Endocrinol</em> <strong>317:</strong> 14-24</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_15">Moilanen AM, Riikonen R, Oksala R, Ravanti L, Aho E, Wohlfahrt G, Nykänen PS, Törmäkangas OP, Palvimo JJ, Kallio PJ (2015) Discovery of ODM-201, a new-generation androgen receptor inhibitor targeting resistance mechanisms to androgen signaling-directed prostate cancer therapies. <em>Sci Rep</em> <strong>5:</strong> 12007</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_16">Nash C, Boufaied N, Badescu D, Wang YC, Paliouras M, Trifiro M, Ragoussis I, Thomson AA (2019) Genome-wide analysis of androgen receptor binding and transcriptomic analysis in mesenchymal subsets during prostate development. <em>Dis Model Mech</em> <strong>12:</strong> dmm039297</a></span></span></p>
<p> </p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_17">Ngan S, Stronach EA, Photiou A, Waxman J, Ali S, Buluwela L (2009) Microarray coupled to quantitative RT–PCR analysis of androgen-regulated genes in human LNCaP prostate cancer cells. <em>Oncogene</em> <strong>28:</strong> 2051-2063</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_18">Olsen JR, Azeem W, Hellem MR, Marvyin K, Hua Y, Qu Y, Li L, Lin B, Ke XS, Øyan AM, Kalland KH (2016) Context dependent regulatory patterns of the androgen receptor and androgen receptor target genes. <em>BMC Cancer</em> <strong>16:</strong> 377</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_19">Rondeau G, Abedinpour P, Chrastina A, Pelayo J, Borgstrom P, Welsh J (2018) Differential gene expression induced by anti-cancer agent plumbagin is mediated by androgen receptor in prostate cancer cells. <em>Sci Rep</em> <strong>8:</strong> 2694</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_20">Roy AK, Lavrovsky Y, Song CS, Chen S, Jung MH, Velu NK, Bi BY, Chatterjee B (1999) Regulation of androgen action. <em>Vitam Horm</em> <strong>55:</strong> 309-352</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_21">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><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_22">Schwartz CL, Vinggaard AM, Christiansen S, Darde TA, Chalmel F, Svingen T (2019) Distinct transcriptional profiles of the female, male and finasteride-induced feminized male anogenital region in rat fetuses. <em>Toxicol Sci</em> <strong>169:</strong> 303-311</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_23">Takayam K, Inoue S (2013) Transcriptional network of androgen receptor in prostate cancer progression. <em>Int J Urol</em> <strong>20:</strong> 756-768</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_24">Tirabassi G, Cignarelli A, Perrini S, Delli Muti N, Furlani G, Gallo M, Pallotti F, Paoli D, Giorgino F, Lombardo F, Gandini L, Lenzi A, Balercia G (2015) Influence of CAG Repeat Polymorphism on the Targets of Testosterone Action. <em>Int J Endocrinol</em> <strong>2015:</strong> 298107</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_25">Tran C, Ouk S, Clegg NJ, Chen Y, Watson PA, Arora V, Wongvipat J, Smith-Jones PM, Yoo D, Kwon A, Wasielewska T, Welsbie D, Chen CD, Higano CS, Beer TM, Hung DT, Scher HI, Jung ME, Sawyers CL (2009) Development of a second-generation antiandrogen for treatment of advanced prostate cancer. <em>Science</em> <strong>324:</strong> 787-790</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_26">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 name="_ENREF_27">Vinggaard AM, Niemelä J, Wedebye EB, Jensen GE (2008) Screening of 397 chemicals and development of a quantitative structure--activity relationship model for androgen receptor antagonism. <em>Chem Res Toxicol</em> <strong>21:</strong> 813-823</a></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><a name="_ENREF_28">Zitzmann M (2009) The role of the CAG repeat androgen receptor polymorphism in andrology. <em>Front Horm Res</em> <strong>37:</strong> 52-61</a></span></span></p>
2020-11-02T07:51:382021-02-02T05:02:4243af122c-a285-469d-968a-641cdae46c7f2a53b981-71cd-49da-aea8-3a52bc527a1a<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><span style="font-size:12pt">Decreased transcription of genes that are downstream of AR activation leads to reduced granulosa cell proliferation of the early-stage gonadotropin-independent ovarian follicles. Therefore, the follicle growth to the antral stage is decreased.</span></span></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><span style="font-size:12pt">AR is a ligand-activated nuclear transcription factor expressed in the ovaries across mammalian species, including humans<span style="color:black"><span style="font-size:11pt">(Gervásio et al., 2014)</span></span></span>. During the gonadotropin independent follicular stage, AR activation is hypothesized to promote follicle growth, whereas in later stages it has been shown to inhibit growth and induce apoptosis<span style="color:black"><span style="font-size:11pt">(Franks and Hardy, 2018; Harlow et al., 1988)</span></span>. In humans, both mRNA and protein of AR are present in the oocyte, stroma cells, theca cells, but most prominently in granulosa cells of small antral follicles<span style="color:black"><span style="font-size:11pt">(Gervásio et al., 2014; Jeppesen et al., 2012)</span></span>. </span></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><span style="font-size:12pt">In the mouse ovary, AR mRNA and protein are present in the oocyte, theca, and granulosa cells<span style="color:black"><span style="font-size:11pt">(Gill et al., 2004; Hirai et al., 1994; Szoltys and Slomczynska, 2000; Tetsuka and Hillier, 1996; Tetsuka et al., 1995)</span></span></span>. In the cow and sheep ovary, AR mRNA is present in granulosa and theca cells, and most prominently in granulosa of antral and early antral follicles<span style="color:black"><span style="font-size:11pt">(Hampton et al., 2004; Juengel et al., 2006)</span></span>. In the pig ovary, AR mRNA is mainly expressed in the granulosa cells until the antral stage<span style="color:black"><span style="font-size:11pt">(Cárdenas and Pope, 2002; Slomczynska et al., 2001)</span></span>. In the monkey ovary, AR mRNA and protein are present in theca, but mainly granulosa cells of antral and early antral follicles<span style="color:black"><span style="font-size:11pt">(Hillier et al., 1997; Weil et al., 1998)</span></span>.</span></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><span style="font-size:12pt">In human follicles, the expression of the <em>AR</em> transcript is observed after the primordial stage and is most pronounced during the small antral stage<span style="color:black"><span style="font-size:11pt">(Rice et al., 2007)</span></span></span>. Throughout early folliculogenesis, AR expression controls transcription of genes involved in promoting growth and differentiation of granulosa cells and formation of antrum<span style="color:black"><span style="font-size:11pt">(Gervásio et al., 2014)</span></span>. Genes under the control of AR that are involved in these processes include Kit ligand (<em>KITL</em>), Bone morphogenetic protein 15 (<em>BMP15</em>), and Hepatocyte growth factor (<em>HGF</em>)<span style="color:black"><span style="font-size:11pt">(Astapova et al., 2019; Prizant et al., 2014)</span></span>.</span></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><span style="font-size:12pt">In the monkey ovary, high levels of <em>AR</em> mRNA correlates with high levels of granulosa cell proliferation<span style="color:black"><span style="font-size:11pt">(Vendola et al., 1998; Weil et al., 1998)</span></span></span> Increased AR activation is associated with increased follicle growth and increased granulosa cell proliferation in small antral rat follicles, supporting the important role for AR during this developmental stage<span style="color:black"><span style="font-size:11pt">(Lim et al., 2017a)</span></span>.</span></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><span style="font-size:12pt">AR may mediate early follicle growth through FSHR, supported by studies correlating mRNA levels of AR and FSHR in granulosa cells of small antral follicles<span style="color:black"><span style="font-size:11pt">(Nielsen et al., 2011,</span></span></span><sup>,</sup><span style="color:black"><span style="font-size:11pt">Weil et al., 1999)</span></span>. In mice, FSH-mediated <em>in vitro</em> follicle growth is increased by androgens, suggesting that androgens through AR may act synergistically with FSHR, which in turn increases follicle growth to antral follicles<span style="color:black"><span style="font-size:11pt">(Sen et al., 2014)</span></span>.</span></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><span style="font-size:12pt">It is also hypothesized that FSHR activation through AR leads to increased AMH expression in granulosa cells of primary to small antral follicles<span style="color:black"><span style="font-size:11pt">(Lin et al., 2021)</span></span></span>. In turn, elevated levels of AMH lead to inhibition of FSH-induced aromatase activity, resulting in higher androgen levels that inhibit further follicular growth.</span></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><span style="font-size:12pt">AR activation has been associated with Insulin-like Growth Factor 1 (IGF1) and Insulin-like Growth Factor 1 Receptor (IGFR1) and other key factors of the IGF signaling pathway, which is essential for granulosa growth and differentiation<span style="color:black"><span style="font-size:11pt">(Baumgarten et al., 2014)</span></span></span><sup>,</sup><span style="color:black"><span style="font-size:11pt">(Vendola et al., 1999)</span></span>. In human granulosa cells of primordial and primary follicles, AR and IGF-related factors are highly enriched at the transcriptional level<span style="color:black"><span style="font-size:11pt">(Steffensen et al., 2018)</span></span>.</span></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><span style="font-size:12pt">AR activation affects the level of connexins, proteins that form gap junctions between granulosa cells and the oocyte and hence regulate intracellular communication; a prerequisite for folliculogenesis<span style="color:black"><span style="font-size:11pt">(Kamal et al., 2020)</span></span></span>.</span></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><span style="font-size:12pt">In humans, the importance of AR in follicular growth becomes evident with the beneficial effects of androgens in assisted reproductive technology outcomes<span style="color:black"><span style="font-size:11pt">(Bosdou et al., 2012; Casson et al., 2000; Fábregues et al., 2009; Kim et al., 2011, 2014; Nagels et al., 2015; Noventa et al., 2019; Petya Andreeva, Ivelina Oprova, Luboslava Valkova, Petya Chaveeva, Ivanka Dimova, 2020)</span></span></span>. Although the mechanism remains elusive, it has been suggested that androgen priming of women seeking fertility treatment promotes follicle growth resulting in an increase in the FSH-sensitive follicle pool<span style="color:black"><span style="font-size:11pt">(Hu et al., 2017)</span></span>. Gene expression studies in human small antral follicles reveal significant association of AR and FSHR levels, suggesting that the increase in follicle growth could be mediated through regulating AR transcription in granulosa cells<span style="color:black"><span style="font-size:11pt">(Hu et al., 2017; Nielsen et al., 2011)</span></span>. Epidemiological studies have shown that upon androgen pretreatment, increase in the number of antral follicles and mean follicular diameter were observed<span style="color:black"><span style="font-size:11pt">(Balasch et al., 2006; Kim et al., 2011)</span></span>. This increase supports the hypothesis that androgen receptor signaling is important for early follicle growth. Studies observing no effects upon androgen pre-treatment claim that dose and duration of the selected androgen might lead to contradicting results<span style="color:black"><span style="font-size:11pt">(Yeung et al., 2014)</span></span>.</span></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><span style="font-size:12pt">Hypoandrogenism provides further evidence for an important role for androgen actions in human follicle development. Lower levels of DHEA or testosterone have been associated with women that have diminished ovarian reserve or premature ovarian aging<span style="color:black"><span style="font-size:11pt">(Gleicher et al., 2013)</span></span></span>. In the case of untreated primary adrenal insufficiency, the androgen deficient patient exhibit significantly reduced fertility<span style="color:black"><span style="font-size:11pt">(Erichsen et al., 2010)</span></span>.</span></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><span style="font-size:12pt">Conclusions on the androgen significance can also be drawn from clinical evidence where women are exposed to an androgen excess. Hyperandrogenism in the case of congenital adrenal hyperplasia and exogenous androgen treatments in trans males lead to polycystic ovaries<span style="color:black"><span style="font-size:11pt">(Walters and Handelsman, 2018)</span></span></span>. This indicated that the androgens stimulate early follicle growth and inhibit further maturation<span style="color:black"><span style="font-size:11pt">(Walters and Handelsman, 2018)</span></span>. In polycystic ovarian syndrome, a syndrome characterized by accumulation of small antral follicles in the ovarian cortex, a plausible cause for this morphology is hyperandrogenaemia<span style="color:black"><span style="font-size:11pt">(Balen et al., 2003; Lebbe and Woodruff, 2013)</span></span>.</span></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:12.0pt">Androgen Receptor Knock Out (ARKO) mouse model</span></strong></span></span></p>
<table align="center" cellspacing="0" class="MsoTableGrid" style="border-collapse:collapse; border:none; width:110.0%">
<tbody>
<tr>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:2px solid black; vertical-align:top; width:120px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:9.0pt">Granulosa-specific ARKO model</span></strong></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:2px solid black; vertical-align:top; width:324px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:9.0pt">Relevant observations</span></strong></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:2px solid black; vertical-align:top; width:132px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:9.0pt">Reference</span></strong></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:120px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">GCARKO<sup>Ex2</sup></span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:324px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Premature ovarian failure, subfertility, longer estrous cycles, slower <em>in vitro </em>follicle growth compared to wild type</span></span></span></p>
<p style="text-align:justify"> </p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:132px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">(Sen & Hammes, 2010)</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:120px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">GCARKO<sup>Ex3</sup></span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:324px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Subfertility, longer estrous cycles, decreased number of preantral and antral follicles compared to wild type, trend of lower ovarian expression of kitl, ifgr1 and fshr compared to wild type (not statistically significant)</span></span></span></p>
<p style="text-align:justify"> </p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:132px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">(Kirsty A. Walters et al., 2012)</span></span></span></p>
</td>
</tr>
</tbody>
</table>
<p style="text-align:justify"> </p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><em><span style="font-size:12.0pt">In vitro</span></em></strong><strong><span style="font-size:12.0pt">/<em>Ex vivo</em></span></strong></span></span></p>
<table align="center" cellspacing="0" class="MsoTableGrid" style="border-collapse:collapse; border:none; width:110.0%">
<tbody>
<tr>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:2px solid black; vertical-align:top; width:81px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:9.0pt">Study type</span></strong></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:2px solid black; vertical-align:top; width:51px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:9.0pt">Species</span></strong></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:2px solid black; vertical-align:top; width:105px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:9.0pt">Compound</span></strong></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:2px solid black; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:9.0pt">Effect Dose</span></strong></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:2px solid black; vertical-align:top; width:59px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:9.0pt">Duration</span></strong></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:2px solid black; vertical-align:top; width:72px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:9.0pt">Method</span></strong></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:2px solid black; vertical-align:top; width:103px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:9.0pt">Result</span></strong></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:2px solid black; vertical-align:top; width:92px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:9.0pt">Reference</span></strong></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:81px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Isolated secondary follicles in culture</span></span></span></p>
<p style="text-align:center"> </p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:51px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Mouse</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:105px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Hydroxyflutamide,</span></span></span></p>
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Bicalutamide</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">50</span><span style="font-size:9.0pt">μΜ</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:59px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">8d, 12d</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:72px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Follicle diameter measured </span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:103px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Reduced follicular growth </span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:92px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">(Lenie & Smitz, 2009)</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:81px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Isolated late secondary follicles in culture</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:51px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Mouse</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:105px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Bicalutamide</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">1</span><span style="font-size:9.0pt">0μΜ, 32μΜ</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:59px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">6d</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:72px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Follicle diameter measured</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:103px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Reduced follicular growth (dose-depended)</span></span></span></p>
<p style="text-align:justify"> </p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:92px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">(Murray, Gosden, Allison, & Spears, 1998)</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:81px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Isolated secondary follicles in culture</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:51px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Mouse</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:105px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Hydroxyflutamide</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">1</span><span style="font-size:9.0pt">μΜ, 10μΜ</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:59px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">4d</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:72px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Follicle diameter measured</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:103px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Reduced androgen-induced follicular growth</span></span></span></p>
<p style="text-align:justify"> </p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:92px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">(Wang et al., 2001)</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:81px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Isolated secondary follicles in culture</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:51px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Mouse</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:105px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Flutamide</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">20</span><span style="font-size:9.0pt">μΜ</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:59px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">2d, 3d</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:72px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Follicle diameter and area measured</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:103px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Reduced androgen –induced follicular growth </span></span></span></p>
<p style="text-align:justify"> </p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:92px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">(Laird et al., 2017)</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:81px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Isolated secondary follicles in culture</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:51px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Mouse</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:105px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Enzalutamide</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">1</span><span style="font-size:9.0pt">μΜ</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:59px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">2d, 4d, 6d</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:72px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Follicle diameter measured and number of antral follicles counted</span></span></span></p>
<p style="text-align:justify"> </p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:103px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Reduced follicular growth and antrum formation</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:92px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">(Lebbe et al., 2017)</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:81px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Fetal ovaries in culture</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:51px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Mouse</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:105px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Vinclozolin</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">10</span><span style="font-size:9.0pt">μΜ</span><span style="font-size:9.0pt">, 50</span><span style="font-size:9.0pt">μΜ</span><span style="font-size:9.0pt">, 100</span><span style="font-size:9.0pt">μΜ</span><span style="font-size:9.0pt">, 200</span><span style="font-size:9.0pt">μΜ</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:59px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">7d of 17d </span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:72px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Follicle diameter measured</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:103px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Reduced follicular growth</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:92px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">(González-Sanz, Barreñada, Rial, Brieño-Enriquez, & del Mazo, 2020)</span></span></span></p>
<p style="text-align:justify"> </p>
</td>
</tr>
<tr>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:81px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Isolated late secondary follicles in culture</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:51px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Rat</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:105px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Flutamide</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">10</span><span style="font-size:9.0pt">μΜ</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:59px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">2d, 4d</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:72px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Follicle diameter measured and expressed as follicular volume</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:103px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Reduced GDF9 and INSL3 induced follicular growth</span></span></span></p>
<p style="text-align:justify"> </p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:92px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">(Xue, Kim, Liu, & Tsang, 2014)</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:81px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Isolated late secondary follicles in culture</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:51px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Rat</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:105px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Flutamide</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">10</span><span style="font-size:9.0pt">μΜ</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:59px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">2d, 4d</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:72px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Follicle diameter measured and expressed as follicular volume</span></span></span></p>
<p style="text-align:justify"> </p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:103px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Reduced NR4A1-induced follicular growth</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:92px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">(Xue, Liu, Murphy, & Tsang, 2012)</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:81px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Isolated late secondary follicles in culture</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:51px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Rat</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:105px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Flutamide</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">10</span><span style="font-size:9.0pt">μΜ</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:59px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">4d</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:72px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Follicle diameter measured and expressed as follicular volume</span></span></span></p>
<p style="text-align:justify"> </p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:103px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Reduced GDF9-induced follicle growth</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:92px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">(Orisaka, Jiang, Orisaka, Kotsuji, & Tsang, 2009)</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:81px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Ovarian cortex pieces in culture</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:51px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Cow</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:105px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Flutamide</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">0.1</span><span style="font-size:9.0pt">μΜ</span><span style="font-size:9.0pt">, 1</span><span style="font-size:9.0pt">μΜ</span><span style="font-size:9.0pt">, 10</span><span style="font-size:9.0pt">μΜ</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:59px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">10d</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:72px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Follicle counting and classification (histology)</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:103px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Reduced number of secondary follicles compared to testosterone group (dose-dependent)</span></span></span></p>
<p style="text-align:justify"> </p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:92px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">(Yang & Fortune, 2006)</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:81px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Isolated granulosa cells from antral follicles</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:51px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Pig</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:105px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Hydroxyflutamide</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">5</span><span style="font-size:9.0pt">μΜ</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:59px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">1d</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:72px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Incorporation of [<sup>3</sup>H]-thymidine </span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:103px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Reduced granulosa cell proliferation</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:92px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">(T. E. Hickey, Marrocco, Gilchrist, Norman, & Armstrong, 2004)</span></span></span></p>
<p style="text-align:justify"> </p>
</td>
</tr>
<tr>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:81px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Isolated mural granulosa cells from small antral follicles</span></span></span></p>
<p style="text-align:center"> </p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:51px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Pig</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:105px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Hydroxyflutamide</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">0.1</span><span style="font-size:9.0pt">μΜ</span><span style="font-size:9.0pt">, 1</span><span style="font-size:9.0pt">μΜ</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:59px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">1h of 24h</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:72px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Incorporation of [<sup>3</sup>H]-thymidine </span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:103px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Reduced granulosa cell proliferation</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:92px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">(T. E. Hickey et al., 2005)</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:81px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Ovarian cortex pieces in culture</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:51px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Pig</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:105px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Cyproterone acetate</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">0.001 </span><span style="font-size:9.0pt">μΜ</span><span style="font-size:9.0pt">,</span></span></span></p>
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">0.0001 </span><span style="font-size:9.0pt">μΜ</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:59px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">7d</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:72px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Follicle counting and classification (histology)</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:103px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Reduced primordial follicle activation</span></span></span></p>
<p style="text-align:justify"> </p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:92px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">(Magamage, Zengyo, Moniruzzaman, & Miyano, 2011)</span></span></span></p>
</td>
</tr>
</tbody>
</table>
<p style="text-align:justify"> </p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><em><span style="font-size:12.0pt">In vivo</span></em></strong></span></span></p>
<table align="center" cellspacing="0" class="MsoTableGrid" style="border-collapse:collapse; border:none; width:110.0%">
<tbody>
<tr>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:2px solid black; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:9.0pt">Study type</span></strong></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:2px solid black; vertical-align:top; width:68px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:9.0pt">Species</span></strong></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:2px solid black; vertical-align:top; width:71px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:9.0pt">Compound</span></strong></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:2px solid black; vertical-align:top; width:72px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:9.0pt">Dose</span></strong></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:2px solid black; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:9.0pt">Duration</span></strong></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:2px solid black; vertical-align:top; width:16px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:9.0pt">Method</span></strong></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:2px solid black; vertical-align:top; width:188px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:9.0pt">Result</span></strong></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:2px solid black; vertical-align:top; width:79px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:9.0pt">Reference</span></strong></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Fetal exposure</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:68px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Pig</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:71px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Flutamide</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:72px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">50 mg/kg body weight/day</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">7d</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:16px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Follicle counting and classification (histology)</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:188px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Reduced numbers of primary follicles and increased of primordial</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:79px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">(Knapczyk-Stwora, Grzesiak, Duda, Koziorowski, & Slomczynska, 2013)</span></span></span></p>
<p style="text-align:justify"> </p>
</td>
</tr>
<tr>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Neonatal exposure</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:68px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Pig</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:71px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Flutamide</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:72px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">50 mg/kg body weight/day</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">10d</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:16px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Immunohistochemistry (PCNA)</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:188px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Reduced number of follicles with proliferating granulosa cells</span></span></span></p>
<p style="text-align:justify"> </p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:79px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">(Knapczyk-Stwora et al., 2018)</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Neonatal exposure</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:68px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Pig</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:71px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Flutamide</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:72px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">50 mg/kg body weight/day</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">10d</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:16px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">RNAseq of whole ovary</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:188px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Altered expression of genes involved in cell proliferation</span></span></span></p>
<p style="text-align:justify"> </p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:79px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">(Knapczyk-Stwora et al., 2019)</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Adult exposure</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:68px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Rat</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:71px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Flutamide</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:72px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">One-time 100 pg</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:70px">
<p style="text-align:center"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">48h after injection</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:16px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Follicle counting and classification (histology)</span></span></span></p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:188px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">Reduced number of all stages of follicles</span></span></span></p>
<p style="text-align:justify"> </p>
</td>
<td style="border-bottom:2px solid black; border-left:none; border-right:none; border-top:none; vertical-align:top; width:79px">
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:9.0pt">(Kumari, Datta, Das, & Roy, 1978)</span></span></span></p>
</td>
</tr>
</tbody>
</table>
<p style="text-align:justify"> </p>
<p style="text-align:justify">Quality assessment of the studies was performed and can be found at: <a href="https://aopwiki.org/system/dragonfly/production/2021/03/11/3lmmehtqtn_QUALITY_ASSESSMENT_OF_STUDIES_INCLUDED_AS_EMPIRICAL_EVIDENCE.pdf">QA of Empirical Evidence</a></p>
<p style="text-align:justify"> </p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><span style="font-size:12pt">Genomic and non-genomic effects are not distinguished in the studies included in the KER analysis. Hence, it cannot be concluded that all observations are solely due to directly perturbed transcription. However, since AR transcribes genes necessary for early folliculogenesis (<em>KITLG, BMP15, HGF</em>), it is reasonable to assume that genomic mechanisms are involved.</span></span></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="color:#000000"><span style="font-size:12pt">Other uncertainties to consider: different anti-androgenic compounds have different effects on the AR (e.g. different IC<sub>50</sub>, C<sub>max</sub>); compounds that are anti-androgenic may also affect other mechanisms/modalities; downstream effects of perturbed AR transcriptional function might depend on the duration of exposure as well as the developmental stage of the follicles. In humans, effects can be inconclusive since a part of the population can have androgen-related conditions such as polycystic ovary syndrome (PCOS)<span style="color:black"><span style="font-size:11pt">(Gleicher et al., 2011)</span></span></span>.</span></span></span></p>
HighFemaleHighDuring development and at adulthoodLowHighHighHighModerateModerate<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Astapova, O., Minor, B.M.N., and Hammes, S.R. (2019). Physiological and Pathological Androgen Actions in the Ovary. Endocrinology <em>160</em>, 1166–1174. https://doi.org/10.1210/en.2019-00101.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Balasch, J., Fábregues, F., Peñarrubia, J., Carmona, F., Casamitjana, R., Creus, M., Manau, D., Casals, G., and Vanrell, J.A. (2006). Pretreatment with transdermal testosterone may improve ovarian response to gonadotrophins in poor-responder IVF patients with normal basal concentrations of FSH. Human Reproduction <em>21</em>, 1884–1893. https://doi.org/10.1093/humrep/del052.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Balen, A.H., Laven, J.S.E., Tan, S.L., and Dewailly, D. (2003). Ultrasound assessment of the polycystic ovary: International consensus definitions. Human Reproduction Update <em>9</em>, 505–514. https://doi.org/10.1093/humupd/dmg044.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Baumgarten, S.C., Convissar, S.M., Fierro, M.A., Winston, N.J., Scoccia, B., and Stocco, C. (2014). IGF1R signaling is necessary for FSH-induced activation of AKT and differentiation of human cumulus granulosa cells. Journal of Clinical Endocrinology and Metabolism <em>99</em>, 2995–3004. https://doi.org/10.1210/jc.2014-1139.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Bosdou, J.K., Venetis, C.A., Kolibianakis, E.M., Toulis, K.A., Goulis, D.G., Zepiridis, L., and Tarlatzis, B.C. (2012). The use of androgens or androgen-modulating agents in poor responders undergoing in vitro fertilization: A systematic review and meta-analysis. Human Reproduction Update <em>18</em>, 127–145. https://doi.org/10.1093/humupd/dmr051.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Caanen, M.R., Soleman, R.S., Kuijper, E.A.M., Kreukels, B.P.C., De Roo, C., Tilleman, K., De Sutter, P., Van Trotsenburg, M.A.A., Broekmans, F.J., and Lambalk, C.B. (2015). Antimüllerian hormone levels decrease in female-to-male transsexuals using testosterone as cross-sex therapy. Fertility and Sterility <em>103</em>, 1340–1345. https://doi.org/10.1016/J.FERTNSTERT.2015.02.003.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Cárdenas, H., and Pope, W.F. (2002). Androgen receptor and follicle-stimulating hormone receptor in the pig ovary during the follicular phase of the estrous cycle*. Molecular Reproduction and Development <em>62</em>, 92–98. https://doi.org/10.1002/mrd.10060.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Casson, P.R., Lindsay, M.S., Pisarska, M.D., Carson, S.A., and Buster, J.E. (2000). Dehydroepiandrosterone supplementation augments ovarian stimulation in poor responders: A case series. Human Reproduction <em>15</em>, 2129–2132. https://doi.org/10.1093/humrep/15.10.2129.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Dewailly, D., Robin, G., Peigne, M., Decanter, C., Pigny, P., and Catteau-Jonard, S. (2016). Interactions between androgens, FSH, anti-Müllerian hormone and estradiol during folliculogenesis in the human normal and polycystic ovary. Human Reproduction Update <em>22</em>, 709–724. https://doi.org/10.1093/HUMUPD/DMW027.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Erichsen, M.M., Husebye, E.S., Michelsen, T.M., Dahl, Alv.A., and Løvås, K. (2010). Sexuality and Fertility in Women with Addison’s Disease. The Journal of Clinical Endocrinology & Metabolism <em>95</em>, 4354–4360. https://doi.org/10.1210/jc.2010-0445.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Fábregues, F., Peñarrubia, J., Creus, M., Manau, D., Casals, G., Carmona, F., and Balasch, J. (2009). Transdermal testosterone may improve ovarian response to gonadotrophins in low-responder IVF patients: A randomized, clinical trial. Human Reproduction <em>24</em>, 349–359. https://doi.org/10.1093/humrep/den428.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Franks, S., and Hardy, K. (2018). Androgen action in the ovary. Frontiers in Endocrinology <em>9</em>, 452. https://doi.org/10.3389/fendo.2018.00452.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Gelmann, E.P. (2002). Molecular biology of the androgen receptor. Journal of Clinical Oncology <em>20</em>, 3001–3015. https://doi.org/10.1200/JCO.2002.10.018.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Gervásio, C.G., Bernuci, M.P., Silva-de-Sá, M.F., and Rosa-e-Silva, A.C.J. de S. (2014). The Role of Androgen Hormones in Early Follicular Development. ISRN Obstetrics and Gynecology <em>2014</em>, 1–11. https://doi.org/10.1155/2014/818010.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Gill, A., Jamnongjit, M., and Hammes, S.R. (2004). Androgens promote maturation and signaling in mouse oocytes independent of transcription: a release of inhibition model for mammalian oocyte meiosis. Molecular Endocrinology <em>18</em>, 97–104. .</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Gleicher, N., Weghofer, A., and Barad, D.H. (2011). The role of androgens in follicle maturation and ovulation induction: friend or foe of infertility treatment? Reproductive Biology & Endocrinology <em>9</em>, 116. https://doi.org/https://dx.doi.org/10.1186/1477-7827-9-116.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Gleicher, N., Kim, A., Weghofer, A., Kushnir, V.A., Shohat-Tal, A., Lazzaroni, E., Lee, H.J., and Barad, D.H. (2013). Hypoandrogenism in association with diminished functional ovarian reserve. Human Reproduction <em>28</em>, 1084–1091. https://doi.org/https://dx.doi.org/10.1093/humrep/det033.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">González-Sanz, S., Barreñada, O., Rial, E., Brieño-Enriquez, M.A., and del Mazo, J. (2020). The antiandrogenic vinclozolin induces differentiation delay of germ cells and changes in energy metabolism in 3D cultures of fetal ovaries. Scientific Reports <em>10</em>, 1–13. https://doi.org/10.1038/s41598-020-75116-3.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Guglielmo, M.C., Ricci, G., Catizone, A., Barberi, M., Galdieri, M., Stefanini, M., and Canipari, R. (2011). The effect of hepatocyte growth factor on the initial stages of mouse follicle development. Journal of Cellular Physiology <em>226</em>, 520–529. https://doi.org/10.1002/jcp.22361.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Hampton, J.H., Manikkam, M., Lubahn, D.B., Smith, M.F., and Garverick, H.A. (2004). Androgen receptor mRNA expression in the bovine ovary. Domestic Animal Endocrinology <em>27</em>, 81–88. .</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Harlow, C.R., Shaw, H.J., Hillier, S.G., and Hodges, J.K. (1988). Factors Influencing Follicle-Stimulating Hormone-Responsive Steroidogenesis in Marmoset Granulosa Cells: Effects of Androgens and the Stage of Follicular Maturity. Endocrinology <em>122</em>, 2780–2787. https://doi.org/10.1210/ENDO-122-6-2780.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Hickey, T., Chandy, A., and Norman, R.J. (2002). The androgen receptor CAG repeat polymorphism and X-chromosome inactivation in Australian Caucasian women with infertility related to polycystic ovary syndrome. Journal of Clinical Endocrinology and Metabolism <em>87</em>, 161–165. https://doi.org/10.1210/jcem.87.1.8137.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Hickey, T.E., Marrocco, D.L., Gilchrist, R.B., Norman, R.J., and Armstrong, D.T. (2004). Interactions between androgen and growth factors in granulosa cell subtypes of porcine antral follicles. Biology of Reproduction <em>71</em>, 45–52. .</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Hickey, T.E., Marrocco, D.L., Amato, F., Ritter, L.J., Norman, R.J., Gilchrist, R.B., and Armstrong, D.T. (2005). Androgens augment the mitogenic effects of oocyte-secreted factors and growth differentiation factor 9 on porcine granulosa cells. Biology of Reproduction <em>73</em>, 825–832. .</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Hillier, S.G., Tetsuka, M., and Fraser, H.M. (1997). Location and developmental regulation of androgen receptor in primate ovary. Human Reproduction <em>12</em>, 107–111. .</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Hirai, M., Hirata, S., Osada, T., Hagihara, K., and Kato, J. (1994). Androgen receptor mRNA in the rat ovary and uterus. Journal of Steroid Biochemistry & Molecular Biology <em>49</em>, 1–7. .</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Hu, Q., Hong, L., Nie, M., Wang, Q., Fang, Y., Dai, Y., Zhai, Y., Wang, S., Yin, C., and Yang, X. (2017). The effect of dehydroepiandrosterone supplementation on ovarian response is associated with androgen receptor in diminished ovarian reserve women. Journal of Ovarian Research <em>10</em>, 32. https://doi.org/https://dx.doi.org/10.1186/s13048-017-0326-3.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Jeppesen, J. V, Kristensen, S.G., Nielsen, M.E., Humaidan, P., Dal Canto, M., Fadini, R., Schmidt, K.T., Ernst, E., and Yding Andersen, C. (2012). LH-receptor gene expression in human granulosa and cumulus cells from antral and preovulatory follicles. Journal of Clinical Endocrinology & Metabolism <em>97</em>, E1524-31. https://doi.org/https://dx.doi.org/10.1210/jc.2012-1427.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Jiang, X., Teng, Y., Chen, X., Liang, N., Li, Z., Liang, D., and Wu, L. (2020). Six novel Mutation analysis of the androgen receptor gene in 17 Chinese patients with androgen insensitivity syndrome. Clinica Chimica Acta <em>506</em>, 180–186. https://doi.org/10.1016/j.cca.2020.03.036.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Juengel, J.L., Heath, D.A., Quirke, L.D., and McNatty, K.P. (2006). Oestrogen receptor alpha and beta, androgen receptor and progesterone receptor mRNA and protein localisation within the developing ovary and in small growing follicles of sheep. Reproduction <em>131</em>, 81–92. .</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Kamal, D.A.M., Ibrahim, S.F., and Mokhtar, M.H. (2020). Androgen effect on connexin expression in the mammalian female reproductive system: A systematic review. Bosnian Journal of Basic Medical Sciences <em>20</em>, 293–302. https://doi.org/10.17305/bjbms.2019.4501.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Kim, C.H., Howles, C.M., and Lee, H.A. (2011). The effect of transdermal testosterone gel pretreatment on controlled ovarian stimulation and IVF outcome in low responders. Fertility and Sterility <em>95</em>, 679–683. https://doi.org/10.1016/j.fertnstert.2010.07.1077.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Kim, C.-H., Ahn, J.-W., Moon, J.-W., Kim, S.-H., Chae, H.-D., and Kang, B.-M. (2014). Ovarian Features after 2 Weeks, 3 Weeks and 4 Weeks Transdermal Testosterone Gel Treatment and Their Associated Effect on IVF Outcomes in Poor Responders. Development & Reproduciton <em>18</em>, 145–152. https://doi.org/10.12717/dr.2014.18.3.145.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Knapczyk-Stwora, K., Grzesiak, M., Duda, M., Koziorowski, M., and Slomczynska, M. (2013). Effect of flutamide on folliculogenesis in the fetal porcine ovary--regulation by Kit ligand/c-Kit and IGF1/IGF1R systems. Animal Reproduction Science <em>142</em>, 160–167. https://doi.org/https://dx.doi.org/10.1016/j.anireprosci.2013.09.014.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Knapczyk-Stwora, K., Grzesiak, M., Ciereszko, R.E., Czaja, E., Koziorowski, M., and Slomczynska, M. (2018). The impact of sex steroid agonists and antagonists on folliculogenesis in the neonatal porcine ovary via cell proliferation and apoptosis. Theriogenology <em>113</em>, 19–26. https://doi.org/https://dx.doi.org/10.1016/j.theriogenology.2018.02.008.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Knapczyk-Stwora, K., Nynca, A., Ciereszko, R.E., Paukszto, L., Jastrzebski, J.P., Czaja, E., Witek, P., Koziorowski, M., and Slomczynska, M. (2019). Flutamide-induced alterations in transcriptional profiling of neonatal porcine ovaries. Journal of Animal Science & Biotechnology <em>10</em>, 35. https://doi.org/https://dx.doi.org/10.1186/s40104-019-0340-y.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Kumari, G.L., Datta, J.K., Das, R.P., and Roy, S. (1978). Evidence for a role of androgens in the growth and maturation of ovarian follicles in rats. Hormone Research in Paediatrics <em>9</em>, 112–120. https://doi.org/10.1159/000178903.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Laird, M., Thomson, K., Fenwick, M., Mora, J., Franks, S., and Hardy, K. (2017). Androgen Stimulates Growth of Mouse Preantral Follicles In Vitro: Interaction With Follicle-Stimulating Hormone and With Growth Factors of the TGFbeta Superfamily. Endocrinology <em>158</em>, 920–935. https://doi.org/https://dx.doi.org/10.1210/en.2016-1538.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Lebbe, M., and Woodruff, T.K. (2013). Involvement of androgens in ovarian health and disease. Molecular Human Reproduction <em>19</em>, 828–837. https://doi.org/https://dx.doi.org/10.1093/molehr/gat065.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Lebbe, M., Taylor, A.E., Visser, J.A., Kirkman-Brown, J.C., Woodruff, T.K., and Arlt, W. (2017). The Steroid Metabolome in the Isolated Ovarian Follicle and Its Response to Androgen Exposure and Antagonism. Endocrinology <em>158</em>, 1474–1485. https://doi.org/https://dx.doi.org/10.1210/en.2016-1851.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Lenie, S., and Smitz, J. (2009). Functional AR Signaling Is Evident in an In Vitro Mouse Follicle Culture Bioassay That Encompasses Most Stages of Folliculogenesis1. Biology of Reproduction <em>80</em>, 685–695. https://doi.org/10.1095/biolreprod.107.067280.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Li, Y., Wei, L.N., and Liang, X.Y. (2011). Follicle-stimulating hormone suppressed excessive production of antimullerian hormone caused by abnormally enhanced promoter activity in polycystic ovary syndrome granulosa cells. Fertility and Sterility <em>95</em>. https://doi.org/10.1016/J.FERTNSTERT.2011.03.047.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Lim, J.J., Han, C.Y., Lee, D.R., and Tsang, B.K. (2017a). Ring Finger Protein 6 Mediates Androgen-Induced Granulosa Cell Proliferation and Follicle Growth via Modulation of Androgen Receptor Signaling. Endocrinology <em>158</em>, 993–1004. https://doi.org/https://dx.doi.org/10.1210/en.2016-1866.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Lim, J.J., Lima, P.D.A., Salehi, R., Lee, D.R., and Tsang, B.K. (2017b). Regulation of androgen receptor signaling by ubiquitination during folliculogenesis and its possible dysregulation in polycystic ovarian syndrome. Scientific Reports <em>7</em>, 10272. https://doi.org/https://dx.doi.org/10.1038/s41598-017-09880-0.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Lin, L. Te, Li, C.J., and Tsui, K.H. (2021). Serum testosterone levels are positively associated with serum anti-mullerian hormone levels in infertile women. Scientific Reports 2021 11:1 <em>11</em>, 1–8. https://doi.org/10.1038/s41598-021-85915-x.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Lledo, B., Llacer, J., Turienzo, A., Ortiz, J.A., Guerrero, J., Morales, R., Ten, J., and Bernabeu, R. (2014). Androgen receptor CAG repeat length is associated with ovarian reserve but not with ovarian response. Reproductive Biomedicine Online <em>29</em>, 509–515. https://doi.org/https://dx.doi.org/10.1016/j.rbmo.2014.06.012.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Magamage, M.P.S., Zengyo, M., Moniruzzaman, M., and Miyano, T. (2011). Testosterone induces activation of porcine primordial follicles in vitro. Reproductive Medicine & Biology <em>10</em>, 21–30. https://doi.org/https://dx.doi.org/10.1007/s12522-010-0068-z.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Murray, A.A., Gosden, R.G., Allison, V., and Spears, N. (1998). Effect of androgens on the development of mouse follicles growing in vitro. Journal of Reproduction and Fertility <em>113</em>, 27–33. https://doi.org/10.1530/jrf.0.1130027.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Nagels, H.E., Rishworth, J.R., Siristatidis, C.S., and Kroon, B. (2015). Androgens (dehydroepiandrosterone or testosterone) for women undergoing assisted reproduction. The Cochrane Database of Systematic Reviews <em>2015</em>. https://doi.org/10.1002/14651858.CD009749.PUB2.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Nielsen, M.E., Rasmussen, I.A., Kristensen, S.G., Christensen, S.T., Mollgard, K., Wreford Andersen, E., Byskov, A.G., and Yding Andersen, C. (2011). In human granulosa cells from small antral follicles, androgen receptor mRNA and androgen levels in follicular fluid correlate with FSH receptor mRNA. Molecular Human Reproduction <em>17</em>, 63–70. https://doi.org/https://dx.doi.org/10.1093/molehr/gaq073.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Noventa, M., Vitagliano, A., Andrisani, A., Blaganje, M., Viganò, P., Papaelo, E., Scioscia, M., Cavallin, F., Ambrosini, G., and Cozzolino, M. (2019). Testosterone therapy for women with poor ovarian response undergoing IVF: a meta-analysis of randomized controlled trials. Journal of Assisted Reproduction and Genetics <em>36</em>, 673–683. https://doi.org/10.1007/s10815-018-1383-2.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Orisaka, M., Jiang, J.Y., Orisaka, S., Kotsuji, F., and Tsang, B.K. (2009). Growth differentiation factor 9 promotes rat preantral follicle growth by up-regulating follicular androgen biosynthesis. Endocrinology <em>150</em>, 2740–2748. https://doi.org/10.1210/en.2008-1536.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Petya Andreeva, Ivelina Oprova, Luboslava Valkova, Petya Chaveeva, Ivanka Dimova, A.S. (2020). The Benefits of Testosterone Therapy in Poor Ovarian Responders Undergoing In Vitro Fertilisation (IVF). European Medical Journal https://doi.org/10.33590/emj/20-00095.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Prizant, H., Gleicher, N., and Sen, A. (2014). Androgen actions in the ovary: Balance is key. Journal of Endocrinology <em>222</em>, 141–151. https://doi.org/10.1530/JOE-14-0296.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Rice, S., Ojha, K., Whitehead, S., and Mason, H. (2007). Stage-specific expression of androgen receptor, follicle-stimulating hormone receptor, and anti-Müllerian hormone type II receptor in single, isolated, human preantral follicles: Relevance to polycystic ovaries. Journal of Clinical Endocrinology and Metabolism <em>92</em>, 1034–1040. https://doi.org/10.1210/jc.2006-1697.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Sen, A., and Hammes, S.R. (2010). Granulosa cell-specific androgen receptors are critical regulators of ovarian development and function. Molecular Endocrinology <em>24</em>, 1393–1403. https://doi.org/10.1210/me.2010-0006.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Sen, A., Prizant, H., Light, A., Biswas, A., Hayes, E., Lee, H.J., Barad, D., Gleicher, N., and Hammes, S.R. (2014). Androgens regulate ovarian follicular development by increasing follicle stimulating hormone receptor and microRNA-125b expression. Proceedings of the National Academy of Sciences of the United States of America <em>111</em>, 3008–3013. https://doi.org/10.1073/pnas.1318978111.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Slomczynska, M., Duda, M., and Sl zak, K. (2001). The expression of androgen receptor, cytochrome P450 aromatase and FSH receptor mRNA in the porcine ovary. Folia Histochemica et Cytobiologica <em>39</em>, 9–13. .</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Steffensen, L.L., Ernst, E.H., Amoushahi, M., Ernst, E., and Lykke-Hartmann, K. (2018). Transcripts Encoding the Androgen Receptor and IGF-Related Molecules Are Differently Expressed in Human Granulosa Cells From Primordial and Primary Follicles. Frontiers in Cell & Developmental Biology <em>6</em>, 85. https://doi.org/https://dx.doi.org/10.3389/fcell.2018.00085.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Szoltys, M., and Slomczynska, M. (2000). Changes in distribution of androgen receptor during maturation of rat ovarian follicles. Experimental & Clinical Endocrinology & Diabetes <em>108</em>, 228–234. .</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Tetsuka, M., and Hillier, S.G. (1996). Androgen receptor gene expression in rat granulosa cells: the role of follicle-stimulating hormone and steroid hormones. Endocrinology <em>137</em>, 4392–4397. .</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Tetsuka, M., Whitelaw, P.F., Bremner, W.J., Millar, M.R., Smyth, C.D., and Hillier, S.G. (1995). Developmental regulation of androgen receptor in rat ovary. Journal of Endocrinology <em>145</em>, 535–543. .</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Vendola, K., Zhou, J., Wang, J., Famuyiwa, O.A., Bievre, M., and Bondy, C.A. (1999). Androgens promote oocyte insulin-like growth factor I expression and initiation of follicle development in the primate ovary. Biology of Reproduction <em>61</em>, 353–357. .</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Vendola, K.A., Zhou, J., Adesanya, O.O., Weil, S.J., and Bondy, C.A. (1998). Androgens stimulate early stages of follicular growth in the primate ovary. Journal of Clinical Investigation <em>101</em>, 2622–2629. .</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Walters, K.A., and Handelsman, D.J. (2018). Role of androgens in the ovary. Molecular and Cellular Endocrinology <em>465</em>, 36–47. https://doi.org/10.1016/j.mce.2017.06.026.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Walters, K.A., Middleton, L.J., Joseph, S.R., Hazra, R., Jimenez, M., Simanainen, U., Allan, C.M., and Handelsman, D.J. (2012). Targeted loss of androgen receptor signaling in murine granulosa cells of preantral and antral follicles causes female subfertility. Biology of Reproduction <em>87</em>. https://doi.org/10.1095/biolreprod.112.102012.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Wang, H., Andoh, K., Hagiwara, H., Xiaowei, L., Kikuchi, N., Abe, Y., Yamada, K., Fatima, R., and Mizunuma, H. (2001). Effect of adrenal and ovarian androgens on type 4 follicles unresponsive to FSH in immature mice. Endocrinology <em>142</em>, 4930–4936. .</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Weil, S., Vendola, K., Zhou, J., and Bondy, C.A. (1999). Androgen and follicle-stimulating hormone interactions in primate ovarian follicle development. Journal of Clinical Endocrinology & Metabolism <em>84</em>, 2951–2956. .</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Weil, S.J., Vendola, K., Zhou, J., Adesanya, O.O., Wang, J., Okafor, J., and Bondy, C.A. (1998). Androgen receptor gene expression in the primate ovary: cellular localization, regulation, and functional correlations. Journal of Clinical Endocrinology & Metabolism <em>83</em>, 2479–2485. .</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Xue, K., Liu, J.Y., Murphy, B.D., and Tsang, B.K. (2012). Orphan nuclear receptor NR4A1 is a negative regulator of DHT-induced rat preantral follicular growth. Molecular Endocrinology <em>26</em>, 2004–2015. https://doi.org/10.1210/me.2012-1200.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Xue, K., Kim, J.Y., Liu, J.Y., and Tsang, B.K. (2014). Insulin-like 3-induced rat preantral follicular growth is mediated by growth differentiation factor 9. Endocrinology <em>155</em>, 156–167. https://doi.org/10.1210/en.2013-1491.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Yang, M.Y., and Fortune, J.E. (2006). Testosterone stimulates the primary to secondary follicle transition in bovine follicles in vitro. Biology of Reproduction <em>75</em>, 924–932. https://doi.org/10.1095/biolreprod.106.051813.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif">Yeung, T.W.Y., Chai, J., Li, R.H.W., Lee, V.C.Y., Ho, P.C., and Ng, E.H.Y. (2014). A randomized, controlled, pilot trial on the effect of dehydroepiandrosterone on ovarian response markers, ovarian response, and in vitro fertilization outcomes in poor responders. Fertility and Sterility <em>102</em>, 108-115.e1. https://doi.org/10.1016/j.fertnstert.2014.03.044.</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"> </span></span></p>
2021-01-28T08:36:192022-04-20T11:07:04Androgen receptor (AR) antagonism leading to decreased fertility in femalesAR antagonism leading to decreased fertility<p>Eleftheria-Maria Panagiotou; Karolinska Institutet and Karolinska University Hospital, SE-14186 Stockholm, Sweden</p>
<p>Pauliina Damdimopoulou; Karolinska Institutet and Karolinska University Hospital, SE-14186 Stockholm, Sweden</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 Plan1.109<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>
adjacentModerateHighadjacentLowModerateModerate2020-05-19T16:32:182023-04-29T13:02:18