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Antagonism, Androgen receptor leads to nipple retention, increased
Key Event Relationship Overview
AOPs Referencing Relationship
|AOP Name||Adjacency||Weight of Evidence||Quantitative Understanding||Point of Contact||Author Status||OECD Status|
|Androgen receptor (AR) antagonism leading to nipple retention (NR) in male (mammalian) offspring||non-adjacent||Moderate||Low||Evgeniia Kazymova (send email)||Under development: Not open for comment. Do not cite||Under Development|
Life Stage Applicability
Key Event Relationship Description
Several chemicals can antagonize the androgen receptor (AR) in vitro, resulting in decreased AR activation. Decreased AR activation can lead to incomplete reproductive development in males, which can be expressed in several ways. One endpoint affected is areola/nipple retention (NR), which in vivo studies have shown to be linked to suppressed AR activation. NR in rat and mouse toxicity studies is considered an adverse effect (i.e., an AO).
Evidence Collection Strategy
Strategi was described by Pedersen et al (2022): A semi-systematic literature search was conducted during March 2022 in the peer-reviewed databases PubMed and Web of Science, using the search terms “(Nipple) AND (retain* OR retention) AND (androgen)” as well as “(Androgen receptor OR AR) AND (active*) AND (nipple OR areolae) AND (retain* OR retention)”. These searches resulted in 138 papers in total. Upon removal of duplicates, papers were screened according to title, abstract and ultimately full text based on pre-defined inclusion criteria. In vivo studies were included if (i) the study was carried out in mice or rats, (ii) NR in males was investigated as an endpoint, (iii) AR antagonism was the suspected mechanism of action and (iv) anti-androgenic effects of single substance exposures (i.e., not studies on chemical mixtures) were investigated. In vitro studies were included if they contained mechanistic information on AR inhibition by chemical stressors.
Evidence Supporting this KER
The biological plausibility of a link between decreased AR activation and increased NR in male rats is high. The relationship is supported by numerous studies showing that several potent AR antagonists in vitro induce NR in vivo. However, in the literature review conducted for this KER, no studies in mice were found to fulfill the inclusion criteria. The present KER is hence exclusively a description of the situation in rats, although it is believed that the link also exists in mice.
The AR is activated through binding of either testosterone or dihydrotestosterone (DHT), the latter having the highest affinity for the AR. Upon binding, the AR translocates to the target cell nucleus where it acts as a transcription factor (Albert, 2018).
NT has been shown to be more dependent on DHT-signaling, which suggests that chemicals inducing increased NR also have a higher affinity for the AR than DHT in order to outcompete DHT for AR binding, although supra-high doses of chemicals with lower AR affinity could be speculated to also outcompete T or DHT. The general principle of higher affinity, however, has been confirmed by in vitro studies (Gray et al., 2019; Hass et al., 2012; McIntyre et al., 2000).
Uncertainties and Inconsistencies
A major challenge with NR as a biomarker is the subjectivity of the measure. In juvenile rat pups, nipples are only present as areolae, i.e., dark shadows with or without a nipple bud. This means that the experience of the personnel assessing the presence and number of areolae/nipples can influence the results. Furthermore, the results are likely prone to larger variation if several assessors are used to record NR within the same study. To minimize these sources of uncertainty, assessors must be trained to recognize areolae and not look for fully developed nipples. Moreover, the number of assessors should be limited to one or two, and they should always be blinded to exposure groups.
Another factor that may affect NR results is the age of the rat pups at the time of assessment. OECD guidelines have standardized the time for measuring occurrence of NR to be optimal at PD 12 or 13, when they are visible in female littermates (OECD, 2013). However, assessment of permanent NR is not included in any international guidelines. Hence, if NR is measured in older offspring, the time of measurement is not consistent between studies and varies between PD 20 and PD 100. Thus, conclusions on whether NR is permanent or not may differ based on study design. This distinction between a transient and a permanent effect is important from a regulatory perspective, since only a permanent effect will be categorized as a malformation according to OECD guidance document 43 (OECD, 2008).
Known modulating factors
No response-response relationship has been identified.
NR manifest in juvenile male rat pup offspring in response to reduced androgen signaling, e.g. resulting from exposure to an anti-androgenic chemical stressor during fetal development. Developmental sensitivity during fetal development is highest during the so-called male masculinization programming window (MPW) which in rats is between gestational day (GD) 15 and 19 (Welsh et al., 2008).
A study in which pregnant rat dams were exposed to the AR antagonist vinclozolin for two-day periods during gestation showed that GD 16–17 was the most sensitive period for increased NR in male offspring (Wolf et al., 2000). A similar study using di-n-butyl phthalate (reduces testosterone levels) also showed that GD 16–17 was the most sensitive period for increased NR in male rats (Carruthers & Foster, 2005). However, to determine if other chemical stressors also have the highest antagonistic potential towards the AR during GD 16-17, further studies with a similar design would be informative.
NR can only be recorded when pups are old enough to display them, yet before excessive fur has developed. Hence, the most accurate results can be obtained from assessing the number of nipples on PD 12–14 depending on rat strain and the time of female littermates displaying nipples (OECD, 2013).
Known Feedforward/Feedback loops influencing this KER
No feedback loops that could influence the KER have been identified.
Domain of Applicability
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