Androgen Bioavailability Research Articles

Discovery of a Series of Orally Bioavailable Androgen Receptor Degraders for the Treatment of Prostate Cancer.

Androgen receptor (AR) signaling plays a key role in the progression of prostate cancer. This study describes the discovery and optimization of a novel series of AR PROTAC degraders that recruit the Cereblon (CRBN) E3 ligase. Having identified a series of AR ligands based on 4-(4-phenyl-1-piperidyl)-2-(trifluoromethyl)benzonitrile, our PROTAC optimization strategy focused on linker connectivity and CRBN ligand SAR to deliver potent degradation of AR in LNCaP cells. This work culminated in compounds 11 and 16 which demonstrated good rodent oral bioavailability. Subsequent SAR around the AR binding region brought in an additional desirable feature, degradation of the important treatment resistance mutation L702H. Compound 22 (AZ'3137) possessed an attractive profile showing degradation of AR and L702H mutant AR with good oral bioavailability across species. The compound also inhibited AR signaling in vitro and tumor growth in vivo in a mouse prostate cancer xenograft model.

RISING STARS: Androgens and immune cell function.

Androgens can modulate immune cell function and may contribute to differences in the prevalence and severity of common inflammatory conditions. Although most immune cells are androgen targets, our understanding of how changes in androgen bioavailability can affect immune responses is incomplete. Androgens alter immune cell composition, phenotype, and activation by modulating the expression and secretion of inflammatory mediators or by altering the development and maturation of immune cell precursors. Androgens are generally associated with having suppressive effects on the immune system, but their impacts are cell and tissue context-dependent and can be highly nuanced even within immune cell subsets. In response to androgens, innate immune cells such as neutrophils, monocytes, and macrophages increase the production of the anti-inflammatory cytokine IL-10 and decrease nitric oxide production. Androgens promote the differentiation of T cell subsets and reduce the production of inflammatory mediators, such as IFNG, IL-4 and IL-5. Additionally, androgens/androgen receptor can promote the maturation of B cells. Thus, androgens can be considered as immunomodulatory agents, but further work is required to understand the precise molecular pathways that are regulated at the intersection between endocrine and inflammatory signals. This narrative review focusses on summarising our current understanding of how androgens can alter immune cell function and how this might affect inflammatory responses in health and disease.

(100) The Effect of Oral Contraceptive Pill Type on Vulvovaginal Atrophy

Abstract Introduction Oral contraceptive pills (OCPs), which are used by over a quarter of reproductive-age women in the United States (Daniels et al., 2015; Jones et al., 2013), contain either both ethinylestradiol and a synthetic progestin, or solely a synthetic progestin. Through progestin-induced hepatic increases in sex hormone binding globulin (SHBG), OCPs have been found to reduce the number of bioavailable androgens within women’s bodies (Zimmerman et al., 2014). Given that androgens are essential for the maintenance of healthy vaginal tissue (I. Goldstein, 2009), reduced androgen levels may adversely impact the vaginal epithelium in OCP users. While OCPs are indeed associated with symptoms of vulvovaginal atrophy (VVA), such as vaginal pain and vaginal dryness (Battaglia et al., 2012), little is known about how these symptoms may vary based on the androgenicity of OCPs. Objective The aim of the present study was to compare rates of VVA among three groups of women: (1) women using OCPs containing low doses (i.e., ≤25 μg) of ethinylestradiol coupled with an androgenic progestin, (2) women using OCPs containing low doses (i.e., ≤25 μg) of ethinylestradiol coupled with an antiandrogenic progestin, and (3) naturally-cycling women (i.e., women with no history of oral contraceptive pill use). Methods Participants rated the severity of the following symptoms of VVA on a 4-point Likert-type scale ranging from 0 (mild) to 3 (severe): vaginal dryness, vaginal and/or vulvar irritation/itching, dysuria, and vaginal pain associated with sexual activity. Participants also dichotomously rated the presence or absence of vaginal bleeding associated with sexual activity. Results Women taking androgenic OCPs (n = 50) were significantly more likely to report vaginal bleeding associated with sexual activity than were women in the control group (n = 59), χ2(2, 130) = 9.759, p = .007. One participant (1.69%) in the control reported vaginal bleeding, compared to ten women (20%) in the androgenic group. Three participants (14.28%) in the antiandrogenic group endorsed vaginal bleeding, however this result was not significant. A MANOVA indicated no significant between-group differences for any of the remaining VVA items (i.e., dryness, itching/irritation, dysuria, or pain), F(2, 250) = 1.098, p = .365, and an ANOVA confirmed there were no differences between groups in the total VVA score, F(2) = 2.093, p = .127. Conclusions There were elevated rates of vaginal bleeding among OCP users compared to non-OCP users in this sample. This finding could be attributable a range of factors which have previously linked OCP use to vaginal bleeding, such as thinning of the vaginal tissue (Kagan & Rivera, 2018), cervical ectopy (Patil & Sharma, 2017), and thinning of the endometrium (Bright et al., 2011). Disclosure No

Impact of various dietary interventions on the reduction of symptoms in polycystic ovary syndrome (PCOS)

Introduction and purpose. Polycystic ovary syndrome (PCOS) is a common endocrine and metabolic disorder diagnosed in women. Symptoms include menstrual disorders, infertility, acne, seborrhea, hirsutism and carbohydrate metabolism disorders. Treatment is based on the reduction of symptoms, as it is not possible to completely cure the disease. To achieve the optimal therapeutic effect, it is advisable to change eating habits. The aim of the study is to summarize the effectiveness of the mediterranean diet, ketogenic diet, lactose-free diet and supplementation in reducing PCOS symptoms. State of knowledge. The mediterranean diet, which is includes large amounts of fish rich in omega-3 fatty acids, leads to a reduction in the severity of PCOS symptoms by reducing the bioavailability of androgens and lowering the concentration of triglycerides and glucose in the blood serum. A high-fat ketogenic diet reduces postprandial insulin release, improving insulin sensitivity. In addition, it causes a significant reduction in body fat, which in turn reduces the level of free testosterone and improves the LH/FSH ratio. Milk proteins have a strong effect on the secretion of lactose and incretin, however, their effect on insulin resistance is still unclear. Conclusions. The most effective nutritional model is the mediterranean diet, as it reduces the risk of other diseases such as hypertension, diabetes and osteoarthritis. The ketogenic diet remains an alternative solution, but it should be used for a limited time. The least effective intervention turns out to be a lactose-free diet. To obtain optimal therapeutic effects, the important role of PUFA, inositol and probiotic supplementation should not be forgotten.

Replication study of THADA rs13429458 variant with PCOS susceptibility and its related traits in Indian women

Aim The aetiopathogenesis of the multigenic multifactorial endocrinopathy, polycystic ovary syndrome (PCOS) has been explored using linkage, candidate gene and genome-wide association studies. Contradictory reports of replication studies attributed to phenotypic, ethnic and geographic variations are available. In this study, we investigated the association of Han Chinese GWAS polymorphism (rs13429458) in thyroid adenoma-associated gene (THADA) with PCOS susceptibility and its related traits in Indian women. Methods We genotyped rs13429458 of THADA by direct sequencing and investigated its association with PCOS and its related traits in controls (N = 150) and PCOS women (N = 348). All women were extensively phenotyped in terms of anthropometric, hormonal and metabolic parameters. Association of polymorphism with PCOS risk and its related traits was carried out by regression analysis. Results Genotypic and allele frequencies for rs13429458 were not different between controls and PCOS. Women with PCOS carrying variant allele showed significantly reduced fasting glucose levels, and decreased free and bioavailable testosterone and free androgen index. Conclusion To our knowledge, this is the first study to demonstrate that although this polymorphism does not alter PCOS susceptibility, it favorably impacts glucose metabolism and hyperandrogenism in Indian women with PCOS only. This study highlights that genetic predisposition markers for PCOS may differ with ethnicity and phenotypic variations.

Menstrual Disorders and Androgen-Related Traits in Young Women with Type 1 Diabetes Mellitus: a Clinical Study

To investigate possible causes of menstrual disorders and androgen-related traits in young women with type 1 diabetes mellitus (T1DM). Fifty-three women with T1DM (duration 8.0±5.6 years), 41 women with (polycystic ovary syndrome) PCOS, and 51 controls matched for age (19.4±4.3 years vs. 21.2±2.7 years vs. 20.8±3.1 years; P>.05) and body mass index (BMI) (22.2±2.7 kg/m2 vs. 21.9±2.0 kg/m2 vs. 21.4±1.9 kg/m2; P>.05) were prospectively recruited. Two women (3.8%) in the T1DM group had not experienced menarche (at 15.5 and 16.6 years); of the rest, 23.5% had oligomenorrhea, 32.1% hirsutism, and 45.3% had acne. The age at menarche was delayed in the T1DM group compared to controls (12.7±1.3 vs. 12.0±1.0 years; P = .004), while no difference was observed with the polycystic ovary syndrome (PCOS) group (12.4±1.2 years). There were no differences in total testosterone (0.43±0.14 ng/mL vs. 0.39±0.14 ng/mL; P>.05), dehydroepiandrosterone sulfate (DHEA-S) (269 ± 112 μg/dL vs. 238 ± 106 μg/dL; P>.05) or Δ4-androstenedione (2.4±1.3 ng/mL vs. 1.9±0.5 ng/mL; P>.05) concentrations between T1DM and controls. However, patients with T1DM had lower sex hormone binding globulin (SHBG) concentrations than controls (61 ± 17 nmol/L vs. 83 ± 18.1 nmol/L; P = .001), which were even lower in the PCOS group (39.5±12.9 nmol/L; P = .001 compared with T1DM). The free androgen index (FAI) was higher in the PCOS group compared with both other groups (T1DM vs. PCOS vs. controls: 2.53±0.54 vs. 7.88±1.21 vs. 1.6 ± 0.68; P<.001). FAI was higher in patients with T1DM compared to controls as well (P = .038). There was no difference in DHEA-S concentrations between T1DM and PCOS patients (269 ± 112 μg/dL vs. 297 ± 100 μg/dL; P>.05). Menstrual disorders and androgen-related traits in young women with T1DM may be attributed to an increase in androgen bioavailability due to decreased SHBG concentrations.

020 Oral Contraceptive use Influences Subjective and Physiological Sexual Arousal

Genital sexual arousal, which consists of genital vasocongestion and lubrication, is critical to healthy sexual function in women and closely linked with hormone function. Oral hormonal contraceptive pills (OCPs), which are used by over a quarter of reproductive-age women in the United States, tend to reduce the number of bioavailable androgens and have been associated with decrements in self-reported arousal and vaginal lubrication. However, empirical examinations of the possible physiological effects of OCPs on women’s sexual arousal response are sparse.

SPERMIDINE DOES NOT INFLUENCE LIMB MUSCLE MASS FOLLOWING ANDROGEN DEPLETION

Skeletal muscle atrophy increases the risk of morbidity and mortality during various pathological conditions. In males, a decrease in the production and/or bioavailability of androgens (termed hypogonadism) directly contributes to muscle atrophy during such conditions. While it is known that androgens prevent muscle atrophy, the mechanism(s) by which androgens mediate this effect are largely undefined. To identify potential androgen sensitive pathways regulating limb muscle mass, we subjected total RNA from the tibialis anterior (TA) from castrated mice and total RNA from the TA of sham mice to microarray analysis. This approach showed that androgen depletion significant altered the expression of genes involved in the polyamine metabolic pathway. The polyamines, spermidine (Spd), spermine (Spm) and their diamine precursor putrescine, are naturally occurring, polycationic alkylamines that are absolutely required for mammalian cell growth, differentiation, and survival. Accordingly, we confirmed that androgen depletion reduced muscle expression of ornithine decarboxylase (ODC1), S‐adenosylmethionine decarboxylase (AMD1), spermine oxidase (SMOX), and spermidine/spermine N‐1acetyltrasferase (SAT1), which are genes involved with polyamine biosynthesis (ODC1, AMD1) and catabolism (SMOX, SAT1). Changes in the expression of these polyamine metabolic genes following androgen depletion coincided with a preferential decrease in Spd concentration in the TA, and this reduction was strongly related to the degree of muscle atrophy induced by the androgen depletion. To assess the role of Spd in the regulation of limb muscle mass, we treated previously castrated mice with Spd in the drinking water for 8 weeks. Even though Spd treatment completely prevented the decrease in muscle Spd concentrations following androgen depletion, mass of the TA or other limb skeletal muscles was not affected. Moreover, treating previously castrated mice with nandrolone decanoate increased muscle mass and restored expression of the polyamine metabolic genes without restoring Spd values. Thus, the decrease in Spd concentration in the limb muscle following androgen depletion likely mediates processes that are independent of muscle mass regulation.Support or Funding InformationNIH Grant CA204345 to (RAC) Institute for Successful Longevity to (BSG)

Expression of Genes that Comprise the Core Molecular Clock are Altered in the Atrophied Skeletal Muscle by Androgen Deprivation

Skeletal muscle atrophy increases the risk of morbidity and mortality during various pathological conditions. In males, a decrease in the production and/or bioavailability of androgens (termed hypogonadism) directly contributes to muscle atrophy during various pathological conditions. While it is known that androgens prevent muscle atrophy, the mechanism(s) by which androgens mediate this effect are largely undefined. Our laboratory previously showed that mitochondrial turnover is enhanced in the tibialis anterior (TA) muscle of mice by androgen deprivation induced by castration surgery, and the magnitude of turnover was related to the degree of muscle atrophy. These data suggest that potentially dysfunctional mitochondria contribute to the muscle atrophy observed following androgen deprivation. To gain a better understanding of that factors that might contribute to changes in mitochondrial quality control during androgen deprived conditions, we subjected total RNA from the TA of sham and castrated mice to microarray analysis. This unbiased approach identified significant changes in expression of genes that comprise the core molecular clock. qRT‐PCR confirmed that expression of Brain and Muscle Arntl 1 (Bmal1) was decreased, while expression of Period 1, Period 2, and Period 3 (Per1, 2, &amp; 3) were increased in the TA of castrated mice. When measured across a diurnal cycle, the change in expression of Bmal1, Per1, and Per2 exhibited reduced amplitude under androgen‐deprived conditions. Interestingly, strong relationships were observed between the castration‐mediated changes of core clock components and the measures of mitochondrial turnover. Specifically, Bmal1 expression was directly related to BCL2/adenovirus E1B 19 kDA protein‐interacting protein 3 (BNIP3) protein content (R2 = 0.88), and the expression of core clock components were also directly related to the content of various mitochondrial proteins. Expression of core clock components were also related to the autophagy marker, p62, and the mass of the TA. Ex post facto analysis of the microarray also identified changes in genes regulating polyamine biosynthesis. As polyamines are known to alter core clock function, we determined whether androgen deprivation altered polyamine content. While castration did not alter Spermine content, there was a significant reduction in the content of Spermidine in the TA of castrated mice. Overall, these data suggest that reduced Spermidine concentrations may contribute to alterations in the core molecular clock in the skeletal muscle under androgen‐deprived conditions, which may in turn contribute to reduced mitochondrial quality control and subsequent muscle atrophySupport or Funding InformationNIH‐CA204345 (RAC) and Institute for Successful Longevity (BSG)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Abstract 5236: ARV-110: An androgen receptor PROTAC degrader for prostate cancer

Abstract The Androgen Receptor (AR) remains the principal driver of castration-resistant prostate cancer during the transition from a localized to metastatic disease. Most patients initially respond to inhibitors of the AR pathway, but the response is often short-lived. The majority of patients progressing on enzalutamide or abiraterone exhibit genetic alterations in the AR locus, either in the form of amplifications or point mutations in the AR gene. Given these mechanisms of resistance, our goal is to eliminate the AR protein using the PROteolysis TArgeting Chimera (PROTAC) technology. Here we report an orally bioavailable small molecule ARV-110 that leads to ubiquitination and degradation of AR. ARV-110 completely degrades AR in all cell lines tested, with an observed 50% degradation concentration (DC50) &amp;lt; 1 nM. PROTAC-mediated AR degradation suppresses the expression of the AR-target genes PSA and FKBP5, inhibits AR-dependent cell proliferation, and induces potent apoptosis in VCaP cells. ARV-110 degrades clinically relevant mutant AR proteins and retains activity in a high androgen environment. In mouse xenograft studies, greater than 90% AR degradation is observed at a 1 mg/kg PO QD dose. Significant inhibition of tumor growth and AR signaling can be achieved in both an intact and castrate setting. Further, ARV-110 demonstrates in vivo efficacy and reduction of oncogenic Erg protein in a long term, castrate, enzalutamide-resistant VCaP tumor model. DMPK and exploratory toxicology studies show robust oral, dose proportional drug exposure in rodent and non-rodent species. In summary, we report preclinical data on ARV-110, an orally bioavailable androgen receptor PROTAC degrader that demonstrates efficacy in enzalutamide-resistant prostate cancer. Citation Format: Taavi Neklesa, Lawrence B. Snyder, Ryan R. Willard, Nicholas Vitale, Kanak Raina, Jennifer Pizzano, Deborah Gordon, Mark Bookbinder, Jennifer Macaluso, Hanqing Dong, Zheng Liu, Caterina Ferraro, Gan Wang, Jing Wang, Craig M. Crews, John Houston, Andrew P. Crew, Ian Taylor. ARV-110: An androgen receptor PROTAC degrader for prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5236.

Vitamin D is associated with bioavailability of androgens in eumenorrheic women with prior pregnancy loss

Prior studies have reported mixed results regarding relationships between vitamin D, androgens, and sex hormone-binding globulin in patients with polycystic ovary syndrome. However, less is known regarding these associations in eumenorrheic, premenopausal women. Our objective was to study the relationships betweenserum vitamin D and androgen biomarkers in eumenorrheic women with a history of pregnancy loss who were attempting pregnancy. This was an analysis of a cohort of 1191 participants from the Effects of Aspirin in Gestation and Reproduction trial (2006-2012). Participants were attempting to conceive, aged 18-40 years, with 1-2 documented prior pregnancy losses and no history ofinfertility, and recruited from 4 academic medical centers in the United States. Serum vitamin D (25-hydroxyvitamin D) and hormone concentrations were measured at baseline. Vitamin D concentration was negatively associated with free androgen index (percentage change [95% confidence interval, -5% (-8% to -2%)] per 10 ng/mL increase) and positively associated with sex hormone-binding globulin (95% confidence interval, 4% [2-7%]), although not with total testosterone, free testosterone, or dehydroepiandrosterone sulfate after adjusting for age, body mass index, smoking status, race, income, education, physical activity, and season of blood draw. Overall, vitamin D was associated with sex hormone-binding globulin and free androgen index in eumenorrheic women with prior pregnancy loss, suggesting that vitamin D may play a role in the bioavailability of androgens in eumenorrheic women. We are limited in making assessments regarding directionality, given the cross-sectional nature of our study.

The role of androgens in the regulation of muscle oxidative capacity following aerobic exercise training.

Reduced production or bioavailability of androgens, termed hypogonadism, occurs in a variety of pathological conditions. While androgens target numerous tissues throughout the body, hypogonadism specifically reduces the ability of skeletal muscle to produce adenosine triphosphate aerobically, i.e., muscle oxidative capacity. This has important implications for overall health as muscle oxidative capacity impacts a number of metabolic processes. Although androgen replacement therapy is effective at restoring muscle oxidative capacity in hypogonadal individuals, this is not a viable therapeutic option for all who are experiencing hypogonadism. While aerobic exercise may be a viable alternative to increase muscle oxidative capacity, it is unknown whether androgen depletion affects this adaptation. To determine this, sham and castrated mice were randomized to remain sedentary or undergo 8 weeks of aerobic treadmill exercise training. All mice were fasted overnight prior to sacrifice. Though exercise increased markers of muscle oxidative capacity independent of castration (cytochrome c oxidase subunit IV and cytochrome c), these measures were lower in castrated mice. This reduction was not due to a difference in peroxisome proliferator activated receptor gamma coactivator 1 alpha protein content, as expression was increased to a similar absolute value in sham and castrated animals following exercise training. However, markers of BCL2/Adenovirus E1B 19 kDa Interacting Protein 3 (BNIP3)-mediated mitophagy were increased by castration independent of exercise. Together, these data show that exercise training can increase markers of muscle oxidative capacity following androgen depletion. However, these values are reduced by androgen depletion likely due in part to elevated BNIP3-mediated mitophagy.

It is time for new male contraceptives!

It is time for new male contraceptives!

Relationship Between Aerobic Fitness, the Serum IGF-1 Profiles of Healthy Young Adult African American Males, and Growth of Prostate Cancer Cells.

The growth of prostate tumors is mediated by the bioavailability of androgens and insulin-like growth factors. This study tested the hypothesis that healthy young adult African American men exhibiting low aerobic capacity (fitness) have serum insulin-like growth Factor-1 (IGF-1) and testosterone levels that promote growth of prostate cancer cells. A cross-sectional data research design was used to study groups of 18- to 26-year-old healthy men exhibiting low and moderate aerobic fitness, based on their peak oxygen consumption (VO2peak). The individual serum levels of IGF-1, IGF-1 binding protein-3 (IGFBP-3), and testosterone were measured. In vitro growth of androgen-dependent LNCaP prostate tumor cells was measured after incubation in culture medium fortified with each subject's serum. Aerobic capacity was significantly greater in the moderate-fitness group than in the low-fitness group without an intergroup difference in body mass index. The serum IGF-1 concentration was significantly higher in the low-fitness group in the absence of an intergroup difference in serum testosterone. The serum IGFBP-3 concentration was significantly lower in the low-fitness group. Prostate tumor cell growth was significantly greater in the cultures incubated in media containing the sera of the low-fitness group than in the sera of the moderate-fitness group. These findings suggest that moderate aerobic fitness in young adults may decrease the circulating levels of free IGF-1 and lower the potential to support growth of prostate cancer cells.

Epigenetic regulation of steroid inactivating UDP-glucuronosyltransferases by microRNAs in prostate cancer

Androgens play a central role in prostate cancer progression. Systemic and local androgen bioavailability is controlled by UDP-glucuronosyltransferases conjugating enzymes (UGT), namely UGT2B15, UGT2B17 and UGT2B28. Reporter vector assays in HEK293 cells initially validated in silico-predicted regulatory potential of candidate miRNAs to target UGT transcripts, including miR-376c, miR-409 and miR-494 for UGT2B17, miR-331-5p and miR-376c for UGT2B15 while none were efficient for UGT2B28. miR-376c was shown as the most effective to downregulate UGT2B15 and UGT2B17 through interactions with a site conserved in both UGTs. Ectopic miR-376c expression in prostate cancer cells significantly reduced UGT2B15 and UGT2B17 expression (>32%; P<0.005) with a consequent decrease in dihydrotestosterone glucuronidation (−37%; P<0.001). Consistent with reduced androgen inactivation, ectopic expression of miR-376c changed expression of androgen responsive genes and enhanced cell proliferation with no effect on androgen receptor levels. Sustaining a role of miR-376c in the regulation of androgen-inactivating UGTs, its expression was significantly downregulated in prostatic tumors and further reduced in metastases (P<0.0001), whereas the opposite was observed for UGT2B15/17 (P=0.031). In high-grade tumors (Gleason ≥8), UGT2B15/17 and miR-376c were inversely correlated (r=−0.557; P=0.048) with also a significant relationship in metastases (r=−0.747; P=0.003). In line with a modification in androgen bioavailability, PSA mRNA levels were also negatively correlated to those of UGT2B15/17 (r=−0.573; P=0.01) but positively linked to levels of miR-376c (r=0.577; P=0.039). This study reveals that the androgen-inactivating UGT2B15 and UGT2B17 genes are direct targets of miR-376c and thus may influence steroid metabolism during prostate cancer progression.

Abstract 3448: Overexpression of the steroid inactivating UGT2B28 enzyme is associated with high circulating androgens, tumor aggressiveness and adverse prostate cancer outcome

Abstract Androgen inactivation occurs mainly through the glucuronidation conjugative reaction mediated by uridine diphospho-glucuronosyltransferase enzymes, namely UGT2B28. This metabolic pathway controls systemic and local androgen bioavailability along with other biotransformation and transport pathways. In this study, we examined the relationship between UGT2B28 protein expression, circulating steroid levels and prostate cancer phenotype. To study distribution of the UGT2B28 protein in prostatic tissues, we used a specific antipeptide antibody displaying no cross-reactivity against highly identical UGT2B family members. Findings revealed that UGT2B28 cytoplasmic staining of secretory cells was distinctive of early stage tumorigenesis (i.e. prostatic intraepithelial neoplasia) and cancer cells with a variable nuclear staining, whereas normal prostate presented only nuclear enzyme expression. Using tissue microarray analysis, we analyzed 239 localized prostate cancer cases that underwent radical prostatectomy. Compared to patients with prostate tumors displaying absent or weak UGT2B28 staining, those with tumors presenting strong UGT2B28 nuclear expression had significantly lower PSA levels at diagnosis (20%; p = 0.043). UGT2B28 expression in the cytoplasm was also correlated with Gleason scores (p≤0.03) and nodal invasion status (p = 0.024). After adjustments for known prognostic markers including PSA, Gleason score, margin and nodal status, tumor size, smoking and germline UGT2B28 deletion status, overexpression of the UGT2B28 protein was an adverse and independent prognostic factor associated with disease recurrence and/or death (HR = 2.82; 95%CI, 1.09-7.36; p = 0.033). Remarkably, prostatic overexpression of UGT2B28 concurs with 30% higher circulating levels of testosterone (p = 0.004) and dihydrotestosterone (p = 0.002), measured by mass spectrometry. Our observations demonstrate that an increased expression of the UGT2B28 enzyme is associated with circulating steroid hormone levels, disease aggressiveness and risk of cancer progression. Citation Format: Anaïs Belledant, Hélène Hovington, Hervé Brisson, Bernard Têtu, Yves Fradet, Louis Lacombe, Patrick Caron, Chantal Guillemette, Eric Lévesque. Overexpression of the steroid inactivating UGT2B28 enzyme is associated with high circulating androgens, tumor aggressiveness and adverse prostate cancer outcome. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3448. doi:10.1158/1538-7445.AM2015-3448

The UGT2B28 Sex-steroid Inactivation Pathway Is a Regulator of Steroidogenesis and Modifies the Risk of Prostate Cancer Progression

BackgroundAndrogen inactivation occurs mainly through the glucuronidation conjugative reaction mediated by UDP-glucuronosyltransferases (UGTs). This metabolic process is involved in the control of systemic and local androgen bioavailability. ObjectiveTo examine the relationship among expression of the androgen-inactivating UGT2B28 enzyme, circulating steroid hormone levels, and clinical phenotype in prostate cancer (PCa). Design, setting, and participantsWe conducted an analysis of a high-density prostate tumor tissue microarray consisting of 239 localized PCa cases. The study of 51 additional PCa patients with no copies of UDP glucuronosyltransferase 2B subfamily, polypeptide B28 (UGT2B28) in their genomes was performed to confirm the importance of the enzyme on circulating hormone levels. Outcome measurements and statistical analysisSteroid hormones were measured by mass spectrometry. Multivariate Cox proportional hazard models assessed the influence of UGT2B28 on progression, and general linear model regression evaluated variations in hormone levels. Results and limitationsTumor overexpression of UGT2B28 was associated with lower prostate-specific antigen levels at diagnosis, higher Gleason scores, margin and nodal invasion status, and it was shown to be an independent prognostic factor associated with progression. Enzyme overexpression correlated with 30% higher circulating levels of testosterone (T) and dihydrotestosterone (DHT). Patients with no copies of UGT2B28 in their genomes have lower levels of T (19%), DHT (17%), its glucuronide metabolites (18–38%), and enhanced levels of the adrenal precursor androstenedione (36%). ConclusionsThe UGT2B28 steroid-inactivating pathway modifies circulating T and DHT levels, and UGT2B28 overexpression is associated with high-grade PCa. Our work has uncovered the role of UGT2B28 as a regulator of steroidogenesis and underscores the interconnectivity among the steroid-inactivation capacity of cancer cells, hormone levels, disease characteristics, and the risk of cancer progression. Patient summaryThe androgen-inactivating UGT2B28 enzyme influences hormone levels, clinical and pathologic factors, and the risk of cancer progression.

Protective effect of alpha glucosyl hesperidin (G-hesperidin) on chronic vanadium induced testicular toxicity and sperm nuclear DNA damage in male Sprague Dawley rats.

The study was conducted to evaluate the vanadium-induced testicular toxicity and its effect on sperm parameters, sperm nuclear DNA damage and histological alterations in Sprague Dawley rats and to assess the protective effect of G-hesperidin against this damage. Treatment of rats with vanadium at a dose of 1mgkgbw(-1) for 90days resulted in significant reduction in serum testosterone levels, sperm count and motility. Further, a parallel increase in abnormal sperm morphology and adverse histopathological changes in testis was also associated with vanadium administration when compared to normal control. Moreover, sperm chromatin dispersion assay revealed that vanadium induces sperm nuclear DNA fragmentation. A marked increase in testicular malondialdehyde levels and decreased activity of antioxidant enzymes such as superoxide dismutase and catalase indicates vanadium-induced oxidative stress. Co-administration of G-hesperidin at a dose of 25 and 50mgkgbw(-1) significantly attenuated the sperm parameters and histological changes by restoring the antioxidant levels in rat testis. These results suggested that vanadium exposure caused reduced bioavailability of androgens to the tissue and increased free radical formation, thereby causing structural and functional changes in spermatozoa. G-hesperidin exhibited antioxidant effect by protecting the rat testis against vanadium-induced oxidative damage, further ensures antioxidant potential of bioflavonoids.

Childhood Obesity and Its Impact on the Development of Adolescent PCOS

Obesity exacerbates the reproductive and metabolic manifestations of polycystic ovary syndrome (PCOS). The symptoms of PCOS often begin in adolescence, and the rising prevalence of peripubertal obesity has prompted concern that the prevalence and severity of adolescent PCOS is increasing in parallel. Recent data have disclosed a high prevalence of hyperandrogenemia among peripubertal adolescents with obesity, suggesting that such girls are indeed at risk for developing PCOS. Obesity may impact the risk of PCOS via insulin resistance and compensatory hyperinsulinemia, which augments ovarian/adrenal androgen production and suppresses sex hormone-binding globulin (SHBG), thereby increasing androgen bioavailability. Altered luteinizing hormone (LH) secretion plays an important role in the pathophysiology of PCOS, and although obesity is generally associated with relative reductions of LH, higher LH appears to be the best predictor of increased free testosterone among peripubertal girls with obesity. Other potential mechanisms of obesity-associated hyperandrogenemia include enhanced androgen production in an expanded fat mass and potential effects of abnormal adipokine/cytokine levels. Adolescents with PCOS are at risk for comorbidities such as metabolic syndrome and impaired glucose tolerance, and concomitant obesity compounds these risks. For all of these reasons, weight loss represents an important therapeutic target in obese adolescents with PCOS.

Relationship of Adiponectin with Sex Hormone Levels in Adolescents

Background/Aims: Adiponectin is an adipose tissue-derived adipocytokine which, starting at puberty, is present in lower levels in males than in females. In adults, a relationship between adiponectin levels and sexual hormones has been suggested, but this association remains unclear in children. Our study aimed to analyze the relationship of adiponectin with dehydroepiandrosterone sulfate, testosterone, estradiol, and sex hormone-binding globulin (SHBG) in adolescents. Methods: The population-based sample included 785 healthy 12- to 16-year-old children. Testosterone, estradiol, and dehydroepiandrosterone sulfate levels were assessed by radioimmunoassay. SHBG was measured by immunoradiometric assay. Adiponectin concentrations were measured by ELISA. Results: Adiponectin levels were positively correlated with SHBG and negatively correlated with the free androgen index independently of body mass index (BMI) and fat mass. No significant correlations were observed between adiponectin and total testosterone or estradiol. Multiple linear regression analysis showed that, after adjustment for BMI, SHBG appears as the primary predictor of adiponectin levels in both sexes, accounting for 5.7% of the variation in adiponectin levels in boys and 7.5% in girls. Conclusion: Adiponectin is related to free androgen index and SHBG levels in adolescents after adjusting for BMI and fat mass, thus suggesting an association between adiponectin levels and androgen bioavailability that would explain the sex-based differences in adiponectin levels through life.