Antiandrogenic Effects Research Articles

Estrogenic, androgenic, and genotoxic activities of zearalenone and deoxynivalenol in in vitro bioassays including exogenous metabolic activation

Zearalenone (ZEN) and deoxynivalenol (DON) and their derivatives are well-known mycotoxins, which can occur not only in crops but also in water bodies, including drinking water sources. In vitro bioassays can be used to detect biological effects of hazardous compounds in water. To this, when studying biological effects and toxicity in vitro, metabolism is important to consider. In this study, ZEN, α-zearalenol (α-ZEL), DON, 3-acetyl DON, and 15-acetyl DON were evaluated in vitro for hormone receptor–mediated effects (estrogen receptor [ER] and androgen receptor [AR]) and genotoxicity (micronucleus assay) in the presence of an exogenous metabolic activation system (MAS). The ER bioassay proved to be a highly sensitive method to detect low concentrations of the ZEN compounds (EC10 values of 31.4 pM for ZEN, 3.59 pM for α-ZEL) in aqueous solutions. In the presence of the MAS, reduced estrogenic effects were observed for both ZEN compounds (EC10 values of 6.47 × 103 pM for ZEN, 1.55 × 102 pM for α-ZEL). Of the DON compounds, only 3-acetyl DON was estrogenic (EC10 of 0.31 µM), and the effect was removed in the presence of the MAS. Anti-androgenic effects of the ZEN compounds and androgenic effects of the DON compounds were detected in the micromolar range. No induction of genotoxicity was detected for ZEN or DON in the presence of the MAS. Our study highlighted that inclusion of exogenous MAS is a useful tool to detect biological effects of metabolites in in vitro bioassays.

Correlation of sexual desire with sexual hormone binding globulin and free androgen index in women using combined contraceptives

Purpose To correlate the sexual desire levels with sexual hormone binding globulin and free androgen index in women taking different types of hormonal contraceptives (HCs) containing ethinylestradiol (EE), oestradiol valerate (E2V), 17β-oestradiol (E2), or estetrol (E4), combined or in phasic formulation with different progestogens having antiandrogenic properties. Methods Three hundred and sixty-seven women (age range 18-46) participated in the study. SHBG and total testosterone (TT) were measured, and the Free Androgen Index (FAI) was calculated. The Female Sexual Function Index (FSFI) and the Female Sexual Distress Scale (FSDS) questionnaires were used to assess sexual function and distress, respectively. Results The highest SHBG values and the lowest FAIs were obtained of women on HCs containing EE than those of women on HCs containing E2V/17β E2 or E4 (p < 0.001). Desire scores and FSFI total scores were lower in women on HCs with EE than in those using HCs containing E2V, 17β E2, or E4 (p ≤ 0.001). The women who were on HCs containing EE reported FSDS levels higher than those containing all the other types of oestrogen. Finally, sexual desire and FSFI total scores had a negative correlation with the SHBG values and a positive correlation with FAI percentage (p ≤ 0.0001). Conclusions A minority of women using HCs with EE might experience a decreased sexual desire. This was not observed in women on HCs containing E2V, 17 E2, or E4. To avoid HC discontinuation, due to sexual desire reduction, HCs having minor antiandrogenic effects could be taken into consideration.

Evaluation of Derma Genie™-H001 as a Functional Cosmetic Ingredient for Hair Loss Improvement: Assessing Anti-inflammatory, Antioxidative, Photoprotective, Antiandrogenic, and Hair Growth-Inductive Effects

Purpose: This study aimed to assess the potential of Derma Genie™-H001 as a functional cosmetic ingredient for alleviating hair loss symptoms by evaluating its impact on anti-inflammatory, antioxidative, photoprotective, antiandrogenicity, and activation of hair growth-related gene expression. Methods: We employed various methods, including nitric oxide (NO) assay and quantitative real-time polymerase chain reaction (qRT-PCR, to estimate anti-inflammatory activities). Additionally, this study assessed antioxidative and photoprotective effects on human dermal papilla cells (HDPCs) using water-soluble tetrazolium salt (WST-1) assay, 2′,7′-dichlorofluorescin diacetate (DCF-DA) assay, lactate dehydrogenase (LDH) assay, and ultraviolet B (UVB)-irradiation assay. To investigate the anti-hair loss effects of Derma Genie™-H001, 3D spheroid culture and qRT-PCR were conducted. Results: Derma Genie™-H001 exhibited the characteristics of polyphenols and demonstrated 2,2-dipheny-1-picrylhydrazyl (DPPH) radical scavenging activity. Importantly, it showed no cytotoxicity on HDPCs and increased the diameter of HDPCs 3D spheroid. Inflammatory mediators such as NO and proinflammatory cytokines were reduced in a dose-dependent manner in Raw 264.7 cells treated with Derma Genie™. Moreover, Derma Genie™ exhibited substantial protective effects against hydrogen peroxide (H&lt;sub&gt;2&lt;/sub&gt;O&lt;sub&gt;2&lt;/sub&gt;) and UVB-induced cytotoxicity in HDPCs. Notably, qRT-PCR analysis revealed a decrease in the expression of genes related to hair loss and an increase in the expression of genes associated with hair growth in HDPCs treated with Derma Genie™. Conclusion: Derma Genie™-H001 demonstrated promising effects in mitigating LPS-induced proinflammatory cytokines, H&lt;sub&gt;2&lt;/sub&gt;O&lt;sub&gt;2&lt;/sub&gt;-induced cytotoxicity, UVB-induced cytotoxicity, and DHT-induced TGF-β1 expression. Furthermore, it significantly upregulated the expression of hair growth-related genes. These findings suggest that Derma Genie™ may serve as a valuable functional cosmetic ingredient for improving hair loss symptoms.

Antiprogestins for breast cancer treatment: We are almost ready

The development of antiprogestins was initially a gynecological purpose. However, since mifepristone was developed, its application for breast cancer treatment was immediately proposed. Later, new compounds with lower antiglucocorticoid and antiandrogenic effects were developed to be applied to different pathologies, including breast cancer. We describe herein the studies performed in the breast cancer field with special focus on those reported in recent years, ranging from preclinical biological models to those carried out in patients. We highlight the potential use of antiprogestins in breast cancer prevention in women with BRCA1 mutations, and their use for breast cancer treatment, emphasizing the need to elucidate which patients will respond. In this sense, the PR isoform ratio has emerged as a possible tool to predict antiprogestin responsiveness. The effects of combined treatments of antiprogestins together with other drugs currently used in the clinic, such as tamoxifen, CDK4/CDK6 inhibitors or pembrolizumab in preclinical models is discussed since it is in this scenario that antiprogestins will be probably introduced. Finally, we explain how transcriptomic or proteomic studies, that were carried out in different luminal breast cancer models and in breast cancer samples that responded or were predicted to respond to the antiprogestin therapy, show a decrease in proliferative pathways. Deregulated pathways intrinsic of each model are discussed, as well as how these analyses may contribute to a better understanding of the mechanisms involved.

Cardamonin-loaded liposomal formulation for improving percutaneous penetration and follicular delivery for androgenetic alopecia.

Androgenic alopecia (AGA) has a considerable impact on the physical and mental health of patients. Nano preparations have apparent advantages and high feasibility in the treatment of AGA. Cardamonin (CAR) has a wide range of pharmacological activities, but it has the problems of poor solubility in water and low bioavailability. There are few, if any, researches on the use of nano-loaded CAR to improve topical skin delivery of AGA. In this study, a CAR-loaded liposomal formulation (CAR@Lip and CAR@Lip Gel) was developed and characterized. The prepared CAR@Lip exhibited a uniform and rounded vesicle in size. CAR@Lip and CAR@Lip Gel can significantly improve the cumulative release of CAR. Additionally, CAR@Lip can obviously promote the proliferation and migration of human dermal papilla cells (hDPCs). Cell uptake revealed that the uptake of CAR@Lip significantly increased compared with the free drug. Furthermore, both CAR@Lip and CAR@Lip Gel groups could markedly improve the transdermal performance of CAR, and increase the topical content of the drug in the hair follicle compared with CAR. The ratchet effect of hair follicles could improve the skin penetration depth of nanoformulations. Notably, Anti-AGA tests in the mice showed that CAR@Lip and CAR@Lip Gel groups could promote hair growth, and accelerate the transition of hair follicles to the growth stage. The anti-androgen effect was revealed by regulating the expression of IGF-1, VEGF, KGF, and TGF-β, participating in SHH/Gli and Wnt/β-catenin pathways. Importantly, the nanoformulations had no obvious skin irritation. Thus, our study showed that CAR-loaded liposomal formulation has potential application in the treatment of AGA.

The changing landscape of mineralocorticoid receptor antagonists – finerenone in cardiovascular and renal health

Finerenone is a novel non-steroidal mineralocorticoid receptor antagonist (MRA) with high selectivity and affinity to mineralocorticoid receptor (MR). Steroidal MRAs, like spironolactone and eplerenone, have been in use for decades. They have an established position in the management of hypertension and heart failure with reduced ejection fraction (HFrEF). There are studies showing that MR antagonism has anti-inflammatory and anti-fibrotic effects resulting in cardiovascular and renal protection. However, broader use of steroidal MRAs is seriously limited by antiandrogenic side effects and the risk of hyperkalemia. The differences in structure as well as pharmacokinetic and pharmacodynamic properties between steroidal and non-steroidal MRAs result in reduced risk of side effects while offering the benefit of cardiorenal protection. Finerenone is currently the only non-steroidal MRA approved by European Medicines Agency (EMA). It is indicated for patients with chronic kidney disease (CKD) with albuminuria and type 2 diabetes mellitus (DM2). Several studies showed a reduction of the risk of cardiovascular and renal events in this group of patients compared to placebo. The risk of hyperkalemia was increased resulting in discontinuation of treatment in more patients than placebo, but the risk of serious adverse events did not differ significantly. The other indications for finerenone, including heart failure (HF) management, are currently being researched.

The association between phthalate exposure and pubertal development.

Our results suggest that phthalate exposure, especially exposure to the HMWP, may have inverse association with male pubertal development. Further investigation is required to verify the relationship of phthalate exposure and pubertal development in girls. • Exposure to phthalates may have antiandrogenic effects. • Studies on the association between phthalates and pubertal development have yielded inconsistent results. • Phthalate levels were inversely associated with self-reported pubertal stages in boys. • Exposure to phthalates might have a negative influence on male pubertal development.

Gαi2 Protein Inhibition Blocks Chemotherapy- and Anti-Androgen-Induced Prostate Cancer Cell Migration.

We have previously shown that heterotrimeric G-protein subunit alphai2 (Gαi2) is essential for cell migration and invasion in prostate, ovarian and breast cancer cells, and novel small molecule inhibitors targeting Gαi2 block its effects on migratory and invasive behavior. In this study, we have identified potent, metabolically stable, second generation Gαi2 inhibitors which inhibit cell migration in prostate cancer cells. Recent studies have shown that chemotherapy can induce the cancer cells to migrate to distant sites to form metastases. In the present study, we determined the effects of taxanes (docetaxel), anti-androgens (enzalutamide and bicalutamide) and histone deacetylase (HDAC) inhibitors (SAHA and SBI-I-19) on cell migration in prostate cancer cells. All treatments induced cell migration, and simultaneous treatments with new Gαi2 inhibitors blocked their effects on cell migration. We concluded that a combination treatment of Gαi2 inhibitors and chemotherapy could blunt the capability of cancer cells to migrate and form metastases.

Evaluation of the anti-androgenic and cytotoxic effects of benzophenone-3 in male Sprague-Dawley rats

ABSTRACT Benzophenone-3 (BP-3, 2-hydroxy-4-methoxybenzophenone, oxybenzone) is one of the most widely used types of benzophenone organic sunscreen. However, this compound is a potentially harmful toxicant. The aim of this study was 2-fold to: (1) utilize a Hershberger bioassay in vivo in castrated male Sprague-Dawley rats to investigate the anti-androgenic activities of BP-3, and (2) use in vitro a methyl tetrazolium assay to compare the toxicity between Leydig cells (TM3 cells) and mouse fibroblast (NIH-3T3) cell lines. In the Hershberger assay, rats were divided into 6 groups (each of n = 7): a vehicle control, negative control, positive control, PB-3 low (40 mg/kg), BP-3 intermediate (200 mg/kg), and BP-3 high (1000 mg/kg)-dose. The weight of the ventral prostate was significantly decreased at BP-3 doses of 200 or 1,000 mg/kg/day. In addition, the levator anibulbocavernosus muscle weights were also significantly reduced at BP-3 doses of 40, 200, or 1,000 mg/kg/day. In the MTT assay, the viability of NIH-3T3 mouse fibroblast cells was within the normal range. However, the TM3 mouse testis Leydig cell viability was significantly lowered in a concentration-dependent manner. Therefore, data indicate that BP-3 might exert in vivo anti-androgenic and in vitro cytotoxic effects in cells associated with the male reproductive system compared to normal non-reproductive cells. Abbreviation: BP-3: benzophenone-3; CG: Cowper’s gland; DMEM: Dulbecco’s modified Eagle’s medium; DMSO: dimethyl sulfoxide; GP: glans penis; LABC: levator anibulbocavernosus muscle; MTT: methyl tetrazolium; NC: negative control; PC: positive control; SV: seminal vesicle; TP: testosterone propionate; VC: vehicle control; VP: ventral prostate

Ganoderma lucidum and Hypericum perforatum Exhibit Anti-Inflammatory, Antioxidant, and Anti-Androgen Effect in Rat Model of Experimental Polycystic Ovarian Syndrome

Objective: Chronic inflammation is considered to be of key importance in the pathogenesis of polycystic ovarian syndrome (PCOS). Ganoderma lucidum polysaccharide (GLP) and Hypericum perforatum (HP) have been reported to have anti-inflammatory and antioxidant activities. We studied the effects of these agents on ovarian tissue in a rat model of experimental PCOS. Materials and Methods: Forty-two Sprague-Dawley female rats were divided into 6 groups with 7 animals in each group as listed below: Group 1: Control, Group 2: PCOS, Group 3: PCOS + HP, Group 4: HP only, Group 5: PCOS+ GLP, Group 6: GLP only. At the end of the experimental procedures, all the animals underwent bilateral oophorectomy and blood samples were collected. Ovarian tissue and blood samples were used for biochemical and histopathological analysis. Results: Follicle degeneration in the PCOS group showed a statistically significant increase compared to the other groups (p < 0.05). Cystic follicles were significantly reduced in the PCOS+GLP and PCOS+HP groups as compared to the PCOS group. Levels of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) were elevated in PCOS rats (p < 0.01). Levels of sex hormone binding globulin (SHBG) levels were diminished (p < 0.01). Levels of malondialdehyde (MDA) and insulin-like growth factor 1 (IGF-1) were increased in PCOS rats as compared to the other groups (p < 0.02, p < 0.02, respectively). GLP supplementation diminished the levels of IGF-1 and MDA. GLP or HP supplementation increased reduced glutathione (GSH). Conclusion: GLP and HP treatment normalizes SHBG levels while correcting PCOS-induced hyperandrogenemia. Both herbs regulate the redox balance by decreasing the levels of MDA and increasing the level of GSH.

Современные возможности лечения доброкачественной гиперплазии предстательной железы и симптомов нижних мочевыводящих путей: новые данные и опыт клинического применения гексанового экстракта плодов Serenoa repens

The article discusses the pharmacological properties and results concerning the clinical use of a medicinal herbal preparation — hexane extract of Serenoa repens (HESr) (creeping palm fruit). In recent years, quite a lot of research data has been accumulated indicating a multicomponent mechanism of drug action. It has been found to have antiandrogenic, antiproliferative and anti-inflammatory effects in the tissues of the prostate gland (pancreas). Studies have been published describing a new mechanism of action of HESr — antineurogenic, the implementation of which helps to reduce infravesical obstruction, leads to inhibition of α1-adrenergic, thromboxane-induced and cholinergic contraction of the pancreatic and detrusor smooth muscles, and also inhibits the growth of pancreatic stromal cells. At present, there is evidence proving the highest clinical efficacy level of the HESr (data from several meta-analyses). The listed effects and patterns of HESr are illustrated by clinical examples of prescribing this drug to patients with lower urinary tract symptoms and benign pancreatic hyperplasia. The safety of use in conditions of polypragmasy in a comorbid patient, the possibility of long-term use without the adverse events occurrence and ensuring a higher life quality while maintaining full sexual function are also particularly noted. KEYWORDS: lower urinary tract symptoms, benign prostatic hyperplasia, chronic bacterial prostatitis, hexane extract of Serenoa repens fruit, ejaculation, life quality. FOR CITATION: Amdiy R.E., Simanov R.N. Modern treatment options for benign prostatic hyperplasia and lower urinary tract symptoms: new data and clinical picture of the hexane extract of Serenoa repens fruit. Russian Medical Inquiry. 2024;8(4):228–234 (in Russ.). DOI: 10.32364/2587-6821-2024-8-4-7.

Endocrine Disruptors and Estrogens in Human Prostatic Tissue.

Endocrine disruptors (EDs) are ubiquitous substances both in the environment and everyday products that interfere with the hormonal system. Growing evidence demonstrates their adverse effects on the organism, including the reproductive system and the prostate, owing to their (anti)estrogenic or antiandrogenic effects. Since EDs can interact with steroid hormone actions on-site, understanding the levels of intraprostatic EDs in conjunction with steroids may hold particular significance. The aim of this study was to develop and validate a method for determining estrogens, various groups of EDs (bisphenols, parabens, oxybenzone and nonylphenol) and phytoestrogens in their unconjugated and conjugated forms in prostate tissue by liquid chromatography-tandem mass spectrometry, and subsequently analyze 20 human prostate tissue samples. The method enabled 20 compounds to be analyzed: estrogens (estrone, estradiol, estriol), bisphenols (bisphenol A- BPA, BPS, BPF, BPAF, BPAP, BPZ, BPP), parabens (methyl-, ethyl-, propyl-, butyl-, benzyl- paraben), oxybenzone, nonylphenol and phytoestrogens (daidzein, genistein, equol) with LLOQs between 0.017-2.86 pg/mg of tissue. The most frequently detected EDs in prostate tissues were propylparaben (conjugated and unconjugated forms in 100 % of tissues), methylparaben (unconjugated in 45 % and conjugated in 100 %), ethylparaben (unconjugated in 25 % and conjugated in 100 % BPA (unconjugated in 35 % and conjugated in 60 % and oxybenzone (both forms in 45 % To the best of our knowledge, this is the first study detecting EDs, phytoestrogens and estriol conjugate (E3C) in the prostate. E3C was the most abundant estrogen in prostatic tissue. This highlights the need for further explorations into estrogen metabolism within the prostate.

Cyanidin-3-O-glucoside and its derivative vitisin A alleviate androgenetic alopecia by exerting anti-androgen effect and inhibiting dermal papilla cell apoptosis

Androgenetic alopecia (AGA), one of the most common forms of hair loss, lacks satisfactory treatment methods in modern society. This study employed an experimental design combining in vitro and in vivo approaches to explore the effects of Cyanidin-3-O-glucoside (C3G) and Carboxypyranocyanidin-3-O-glucoside (Vitisin A) on AGA. In human dermal papilla cells (HDPCs), both anthocyanins demonstrated inhibitory effects on androgen receptors, significantly reduced dihydrotestosterone (DHT) induced apoptosis of HDPCs, and regulated the secretion of Fibroblast growth factor 7 and Transforming growth factor beta 1. In vitro transdermal experiment revealed that both C3G and Vitisin A could penetrate mice skin, aided by the application of cream. Furthermore, in vivo experiments with mice indicated that application of C3G or Vitisin A cream effectively improved hair follicles miniaturization, regression, and apoptosis caused by DHT. The repression of Wnt10b and β-catenin expression induced by DHT was prevented by C3G and Vitisin A in both cell and mouse model. Consequently, these findings suggest that C3G and Vitisin A could be considered as alternative methods for alleviating AGA.

Hormonal and haematological biomarkers as indicators of stress induced by Diuron herbicide toxicity on Clarias gariepinus (Burchell, 1822) sub-adults

Diuron is a globally used herbicide for weed control but has anti-androgenic effects on androgens (testosterone and androstenedione), antagonist effects on thyroid hormone signaling, and haematological effects due to their biotransformation in fish. Endocrine-disrupting biomarkers such as thyroid hormones, sex hormones, and haematological indices of Clarias gariepinus sub-adults exposed to sub-lethal diuron concentrations were studied over a 28-day period. C. gariepinus (n = 200) sub-adults were exposed to sub-lethal concentrations (0.00, 0.09, 0.18, 0.26, and 0.35 mg/L) of diuron. Changes in the hormonal and haematological profiles of the exposed fish were concentration and exposure duration-dependent. The thyroxine (T4), tri-iodothyronine (T3), and 17β-estradiol (E2) profiles decreased with an increase in concentration and exposure duration. The haemoglobin, pack cell volume, red blood cell, white blood cell, mean cell volume, and mean corpuscular haemoglobin cell decreased, while the mean corpuscular haemoglobin increased with an increase in concentration and exposure duration. Diuron induced stress and altered the physiological mechanisms of fish, and its application in farmlands should be regulated so as to enable a sustainable aquatic eco-system and fishery resources.

In vitro antiandrogenic effects of the herbicide linuron and its metabolites

Although the herbicide linuron is banned for use in the EU due to its reproductive and developmental toxicity, it can still be found in randomly sampled foods grown in and outside the EU. It is not clear if metabolites of linuron can contribute to the endocrine disrupting effects following exposure to the parent compound. To address this gap, we analysed linuron and the metabolites 1-(3,4-dichlorophenyl) urea (DCU), 3,4-dichloroaniline (DCA) and 1-(3,4-dichlorophenyl)-3-methoxyurea (DCXU) for androgen receptor (AR) activities and effects on steroidogenesis. Generally, linuron and the metabolites showed qualitatively similar antiandrogenic profiles, but potencies varied. All compounds were AR antagonists, with linuron showing highest potency (IC50 of 2.8 μM). The overall picture of effects on steroidogenesis showed that linuron and metabolites increased the levels of estrogens and corticosteroids, whereas the synthesis of androgens was inhibited. The metabolite DCU was by far the most potent inhibitor of testosterone synthesis (IC50 of 6.7 μM compared to IC50 of 51.1 μM for linuron). We suggest that it is likely that the metabolites contribute to the antiandrogenic effects of linuron in vivo, especially by inhibiting testosterone synthesis.

Bisphenol A analogues and metabolic syndrome in women with polycystic ovary syndrome

Bisphenols (BPs) have become a chemical group of special interest due to their ability to interfere with the endocrine system and their ubiquitous presence in the environment. As some of them possess mild estrogenic and anti-androgenic effects, they might be associated with the diagnosis of polycystic ovary syndrome (PCOS). Acting on multiple tissues, BPs exposure may lead to metabolic derangements characteristic for metabolic syndrome (MetS). Therefore, the aim of this study was to determine the potential relationship between exposure to some BPA analogues and features of the MetS in women with PCOS. Serum BPE, BPC, BPG, BPM, BPP, BPZ, BPFL, and BPBP concentrations did not differ significantly between the PCOS (n = 135) and the control subjects (n = 104). However, women whose serum BPM and BPP concentrations were in the highest tertile were more likely to be diagnosed with PCOS (adjusted OR; [95%CI] 0.43; [0.20; 0.89], P < 0.001 and 0.56; [0.27; 0.96], P = 0.049, consequently). Serum concentrations of BPs were not associated with the MetS diagnosis in the PCOS group. There was a negative correlation between the concentrations of serum BPBP and total serum cholesterol (r = – 0.153; P = 0.019), BPE and serum testosterone (r = – 0.160; P = 0.014) as well as BPC and HDL-cholesterol (r = – 0.138; P = 0.036). There was a positive correlation between the concentrations of BPP and serum triglycerides (r = 0.138; P = 0.036). Our results point to the potential association between exposure to BPM, BPP, and the diagnosis of PCOS, along with the impact of BPBP, BPE, BPC, and BPP on the metabolic features of the MetS.

THU544 Prostatic Acid Phosphatase Is Negatively Regulated By Androgens And Persists In Castrate-resistant Metastatic Prostate Cancer

Abstract Disclosure: V. Rajagopalan: None. A. Kirschenbaum: None. S. Yao: None. A. Levine: None. Abstract: Prostate cancer (PCa) is the most common cancer and second leading cause of cancer death in American men. Most patients with metastatic PCa respond initially to androgen deprivation therapy (ADT) but the majority will progress to lethal, metastatic, castrate-resistant PCa (mCRPC). The identification of markers of mCRPC that are not regulated or negatively regulated by androgens can serve as markers and therapeutic targets in mCRPC. We hypothesized that prostatic acid phosphatase (PAP), a protein phosphatase and 5’ectonucleotidase that is expressed in both the cytoplasm and transmembrane (TM-PAP) of PCa cells, is negatively regulated by androgen signaling and can serve as a therapeutic target. Methods: Protein expression of PAP was assessed with western blotting in PCa cell lines. TM-PAP is identified by sub-cellular fractionation analysis, confocal microscopy, and flow cytometry. Effects of androgens and anti-androgens on PAP expression were assessed in VCaP cells with dihydrotestosterone (DHT) and Enzalutamide treatment followed by PAP, AR and PSA protein expression determined with Western Blotting. Loss of function analysis of PAP was performed with siRNA-mediated silencing and functional analysis was evaluated with migration and colony formation assays. Transcriptional regulation of PAP was determined by real time qPCR analysis. ChiPseq was done to confirm the binding of AR to PAP promoter. Results: All forms of PAP including TM-PAP are expressed in VCaP cell line. However, we did not detect PAP protein expression in-vitro in the LNCaP, C42B or 22Rv1 cell lines. In VCaP cell lines, we demonstrated that androgens decreased while anti-androgens increased PAP expression in a time and dose dependent fashion. Androgenic regulation of PAP occurred, at least in part, at the transcriptional level. We successfully knocked down TM and secretory PAP expression in VCaP cells using siRNA technology and showed that loss of PAP decreased migration and colony formation. Conclusions: The identification of markers of mCRPC that not regulated or negatively regulated by androgen could help identify recalcitrant tumor cells and mount targeted delivery of therapeutic agents. In the VCaP cell line, derived from a patient with mCRPC, TM and cytosolic PAP is one such protein that is negatively regulated by androgen signaling and may therefore serve as a viable therapeutic target in mCRPC. Presentation: Thursday, June 15, 2023

Reproductive toxicity following in utero and lactational exposure to a human-relevant phthalate mixture in rats.

This rodent (Wistar rats) study examined reproductive effects of in utero/lactational exposure to a mixture of 6 antiandrogenic phthalates (PMix): diisobutyl phthalate, di-n-butyl phthalate, diisopentyl phthalate, butylbenzyl phthalate, di-2-ethylhexyl phthalate, and diisononyl phthalate. The PMix was defined based on exposure data from pregnant women in Brazil. Experimental groups were established by extrapolating the estimated human dose to rats (0.1 mg/kg/day), followed by up to 3 additional doses corresponding to 5, 1000, and 5000 times the starting rat dose: 0 (control), 0.1, 0.5, 100, and 500 mg/kg/day. The fetal experiment assessed gestational exposure effects on fetal gonads, whereas the postnatal experiment evaluated reproductive parameters in males and females after in utero and lactational exposure. Prenatal exposure decreased fetal testicular testosterone production at 0.5 and 500 mg/kg/day. PMix 500 also reduced mRNA expression of steroidogenesis-related genes, upregulated transcript expression of the retinoic acid-degrading enzyme Cyp26b1, and increased multinucleated gonocytes incidence in fetal testes. Postnatal assessment revealed antiandrogenic effects at the highest dose, including reduced anogenital distance, nipple retention, and decreased weight of reproductive organs. Early puberty onset (preputial separation) was observed at the lowest dose in males. In contrast, females did not show significant changes in fetal and adult endpoints. Overall, the PMix recapitulated early and late male rat phthalate syndrome phenotypes at the highest dose, but also induced some subtle changes at lower doses, which warrant confirmation and mechanistic assessments. Our data support the use of epidemiologically defined mixtures for exposure risk assessments over traditional toxicological approaches.

Single and mixture toxicity evaluation of avobenzone and homosalate to male zebrafish and H295R cells

Avobenzone and homosalate are widely used in sunscreens to provide ultraviolet (UV) protection, either as single compounds or in combination. Some UV filters exhibit estrogenic or anti-androgenic activities, however, studies regarding their interactions and toxicity in mixtures are limited. In this study, the effect of the toxicity of a binary mixture comprising avobenzone (0.72 μg L−1) and homosalate (1.02 and 103 μg L−1) on steroid hormone biosynthesis were investigated using male zebrafish and human adrenocortical carcinoma (H295R) cells. In fish exposed to homosalate, a significant decrease in the gonadosomatic index, testosterone level, and transcription of several genes (e.g, hsd3b2, cyp17a1, and hsd17b1) and a significant increase in the hepatosomatic index, liver steatosis, 17β-estradiol level, and transcription of vtg gene were observed. These results suggest that estrogenic and anti-androgenic effects of homosalate were mediated by the steroidogenic pathway. The presence of 0.72 μg L−1 of avobenzone augmented the anti-androgenic responses in male fish. The testosterone level in the H295R cells were significantly decreased after they were exposed to homosalate alone or in combination with avobenzone, which is consistent with observations in male zebrafish. Further studies need to be conducted to understand the endocrine disrupting properties of long-term exposure to substances typically used in sunscreens.

The benefits of estetrol addition to drospirenone for contraception

Ethinylestradiol and drospirenone combined oral contraceptive formulations have been marketed for >20 years. Drospirenone has antimineralocorticoid and anti-androgenic effects that may offer several health benefits. Recently, 2 new drospirenone-containing oral contraceptives entered the market, 1 as a progestin-only pill containing 4 mg drospirenone and the other as a combined oral contraceptive containing 15 mg estetrol and 3 mg drospirenone. Estetrol has a unique differential effect on nuclear and membrane estrogen α-receptors when compared with other estrogens, leading to low impact on the liver, breast, and hemostasis parameters and a beneficial effect on the endometrium, vagina, cardiovascular system, bone, and brain. Phase 3 clinical studies demonstrated that the Pearl Index (pregnancies per 100-woman-years) for drospirenone alone is 4.0 in the United States and 0.93 in the European Union and for the estetrol-drospirenone combination it is 2.65 and 0.44, respectively. Drospirenone alone demonstrates high rates of unscheduled bleeding and low rates of scheduled bleeding, whereas the estetrol-drospirenone combination demonstrates a predictable and regular bleeding profile for most users with a high stable rate of scheduled bleeding and a low rate of unscheduled bleeding, reported primarily as spotting only. The adverse event profiles and discontinuation rates owing to adverse events are comparable, and no clinically significant effects were observed on metabolic parameters with either product. Hemostatic assays for drospirenone do not fully evaluate all parameters although the testing that is available suggests negligible effects, whereas validated hemostatic assays demonstrate that the estetrol-drospirenone combination has limited impact on hemostasis. The introduction of 4 mg drospirenone and 15 mg estetrol with 3 mg drospirenone are valuable additions to the contraceptive market. Adding estetrol to 3 mg drospirenone provides advantages of contraceptive efficacy and a regular, predictable bleeding profile with minimal impact on hemostasis parameters.