Androgen Production Research Articles

PACS-1 Interacts with TRPC3 and ESyt1 to Mediate Protein Trafficking while Promoting SOCE and Cooperatively Regulating Hormone Secretion.

Corticotropic cells of the anterior pituitary gland release adrenocorticotropic hormone (ACTH) in a regulated manner to promote the production of cortisol and androgens. The process of ACTH secretion is partly mediated by the phosphofurin acidic cluster sorting protein 1 (PACS-1); however, the underlying mechanisms behind this regulation remain unclear. Herein, we demonstrated PACS-1 interactions with the short transient receptor potential channel 3 (TRPC3) calcium transporter and the extended synaptotagmin-1 (ESyt1) endoplasmic reticulum-plasma membrane tethering protein. Importantly, PACS-1 promoted interactions between TRPC3 and ESyt1 and regulated their plasma membrane localization. Lastly, we demonstrated that PACS-1 is required for a proper store-operated calcium entry (SOCE) response and that ESyt1 regulates ACTH secretion through an unknown mechanism regulated by PACS-1. Overall, our study provides new insights into the physiological role PACS-1 plays in modulating intracellular calcium levels and regulating ACTH secretion in corticotropic cells.

The role of asprosin in regulating ovarian granulosa- and theca-cell steroidogenesis: a review with comparisons to other adipokines.

Adipose tissues produce a variety of biologically active compounds, including cytokines, growth factors and adipokines. Adipokines are important as they function as endocrine hormones that are related to various metabolic and reproductive diseases. The goal of this review was to summarise the role of asprosin, a recently discovered adipokine, and compare its role in ovarian steroidogenesis with that of other adipokines including adiponectin, leptin, resistin, apelin, visfatin, chemerin, irisin, and gremlin 1. The summary of concentrations of these adipokines in humans, rats and other animals will help researchers identify appropriate doses to test in future studies. Review of the literature indicated that asprosin increases androstenedione production in theca cells (Tc), and when cotreated with FSH increases oestradiol production in granulosa cells (Gc). In comparison, other adipokines (1) stimulate Gc oestradiol production but inhibit Tc androgen production (adiponectin), (2) inhibit Gc oestradiol production and Tc androstenedione production (leptin and chemerin), (3) inhibit Gc steroidogenesis with no effect on Tc (resistin), (4) inhibit Gc oestradiol production but stimulate Tc androgen production (gremlin 1), and (5) increase steroid secretion by Gc, with unknown effects on Tc steroidogenesis (apelin and visfatin). Irisin has direct effects on Gc but its precise role (inhibitory or stimulatory) may be species dependent and its effects on Tc will require additional research. Thus, most adipokines have direct effects (either positive or negative) on steroid production in ovarian cells, but how they all work together to create a cumulative effect or disease will require further research.

Exploring the Contribution of Traditional Medicinal Herbs in ManagingPolycystic Ovary Syndrome: A Comprehensive Review

: Polycystic ovary syndrome (PCOS) is a significant gynecological disorder that commonly affects women of reproductive age. Disturbances in the production and metabolism of androgens and estrogens mark the condition. Due to the rising occurrence of PCOS and its related physical and psychological issues, along with the role of sex hormone fluctuations in its development, the use of marketed drugs is effective in treating PCOS but is accompanied by negative consequences. Consequently, a significant number of individuals with PCOS are opting for natural remedies due to the positive therapeutic effects associated with natural medications and the constraints of allopathic treatments. This review aims to assess the impact of different herbs on alterations in sex hormone levels in the bloodstream and ovarian tissue. By conducting a comprehensive literature search utilizing databases such as PubMed, Google Scholar, and Crossref, we have gathered and analyzed pertinent information regarding the efficacy of herbal remedies in combating PCOS. Based on the available evidence, herbs consisting of phytoestrogens can potentially reduce insulin resistance, hyperandrogenism, and ovarian weight while facilitating ovulation. This review explores the importance of herbal remedies in addressing PCOS, elucidates their mechanisms of action, and examines the therapeutic uses of specific herbal medications to treat PCOS. Human studies on medicinal herbs for managing Polycystic Ovary Syndrome (PCOS) need extended durations for a comprehensive evaluation of both safety and efficacy, emphasizing the necessity for more long-term research. It is anticipated that this comprehensive review will serve as a valuable resource for researchers studying the effect of herbs on PCOS, providing them with valuable insights and information.

In Vitro Generation of Haploid Germ Cells from Human XY and XXY Immature Testes in a 3D Organoid System.

Increasing survival rates of children following cancer treatment have resulted in a significant population of adult survivors with the common side effect of infertility. Additionally, the availability of genetic testing has identified Klinefelter syndrome (classic 47,XXY) as the cause of future male infertility for a significant number of prepubertal patients. This study explores new spermatogonia stem cell (SSC)-based fertility therapies to meet the needs of these patients. Testicular cells were isolated from cryopreserved human testes tissue stored from XY and XXY prepubertal patients and propagated in a two-dimensional culture. Cells were then incorporated into a 3D human testicular organoid (HTO) system. During a 3-week culture period, HTOs maintained their structure, viability, and metabolic activity. Cell-specific PCR and flow cytometry markers identified undifferentiated spermatogonia, Sertoli, Leydig, and peritubular cells within the HTOs. Testosterone was produced by the HTOs both with and without hCG stimulation. Upregulation of postmeiotic germ cell markers was detected after 23 days in culture. Fluorescence in situ hybridization (FISH) of chromosomes X, Y, and 18 identified haploid cells in the in vitro differentiated HTOs. Thus, 3D HTOs were successfully generated from isolated immature human testicular cells from both euploid (XY) and Klinefelter (XXY) patients, supporting androgen production and germ cell differentiation in vitro.

Protective effect of Chlorella vulgaris on testicular damage, sperm parameters, androgen production, apoptosis and oxidative stress index in male rats following doxorubicin administration

Doxorubicin (DOX) is a chemotherapy agent associated with adverse effects on male reproductive health. Chlorella vulgaris (ChV) is a potent natural antioxidant with promising applications in maintaining health and preventing oxidative stress-related diseases. The present study aimed to investigate the protective effect of ChV on DOX-induced testicular toxicity. Twenty-five Wistar rats (230 ± 20 g) were randomly assigned to five groups (n = 5), including the control group, sham group (received normal saline by oral gavage daily and intraperitoneally (IP) once a week), DOX group (3 mg/kg; once a week; IP), ChV group (300 mg/kg/day; by oral gavage), and DOX (3 mg/kg; once a week; IP) + ChV (300 mg/kg/day; by oral gavage) group. After 8 weeks of treatment, the rats were euthanized and serum testosterone level, testes histomorphometry, gonadosomatic index (GSI), apoptotic gene expression, oxidative stress index, and sperm parameters were assessed. The results showed that DOX led to a significant decrease in histological indexes, testosterone level, GSI, sperm parameters, and Bcl-2 gene expression and increased expression of P-53 and Bax genes, and oxidative stress markers (P<0.05). The administration of ChV in the DOX+ChV group significantly improved testosterone levels, sperm parameters, testicular tissue apoptosis, antioxidant enzymes, and structural integrity of the testes (P<0.05). The findings suggest that the co-administration of ChV can be a promising therapeutic agent to reduce the adverse effects of DOX on male reproductive performance.

Endocrine disruption and male reproductive disorders: unanswered questions.

Maternal exposure to endocrine-disrupting chemicals (EDCs) in human pregnancy is widely considered as an important cause of adverse changes in male reproductive health due to impaired foetal androgen production/action. However, the epidemiological evidence supporting this view is equivocal, except for certain phthalates, notably diethyl hexyl phthalate (DEHP). Maternal phthalate exposure levels associated with adverse reproductive changes in epidemiological studies are several thousand-fold lower than those needed to suppress foetal androgen production in rats, and direct studies using human foetal testis tissue show no effect of high phthalate exposure on androgen production. This conundrum is unexplained and raises fundamental questions. Human DEHP exposure is predominantly via food with highest exposure associated with consumption of a Western style (unhealthy) diet. This diet is also associated with increased exposure to the most common EDCs, whether persistent (chlorinated or fluorinated chemicals) or non-persistent (phthalates, bisphenols) compounds, which are found at highest levels in fatty and processed foods. Consequently, epidemiological studies associating EDC exposure and male reproductive health disorders are confounded by potential dietary effects, and vice versa. A Western diet/lifestyle in young adulthood is also associated with low sperm counts. Disentangling EDC and dietary effects in epidemiological studies is challenging. In pregnancy, a Western diet, EDC exposure, and maternal living in proximity to industrial sites are all associated with impaired foetal growth/development due to placental dysfunction, which predisposes to congenital male reproductive disorders (cryptorchidism, hypospadias). While the latter are considered to reflect impaired foetal androgen production, effects resulting from foetal growth impairment (FGI) are likely indirect. As FGI has numerous life-long health consequences, and is affected by maternal lifestyle, research into the origins of male reproductive disorders should take more account of this. Additionally, potential effects on foetal growth/foetal testis from the increasing use of medications in pregnancy deserves more research attention.

Assessment of prognostic factors in pediatric adrenocortical tumors: the modified pediatric S-GRAS score in an international multicenter cohort-a work from the ENSAT-PACT working group.

Pediatric adrenocortical carcinoma (pACC) is rare, and prognostic stratification remains challenging. We aimed to confirm the prognostic value of the previously published pediatric scoring system (pS-GRAS) in an international multicenter cohort. Analysis of pS-GRAS items of pACC from 6 countries in collaboration of ENSAT-PACT, GPOH-MET, and IC-PACT. We received patient data of the pS-GRAS items including survival information from 9 centers. PS-GRAS score was calculated as a sum of tumor stage (1 = 0; 2-3 = 1; 4 = 2 points), grade (Ki67 index: 0%-9% = 0; 10%-19% = 1; ≥20% = 2 points), resection status (R0 = 0; RX/R1/R2 = 1 point), age (<4 years = 0; ≥4 years = 1 point), and hormone production (androgen production = 0; glucocorticoid-/mixed-/no-hormone production = 1 point) generating 8 scores and 4 groups (1: 0-2, 2: 3-4, 3: 5, 4: 6-7). Primary endpoint was overall survival (OS). We included 268 patients with median age of 4 years. The analysis of the pS-GRAS score showed a significantly favorable prognosis in patients with a lower scoring compared to higher scoring groups (5-year OS: Group 1 98%; group 2 87% [hazard ratio {HR} of death 3.6, 95% CI of HR 1.6-8.2]; group 3 43% [HR of death 2.8, 95% CI 1.9-4.4]; group 4: OS 18% [HR of death 2.1, 95% CI 1.7-2.7]). In the multivariable analysis, age (HR of death 3.5, 95% CI 1.8-7.0), resection status (HR of death 5.5, 95% CI 2.7-11.1), tumor stage (HR of death 1.9, 95% CI of HR 1.2-3.0), and Ki67 index (HR of death 1.7, 95% CI 1.2-2.4) remained strong independent outcome predictors. Especially infants < 4 years showed more often low-risk constellations with a better OS for all tumor stages. In an international multicenter study, we confirmed that the pS-GRAS score is strongly associated with overall survival among patients with pACC. Age, resection status, stage, and Ki67 index are important parameters for risk stratification.

Mitochondrial dysfunction results in enhanced adrenal androgen production in H295R cells

The role of mitochondria in steroidogenesis is well established. However, the specific effects of mitochondrial dysfunction on androgen synthesis are not fully understood. In this study, we investigate the effects of various mitochondrial and metabolic inhibitors in H295R adrenal cells and perform a comprehensive analysis of steroid and metabolite profiling. We report that mitochondrial complex I inhibition by rotenone shifts cells toward anaerobic metabolism with a concomitant hyperandrogenic phenotype characterized by rapid stimulation of dehydroepiandrosterone (DHEA, 2 h) and slower accumulation of androstenedione and testosterone (24 h). Screening of metabolic inhibitors confirmed DHEA stimulation, which included mitochondrial complex III and mitochondrial pyruvate carrier inhibition. Metabolomic studies revealed truncated tricarboxylic acid cycle with an inverse correlation between citric acid and DHEA production as a common metabolic marker of hyperandrogenic inhibitors. The current study sheds light on a direct interplay between energy metabolism and androgen biosynthesis that could be further explored to identify novel molecular targets for efficient treatment of androgen excess disorders.

Prevention of Male Late-Onset Hypogonadism by Natural Polyphenolic Antioxidants.

Androgen production primarily occurs in Leydig cells located in the interstitial compartment of the testis. In aging males, testosterone is crucial for maintaining muscle mass and strength, bone density, sexual function, metabolic health, energy levels, cognitive function, as well as overall well-being. As men age, testosterone production by Leydig cells of the testes begins to decline at a rate of approximately 1% per year starting from their 30s. This review highlights recent findings concerning the use of natural polyphenolics compounds, such as flavonoids, resveratrol, and phenolic acids, to enhance testosterone production, thereby preventing age-related degenerative conditions associated with testosterone insufficiency. Interestingly, most of the natural polyphenolic antioxidants having beneficial effects on testosterone production tend to enhance the expression of the steroidogenic acute regulatory protein (Star) gene in Leydig cells. The STAR protein facilitates the entry of the steroid precursor cholesterol inside mitochondria, a rate-limiting step for androgen biosynthesis. Natural polyphenolic compounds can also improve the activities of steroidogenic enzymes, hypothalamus-pituitary gland axis signaling, and testosterone bioavailability. Thus, many polyphenolic compounds such as luteolin, quercetin, resveratrol, ferulic acid phenethyl ester or gigantol may be promising in delaying the initiation of late-onset hypogonadism accompanying aging in males.

Lifestyle changes and Nutrition in Polycystic Ovarian Disorder: A Holistic Review

Polycystic Ovarian Disorder (PCOD) is a prevalent endocrine illness in women of reproductive age which is a reproductive and metabolic disorder. It shows hormonal abnormalities in which there is an excessive production of androgens and that is one of the main causes of infertility, diabetes, irregular menstrual periods, hirsutism, hair loss, obesity and acne. One of the main features of PCOD is the presence of cysts on the ovaries, which can affect their ability to release eggs regularly and Anti-Müllerian hormone (AMH) plays an inhibitory role in follicular development and recruitment, contributing to follicular arrest. AMH inhibitory action on FSH-induced aromatase production likely contributes to hyperandrogenism in PCOD. Some factors which can cause of PCOD such as lack of exercise, skipping breakfast, fast food, unbalanced sleep, obesity, genetic, environmental factors and stress level. Lifestyle and nutrition can affect PCOD development and maintenance in PCOD can be done by lifestyle changes such as diet regulation (in taking of vitamins, antioxidant and Creatine) and exercise, as well as medications to regulate hormones, improve fertility and sleep which play a vital role in PCOD management.

How Per- and Poly-Fluoroalkyl Substances Affect Gamete Viability and Fertilization Capability: Insights from the Literature.

There has been emerging research linking per- and poly-fluoroalkyl substances (PFAS) to gamete viability and fertility. PFAS, prevalent in the environment and water supplies, undergo slow degradation due to their C-F bond and a long half-life (2.3-8.5 years). In females, PFAS inhibit the hypothalamic-pituitary-gonadal (HPG) axis, reducing follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels, leading to the inhibition of androgen and estradiol production. PFAS have been found to cause detrimental effects on egg quality through impairing folliculogenesis. In males, PFAS can impair sperm motility and morphology: two fundamental qualities of successful fertilization. PFAS exposure has been proven to inhibit testosterone production, sperm capacitation, and acrosomal reaction. After fertilization, the results of PFAS exposure to embryos have also been investigated, showing reduced development to the blastocyst stage. The aim of this review is to report the main findings in the literature on the impact of PFAS exposure to gamete competency and fertilization capability by highlighting key studies on both male and female fertility. We report that there is significant evidence demonstrating the negative impacts on fertility after PFAS exposure. At high doses, these environmentally abundant and widespread compounds can significantly affect human fertility.

A Dose-Response Study on Functional and Transcriptomic Effects of FSH on Ex Vivo Mouse Folliculogenesis.

Follicle-stimulating hormone (FSH) binds to its membrane receptor (FSHR) in granulosa cells to activate various signal transduction pathways and drive the gonadotropin-dependent phase of folliculogenesis. Both FSH insufficiency (due to genetic or nongenetic factors) and FSH excess (as encountered with ovarian stimulation in assisted reproductive technology [ART]) can cause poor female reproductive outcomes, but the underlying molecular mechanisms remain elusive. Herein, we conducted single-follicle and single-oocyte RNA sequencing analysis along with other approaches in an ex vivo mouse folliculogenesis and oogenesis system to investigate the effects of different concentrations of FSH on key follicular events. Our study revealed that a minimum FSH threshold is required for follicle maturation into the high estradiol-secreting preovulatory stage, and such threshold is moderately variable among individual follicles between 5 and 10 mIU/mL. FSH at 5, 10, 20, and 30 mIU/mL induced distinct expression patterns of follicle maturation-related genes, follicular transcriptomics, and follicular cAMP levels. RNA sequencing analysis identified FSH-stimulated activation of G proteins and downstream canonical and novel signaling pathways that may critically regulate follicle maturation, including the cAMP/PKA/CREB, PI3K/AKT/FOXO1, and glycolysis pathways. High FSH at 20 and 30 mIU/mL resulted in noncanonical FSH responses, including premature luteinization, high production of androgen and proinflammatory factors, and reduced expression of energy metabolism-related genes in oocytes. Together, this study improves our understanding of gonadotropin-dependent folliculogenesis and provides crucial insights into how high doses of FSH used in ART may impact follicular health, oocyte quality, pregnancy outcome, and systemic health.

Characterization of ovarian progenitor cells for their potential to generate steroidogenic theca cells in vitro.

Progenitor cells with ovulation-related tissue repair activity were identified with defined markers (LGR5, EPCR, LY6A, and PDGFRA), but their potentials to form steroidogenic cells were not known. This study shows that the cells can generate progenies with different steroidogenic activities. Adult mammalian ovaries contain stem/progenitor cells necessary for folliculogenesis and ovulation-related tissue rupture repair. Theca cells are recruited and developed from progenitors during the folliculogenesis. Theca cell progenitors were not well defined. The aim of current study is to compare the potentials of four ovarian progenitors with defined markers (LY6A, EPCR, LGR5, and PDGFRA) to form steroidogenic theca cells in vitro. The location of the progenitors with defined makers was determined by immunohistochemistry and immunofluorescence staining of ovarian sections of adult mice. Different progenitor populations were purified by magnetic-activated cell sorting (MACS) and/or fluorescence-activated cell sorting (FACS) techniques from ovarian cell preparation and were tested for their abilities to generate steroidogenic theca cells in vitro. The cells were differentiated with a medium containing LH, ITS, and DHH agonist for 12 days. The results showed that EPCR+ and LGR5+ cells primarily distributed along the ovarian surface epithelium (OSE), while LY6A+ cells distributed in both the OSE and parenchyma. However, PDGFRA+ cells were exclusively located in interstitial compartment. When the progenitors were purified by these markers and differentiated in vitro, LY6A+ and PDGFRA+ cells formed steroidogenic cells expressing both CYP11A1 and CYP17A1 and primarily producing androgens, showing characteristics of theca-like cells, while LGR5+ cells generated steroidogenic cells devoid of CYP17A1 expression and androgen production, showing a characteristic of progesterone-producing cells (granulosa- or lutea-like cells). In conclusion, progenitors from both OSE and parenchyma of adult mice are capable of generating steroidogenic cells with different steroidogenic capacities, showing a possible lineage preference.

Identification of hCG- and LH-dependent ovarian luteoma resulting in marked androgen production during pregnancy states as well as after menopause: characterization in vivo and in a human ex vivo model

Identification of hCG- and LH-dependent ovarian luteoma resulting in marked androgen production during pregnancy states as well as after menopause: characterization <i>in vivo</i> and in a human ex vivo model

High Salt Diet Regulates Intrarenal Sex Steroid Biosynthesis Pathway in a Sex-Specific Manner

Hypertension is more prevalent in men than age-matched pre-menopausal women. Sodium (Na+) homeostasis plays a major role in the control of blood pressure. Renal medullary estrogenic signaling promotes Na+ excretion in female, but not male Sprague Dawley (SD) rats implicating estrogen as a potential female-specific natriuretic factor. Recently we showed that the components of estrogen biosynthesis pathway are expressed in rat kidney suggesting the kidneys can produce estrogen locally to invoke this natriuretic response. It is unknown how dietary Na+ impacts biosynthesis of sex hormones. We hypothesize that a high salt (HS) diet enhances intrarenal estrogen biosynthesis pathway, and this enhancement is more pronounced in females compared to males. To test this hypothesis, we harvested the renal cortex from 14 week old male and female SD rats maintained on either a normal salt (NS 0.3% NaCl) or HS (4% NaCl) diet for 5 days. Given that cholesterol is the main precursor of steroid hormones, we measured the mRNA expression of HMG-CoA reductase (HMGCR), the key enzyme for cholesterol synthesis; and the steroidogenic acute regulatory protein (StAR), which transports cholesterol into the mitochondria for steroid commitment. We also assessed the mRNA expression of steroid 5α reductase (SRD5A), which converts testosterone to the non-aromatizable androgen, dihydrotestosterone. Further, we assessed aromatase, the key enzyme in estrogen biosynthesis, via immunohistochemistry. We found that renal HMGCR was more abundant in female compared to male rats maintained on a NS diet (P=0.0467). Increasing dietary salt intake upregulated renal HMGCR expression in male (P=0.0020), but not female rats (NS male; 100.0 ± 26.8 vs. NS female; 235.3 ± 23.2 vs. HS male 294.6 ± 42.0 vs HS female 314.6 ± 21.0 relative expression % NS male n=6-7/group). On a NS diet, renal StAR was more abundant in female compared to male rats (NS male; 100.0 ± 25.3 vs. NS female; 217.2 ± 36.5 relative expression % NS male P=0.0255 n=5-7/group). Increasing dietary salt did not alter expression of StAR in either sex. Similarly, renal SRD5A expression was higher in female compared to male (P=0.0141) rats. In addition, a HS diet increased the expression of SRD5A in female rats (0.0093) and tended to increase SRD5A expression in male (P=0.0849) rats (NS male; 100.0 ± 11.6 vs. NS female; 297.8 ± 37.7 vs. HS male 243.0 ± 39.2 vs HS female 498.8 ± 44.8 relative expression % NS male n=6-7/group). We also revealed that aromatase was expressed at a comparable level in male and female renal cortex, regardless of the diet (NS male; 2.4 ± 0.3 vs. NS female; 2.6 ± 0.3 vs. HS male 2.8 ± 0.3 vs HS female 1.9 ± 0.1 Average intensity x area mM2 n=6-9/group). Together, this data suggests that (i) the female kidney may be more primed for steroid biosynthesis and (ii) there is a sex-specific upregulation of the intrarenal sex steroid biosynthesis pathway in response to increasing dietary salt intake. Additional work is required to investigate the potential role of the kidney in androgen and estrogen production. A better understanding of this steroidogenic pathway could reveal novel sex-specific therapeutics for hypertension. T32DK007569-34 Carl Gottschalk Grant. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Unlocking Fertility - with administration of Lekhaniya Mahakashaya in Pushpaghani Jataharini w.s.r. to PCOD

Children are a gift from the Lord; the fruit of the womb is a divine reward. Average Reproductive Age is 14-49 yrs. This age group have number of conditions related to menstrual cycle, such as Menorrhagia, Dysmenorrhea, Irregular menses, PCOD, endometriosis, etc. According to WHO - PCOD is a lifestyle disorder which is a common hormonal condition that affects women of reproductive age. It usually starts during adolescence, but symptoms may fluctuate over time. PCOD can cause hormonal imbalances, irregular periods, excess androgen levels and cysts in the ovaries, absence of menstruation, excessive hair growth (face, body, including back, belly and chest), acne, weight gain, hair loss, skin darkening (in neck and groin). Irregular periods, usually with a lack of ovulation, can lead infertility. PCOD is a leading cause of infertility. PCOD can lead to anovulation where ovaries stop releasing eggs. PCOD is a common condition that causes 70% of anovulation cases. PCOD causes the excessive production of androgens, which cause the follicles in your ovaries to remain small instead of maturing and growing as they have to prepare for ovulation. In Ayurveda on the basis of symptoms Pushpaghani Jataharini can be correlated with PCOD. PCOD is a chronic condition and cannot be cured. However, some symptoms can be improved through lifestyle changes and medications. In order to treat PCOD the changes in the life style and the administrations of Ausadhies like Lekhaniya Mahakashaya (Musta, Kustha, Haridra, Daruharidra, Vacha, Ativisha, Katurohini, Chitrak, Chirbilva).

Abstract SY38-01: Diverse effects of obesity on cancer immunology and therapy: Both the good and the bad

Abstract Obesity (BMI of 30 or higher) has reached pandemic proportions in the US and continues to rise. Obesity is associated with increased cancer incidence, progression, and mortality. Obesity is also associated with a meta-inflammation as well as general immune suppression. However, we and others have observed in both preclinical models involving diet-induced obese (DIO) mice as well as clinical cancer outcomes, that obesity can be associated with increased efficacy and T cell responses following immune checkpoint inhibition (ICI) targeting PD-1/PDL-1 (Wang Z et al, Nat Med 2019). This protective effect in preclinical models occurred despite marked increased tumor growth and progression in DIO mice. The results indicated that an “obesity paradox” existed regarding some cancer immune therapies. However, this increased efficacy did not apply to all immune therapy responses as we have also observed that following strong systemic immunostimulatory regimens such as with high dose cytokines (IL2) or following an allogeneic hematopoietic stem cell transplant (HSCT), that obesity was correlated with poorer outcomes due to increased cytokine release syndrome (CRS) due to macrophage dysregulation, reduction in gut microbiome diversity, and increased gut permeability, in both preclinical models and clinical outcomes (Khuat LT et al, Sci Trans Med 2020). These results indicated that a balance exists between the positive and negative effects of obesity which could be contingent on the particular type of cancer immunotherapy applied. With ICI, obesity is beneficial, but strong systemic immunostimulatory therapies can result in heightened proinflammatory responses and pathologies. It has also been reported that sex differences regarding the obesity paradox may also occur with high BMI males demonstrating increased anti-tumor efficacy (McQuade JL et al, Lancet Oncol 2018). The physiology of obesity regarding adiposity deposition and metabolic perturbations are also affected by sex hormones and obesity can also have marked effects on both androgen and estrogen production. We therefore assessed the role of sex on different cancer immunotherapy outcomes in obesity. Following systemic cytokine application or after allogeneic HSCT, no differences were observed between male and female DIO mice regarding the increased toxicities due to CRS. In marked contrast, following ICI targeting PD-1, increased anti-tumor effects were only observed in male and not female DIO mice. This was associated with immune differences in which female DIO mice had higher myeloid derived suppressor cells (MDSCs) and decreased T cell responses. However, when female mice were ovariectomized before placed on diet, application of ICI now resulted in increased anti-tumor efficacy comparable to DIO males. Importantly, metabolomics indicated that key metabolite changes could be correlated with the return of responsiveness. Similarly, clinical outcomes stratified on both sex and BMI also demonstrated the increased efficacy following ICI in high BMI males. These results demonstrate that sex-linked differences exist regarding the effects obesity has on cancer immunotherapy outcome following ICI but not other types of immune therapies. Further, these sex-based differences in obesity can be correlated with immunological alterations, and these can be reversed by estrogen inhibition suggesting that hormone modulation may offer immunological benefits in the context of obesity following cancer immunotherapy involving ICI. Citation Format: William J. Murphy, Logan Vick, Robert Canter, Spencer Rosario, Arta Monjazeb. Diverse effects of obesity on cancer immunology and therapy: Both the good and the bad [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr SY38-01.

Androgenic Properties and Subchronic Toxicity of the Aqueous Extract of Pycnanthus angolensis (Welw.) Warb. Wood (Myristicaceae)

Aims: Pycnanthus angolensis (Welw.) Warb. also known as “Etengué” in the Baka language, is a medicinal plant used by the Baka Pygmies of Cameroon to treat erectile dysfunction. This study aimed to evaluate the androgenic activity and subchronic toxicity of the aqueous extract of Pycnanthus angolensis wood.&#x0D; Study Design: Experimental design.&#x0D; Place and Duration of Study: Laboratory of Biochemistry, Faculty of Medicine and Biomedical Sciences, University of Yaoundé I, between November 2022 and October 2023.&#x0D; Methodology: Twenty-five male Wistar strain rats were divided into 5 groups of 5 rats each including a positive control group that received testosterone enanthate (5 mg/kg) intramuscularly once a week; a normal control group that received distilled water (10 ml/kg) and three test groups which received 134 and 267 and 533 mg/kg of the aqueous extract of Pycnanthus angolensis daily per os. On the 29th day, the animals were killed, and several biochemical parameters were assessed for androgenic properties.&#x0D; Results: At a dose of 134 mg/kg, the extract significantly increased (p&lt;0.05) the levels of testicular cholesterol, testosterone and serum prostatic acid phosphatase activity compared to those of the normal control. A toxicity study showed that at doses of 134 and 267 mg/kg, the extract did not induce any significant variation (p&gt;0.05) in the creatinine level, but a significant reduction (p&lt;0.05) in of aspartate and alanine aminotransferase activities was recorded. Histopathology revealed destruction of testicular spermatozoa at a dose of 533 mg/kg compared to that of the controls.&#x0D; Conclusion: These results showed that at a dose of 134 mg/kg, the aqueous extract of Pycnanthus angolensis (Welw.) Warb. wood is capable of stimulating the production of androgens and is tolerated by the rat's body. Our results justified the traditional use of this plant for the treatment of male infertility and erectile dysfunction.

Abstract 3572: Role of the gut microbiome in androgen production and prostate cancer treatment resistance

Abstract Metastatic prostate cancer is often treated via a combination of androgen deprivation therapy (ADT) and the androgen receptor axis-targeted therapies abiraterone acetate + prednisone (AA/P) or enzalutamide. Unfortunately, most individuals undergoing these treatments develop resistance, termed “castration resistance”, through an unknown mechanism. Previous studies demonstrate a potential link between the gut microbiota and the treatment efficacy of endocrine therapy in metastatic castration resistant prostate cancer (mCRPC). This may be due in part to gut bacterial communities with the machinery to synthesize androgens using mechanisms distinct from human cells. For example, the bacterial desAB genes (e.g., “desmolase”) that were first described in the gut commensal bacterial species Clostridium scindens convert cortisol and prednisone to the androgenic metabolites 11β-hydroxyandrost-4-ene-3,17-dione (11OHAD) and ∆1-adrenosterone (∆1-AT), respectively. We hypothesize that androgen synthesis by the gut microbiota promotes treatment resistance to AA/P in advanced prostate cancer. This study aims to determine if androgen-converting gut bacterial species as well as circulating and fecal androgen levels correlate with AA/P treatment response in individuals with mCRPC using a combination of metagenomics and metabolomics. We further aim to demonstrate the relationship between gut bacterial androgen production and prostate cancer treatment response using an in vivo mouse model. Metagenomic sequencing showed an imbalance in microbial communities and identified the presence of androgen-synthesizing bacteria such as C. scindens in fecal samples of individuals undergoing treatment with AA/P. We also demonstrate that the relative abundance of C. scindens, as well as other species reported to generate androgens, are significantly enriched during metastatic disease progression on AA/P. Furthermore, targeted quantitative PCR of the bacterial desmolase (desA) gene demonstrated that absolute levels of desmolase are significantly higher in AA/P non-responders relative to AA/P responders, and trended higher in individuals with rising prostate-specific antigen (PSA) versus stable PSA while on AA/P. Targeted LC/MS/MS analyses of fecal samples demonstrated the presence of testosterone, dihydrotestosterone, 11OHAD, ∆1-AT, and other androgen metabolites in association with rising PSA levels in the AA/P cohort. Furthermore, we demonstrate that C. scindens converts hydrocortisone and prednisone into 11OHAD and ∆1-AT, respectively, in vitro at 48- and 72h post-treatment, which recapitulates prior published studies. Finally, we have established a prostate cancer mouse model using a VCaP xenograft in which tumor growth was significantly inhibited by abiraterone acetate in comparison to treatment with ADT alone. Additional studies will use this mouse model to study the effects of androgen metabolism by gut bacteria on AA/P efficacy. Overall, this study ascertains the ability of the human gut microbiota to harbor androgen-synthesizing bacteria, which in turn can be an alternative source of androgens that could impact the efficacy of the anti-androgen therapies used to treat metastatic prostate cancer. Citation Format: Angelica I. Cruz Lebron, Pedro A. Balbuena-Almodóvar, Luke Mummert, Sarah Ernst, Mark Markowski, Michelle Rudek, Jason Ridlon, Karen S. Sfanos. Role of the gut microbiome in androgen production and prostate cancer treatment resistance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3572.

(106) Stem Cell Therapy Leads to an Increase in Testosterone Levels in Infertile Men

Abstract Introduction Sexual dysfunction is frequently encountered in infertile males. This may be related to psychosexual aspects of infertility and to some common factors of pathogenesis, such as hypogonadism. For example, many patients with non-obstructive azoospermia (NOA) have low testosterone levels, which may lead to low sex drive, erectile dysfunction and metabolic abnormalities later in life. Patients with NOA typically undergo surgical sperm retrieval procedures, which may lead to further decrease in androgen production, which may be transient or even permanent in some cases. Stem cell therapy is an investigative mode of treatment for patients with NOA, but its influence on Leydig cell function is currently unknown. Objective To assess the impact of stem cell therapy on testosterone levels in men. Methods Sixty patients with NOA were included in this study. All of them underwent unsuccessful MicroTESE more than 12 months prior to the inclusion in order to justify the use of stem cell therapy. Multipotent mesenchymal stromal cells (MMSC) were derived from healthy donor placenta following physiologic childbirth. MMSC product was used for systemic and local treatment via intravenous and intratesticular injection, respectively. Endocrine profiling was performed at baseline and 3 months after the procedure. Results Median testosterone level prior to MMSC therapy was 8.7 nmol/l (interquartile range [IQR]: 4.2–12.1), and 3 months after it was 12.3 nmol/l (IQR: 8.8–16.3) (p &amp;lt; 0.00001). This may be related to an increase in luteinizing hormone levels from 6.4 mIU/ml at baseline (IQR: 3.6–11.3) to 9.6 mIU/ml (IQR: 5.2–14.1) (p = 0.00026). Paradoxically, follicle-stimulating hormone levels declined from 12.5 mIU/ml (IQR: 4.7–24.7) to 7.4 mIU/ml (IQR: 4.2–12.3) (p &amp;lt; 0.00001). Prior to MMSC therapy 43 patients (71.7%) had testosterone levels lower than 12 nmol/l, after MMSC therapy 24 patients (40%) had testosterone levels lower than this threshold. Thirty patients (50%) and 14 patients (23.3%) had testosterone level below 8 nmol/l before and after MMSC therapy respectively. Conclusions MMSC therapy may increase testosterone levels in patients with NOA, though the exact mechanisms and its possible impact on quality of life should be elucidated. Disclosure No.