Physiological Levels Of Testosterone Research Articles

Single-cell transcriptomics identifies senescence-associated secretory phenotype (SASP) features of testicular aging in human.

The male reproductive system experiences degradation with age, predominantly impacting the testes. Testicular aging can result in failure to produce physiological testosterone levels, normal sperm concentrations, or both. However, we cannot predict the onset of testicular aging in advance. Using single-cell RNA sequencing (scRNA-seq) from Gene Expression Omnibus (GEO) database, we conducted cell-cell communication network of human testis between older and young group, indicating Leydig cells' potential role in spermatogenesis microenvironment of aging testis. And we depicted the senescence-Associated Secretory Phenotype (SASP) features of aging testis by identifying differentially expressed senescence-associated secretory phenotype (SASP)-related genes between two group. Notably, IGFBP7 mainly expressed in Leydig cells of those differentially expressed SASP-related genes in aging testis. Furthermore, IGFBP7 protein located in the interstitial compartment of older mice confirmed by immunofluorescence and highly expressed in both human seminal plasma and mouse testis in the older group confirmed through Western blot. Together, our findings suggest that IGFBP7 may be a new biomarker of testicular aging.

(088) Impact of Testosterone Replacement Therapy on Adverse Cardiovascular Events in Hypogonadism Patients: A Comprehensive Evaluation of Physiological and Supraphysiological T Levels

Abstract Introduction Sustained treatment with elevated levels of exogenous testosterone (T) has been linked to various adverse events, particularly concerning cardiovascular risks. However, there is currently a lack of sufficient literature that comprehensively describes the overall safety profile of supraphysiologic testosterone levels. Objective To compare the safety profiles and rates of polycythemia, venous thromboembolism (VTE), major adverse cardiac events (MACE), deep vein thrombosis (DVT), myocardial infarction (MI), and stroke between hypogonadism patients treated with testosterone replacement therapy (TRT) at physiological and supraphysiological testosterone levels. Methods We identified patients with hypogonadism (<300 ng/dL) who underwent testosterone replacement therapy (TRT) at our institution through electronic medical record review. Patients lacking records of their first encounter, documented medical history, sufficient follow-up, or post-TRT adverse event data were excluded from the analysis. Supraphysiological testosterone levels were defined as >1000 ng/dL. The included patients were retrospectively analyzed using two-way nonparametric testing for continuous variables and chi-square and Fisher exact tests for categorical variables (α = 0.05). Results From a cohort of 537 men diagnosed with hypogonadism, we included 184 patients who underwent testosterone replacement therapy (TRT). Among these patients, 135 (73.4%) were treated to achieve physiological testosterone levels, with a median level of 468.0 ng/dL (interquartile range, IQR = 308.0-644.5), while 49 patients (26.6%) were treated to attain supraphysiologic testosterone levels, with a median level of 1552 ng/dL (IQR = 1279-1700). Before initiating TRT, there was no significant difference in testosterone levels between the two groups (p = 0.11). Moreover, no significant differences were found between the groups in terms of a history of diabetes mellitus (DM; p = 0.422) or prostate cancer (p = 0.29). However, significant differences were observed in the history of hypertension (HTN; p = 0.003) and hyperlipidemia (HLD; p = 0.006), with more patients in the physiologic testosterone group having a diagnosis of HTN and HLD prior to treatment. Regarding adverse events, there were more cases of polycythemia in the supraphysiologic testosterone group compared to the physiologic testosterone group (p < 0.001; 35.4% vs. 7.0%). Interestingly, there was no significant difference in the rate of other adverse events after TRT between the two groups, including venous thromboembolism (VTE; p = 0.285), major adverse cardiovascular events (MACE; p = 0.768), deep vein thrombosis (DVT; p > 0.999), myocardial infarction (MI; p = 0.562), and stroke (p > 0.999). Conclusions Despite widespread concerns, these findings suggest that although patients treated to achieve supraphysiologic testosterone levels had a higher incidence of polycythemia, there were no significant differences in the rates of other adverse events after TRT between patients treated to attain physiological versus supraphysiologic testosterone levels. Disclosure Any of the authors act as a consultant, employee or shareholder of an industry for: Consultant for AbbVie, Marius, Tolmar, Endo, Petros, Boston Scientific, Coloplast Investor: Sprout.

(009) Testosterone Management in Women with Hypoactive Sexual Desire Disorder Who Have Reduced 5α-Reductase Activity: A Unique Population

Abstract Introduction According to the Global Consensus Position Statement on the Use of Testosterone Therapy for Women, testosterone can significantly improve desire in menopausal women with hypoactive sexual desire disorder. Based on safety and efficacy of different formulations, the recommended treatment is 1/10th of a man’s dose, using a government approved topical testosterone gel or solution, or AndroFeme which is approved for use in menopausal women in Australia. There are, however, a subset of women without significant improvement in desire with this dose who experience improvement using higher doses of testosterone. These women have low or lower tertile dihydrotestosterone blood test values despite their total testosterone being above normal range or even in the upper tertile. The enzyme 5α-reductase converts testosterone to the more active form, dihydrotestosterone, that has a higher affinity for the androgen receptor than testosterone. 5α-reductase activity varies in different individuals. It is postulated that there are menopausal women with HSDD who have reduced 5α- reductase activity and therefore require a higher dose of testosterone from the gel or solution, or need a different testosterone formulation to realize clinical benefit. This subset of women do not appear to experience the usual adverse side effects of elevated testosterone (acne, thinning scalp hair, facial hair, aggressive behavior), in part because of low levels of dihydrotestosterone. Objectives This study examines the efficacy and safety of higher doses of testosterone using products government approved for men. Methods This is a retrospective chart review of menopausal women who were provided menopause management at our clinic focusing on women for whom 1/10th of a man’s dose of gel or solution did not provide symptom relief. Options included increasing the dose of gel or solution, or administration of an alternative delivery system of FDA approved (for men) testosterone including insertion of one or two 75 mg testosterone pellets (Testopel) or weekly intramuscular injection of testosterone cypionate (100 mg/ml – 0.1 ml or 10 mg). Results A total of 11 women (mean age 58 +/− 6 years) failed the usual 1/10th dose of topical testosterone gel for treatment of HSDD. Using testosterone pellet therapy, with 1-2 pellets implanted every 4-6 months, 6 patients had excellent desire as well as energy and self-confidence; 4 patients had excellent clinical response to weekly IM testosterone administration; 1 had improved symptoms by increasing the gel to 1 tube/4 days. While these treatments resulted in systemic levels of total testosterone outside the normal range for women (100 – 200 ng/dl), dihydrotestosterone values remained low or in the lower tertile (mean value 36 +/− 8 ng/dl; normal range 30 – 80 ng/dl). In general, side effects to be expected with higher doses of testosterone were not experienced despite elevated testosterone values. No patient complained of side effects consistent with excess testosterone other than mild facial hair. Conclusions In some women with reduced 5α-reductase activity, higher testosterone levels result in improved quality of life without expected negative side effects from higher physiologic levels of testosterone. Disclosure No.

Altered cardiac repolarization associated with cellular electrophysiological remodeling following chronic testosterone undecanoate administration in a canine model

Abstract Funding Acknowledgements Type of funding sources: None. Background The influence of testosterone on ventricular ion channels is widely investigated. Despite the legally stipulated availability of different androgen anabolic steroids (AAS), these agents are very popular among young adults, however, the direct effects of supra-physiological testosterone level are still unclear. Supposedly, the chronic use of AASs may result in structural and functional remodeling of the heart. Purpose The aim of our study was to investigate the potential electrophysiological effects of chronic administration of testosterone-undecanoate in a canine model in in vivo and in vitro studies. Methods Eight male beagle dogs were randomized into control (’Cont’) and treated (’Tr’) groups (n = 4; n = 4). The latter group received 15 mg/kg of long-lasting testosterone-undecanoate intramuscular injections weekly for 3 months. Blood samples were taken for monitoring testosterone levels. To investigate the altered repolarization in conscious dogs electrocardiography studies were performed. Ventricular myocytes were enzymatically dissociated via retrograde perfusion. The transmembrane ionic currents were recorded using the whole-cell configuration of the patch-clamp technique and the action potential duration (APD) was measured by the perforated patch-clamp technique. Results Testosterone level was significantly higher in the ‘Tr’ group compared to the ‘Cont’ group (47.02 nm/L vs. 15.23 nm/L; p=0.0002). The chronic treatment did not affect the heart rate between the examined groups (93.7±23.5 vs.108.8±21.4 beats/min), however, it resulted in significantly shortened QT (226±49 vs. 244.9±26.6 ms; p<0.05) and QTc (26.06±2.5 vs. 29.6±3.01 ms, p<0.05) intervals. ECG recordings also presented prolonged PQ (112.1±15.5 vs. 61.65±12.7ms; p<0.05), QRS (72.37±15.4 vs. 61.65±12.7 ms; p<0.05), and Tp-Te (32.95±7.45 vs. 53.46±16.6 ms; p<0.05) intervals in the ‘Tr’ group. Additionally, the APD of isolated left ventricular myocytes significantly shortened in the ‘Tr’ group compared to the ‘Con’ group (235.2±26.7 vs. 283.6±28.5 ms; p<0.05). Patch-clamp experiments revealed increased magnitude of transient outward potassium current (Ito), the inward rectifier potassium current (Ik1), and the slowed delayed rectifier potassium current (Iks) in the ‘Tr’ group. Conclusion The repolarization of the canine ventricular myocardium was significantly modified by constantly high level of testosterone. Many beneficial effects are attributed to physiological testosterone levels; however, the supra-physiological testosterone level may lead to potentially harmful alternations in the cardiac repolarization that may promote arrhythmogenesis, especially in the presence of various heart diseases. Presumably, the constantly high testosterone level induced electrophysiological changes may contribute to the development of life-threatening arrhythmias under certain circumstances.

Sexual attraction to visual sexual stimuli in association with steroid hormones across menstrual cycles and fertility treatment

BackgroundSteroid hormones (i.e., estradiol, progesterone, and testosterone) are considered to play a crucial role in the regulation of women’s sexual desire and sexual attraction to sexual stimuli throughout the menstrual cycle. However, the literature is inconsistent, and methodologically sound studies on the relationship between steroid hormones and women’s sexual attraction are rare. MethodsThis prospective longitudinal multisite study examined estradiol, progesterone, and testosterone serum levels in association with sexual attraction to visual sexual stimuli in naturally cycling women and in women undergoing fertility treatment (in vitro fertilization, IVF). Across ovarian stimulation of fertility treatment, estradiol reaches supraphysiological levels, while other ovarian hormones remain nearly stable. Ovarian stimulation hence offers a unique quasi-experimental model to study concentration-dependent effects of estradiol. Hormonal parameters and sexual attraction to visual sexual stimuli assessed with computerized visual analogue scales were collected at four time points per cycle, i.e., during the menstrual, preovulatory, mid-luteal, and premenstrual phases, across two consecutive menstrual cycles (n = 88 and n = 68 for the first and second cycle, respectively). Women undergoing fertility treatment (n = 44) were assessed twice, at the beginning and at the end of ovarian stimulation. Sexually explicit photographs served as visual sexual stimuli. ResultsIn naturally cycling women, sexual attraction to visual sexual stimuli did not vary consistently across two consecutive menstrual cycles. While in the first menstrual cycle sexual attraction to male bodies, couples kissing, and at intercourse varied significantly with a peak in the preovulatory phase, (all p ≤ 0.001), there was no significant variability across the second cycle. Univariable and multivariable models evaluating repeated cross-sectional relationships and intraindividual change scores revealed no consistent associations between estradiol, progesterone, and testosterone and sexual attraction to visual sexual stimuli throughout both menstrual cycles. Also, no significant association with any hormone was found when the data from both menstrual cycles were combined. In women undergoing ovarian stimulation of IVF, sexual attraction to visual sexual stimuli did not vary over time and was not associated with estradiol levels despite intraindividual changes in estradiol levels from 122.0 to 11,746.0 pmol/l with a mean (SD) of 3553.9 (2472.4) pmol/l. ConclusionsThese results imply that neither physiological levels of estradiol, progesterone, and testosterone in naturally cycling women nor supraphysiological levels of estradiol due to ovarian stimulation exert any relevant effect on women’s sexual attraction to visual sexual stimuli.

A Scoping Review of Transgender Policies in the 15 Most Commonly Played UK Professional Sports

There has been much debate recently on the participation of transgender and gender-diverse (TGD) athletes in sport, particularly in relation to fairness, safety and inclusion. The 2021 IOC Framework on Fairness, Inclusion and Non-discrimination acknowledges the central role that eligibility criteria play in ensuring fairness, particularly in the female category, and states that athletes should not be excluded solely on the basis of their TGD identity. To identify policies that address TGD athlete participation in the 15 major United Kingdom (UK) sporting organisations and to summarise the evidence for each of these policies. A scoping review of TGD policies from the 15 major UK sporting organisations. Eleven of the governing bodies had publicly available TGD policies. Most of the sporting associations drew guidance from the official 2015 IOC Consensus Meeting on Sex Reassignment and Hyperandrogenism, particularly with regard to physiological testosterone levels. Many organisations referenced their policies as a guide for decision making but stated that they ultimately made case-by-case decisions on an athlete's eligibility. Relevant considerations not addressed in most policies included pre- versus post-pubertal athletes, justification for testosterone thresholds, the length of time out of competitive action (if any) for transitioning athletes, the irreversible advantage from male puberty (if any), the responsibility for and frequency of follow up for hormonal testing and the consequences for athletes outside set testosterone limits. There is a lack of consensus among the top 15 UK sporting organizations relating to elite sport participation for TGD athletes. It would be useful for sport organizations to work together to develop greater standardization/consensus for TGD athlete policies, taking into consideration fairness, safety and inclusion in each sport.

Recovery of serum testosterone levels is an accurate predictor of survival from COVID-19 in male patients

BackgroundSARS-CoV-2 infection portends a broad range of outcomes, from a majority of asymptomatic cases to a lethal disease. Robust correlates of severe COVID-19 include old age, male sex, poverty, and co-morbidities such as obesity, diabetes, and cardiovascular disease. A precise knowledge of the molecular and biological mechanisms that may explain the association of severe disease with male sex is still lacking. Here, we analyzed the relationship of serum testosterone levels and the immune cell skewing with disease severity in male COVID-19 patients.MethodsBiochemical and hematological parameters of admission samples in 497 hospitalized male and female COVID-19 patients, analyzed for associations with outcome and sex. Longitudinal (in-hospital course) analyses of a subcohort of 114 male patients were analyzed for associations with outcome. Longitudinal analyses of immune populations by flow cytometry in 24 male patients were studied for associations with outcome.ResultsWe have found quantitative differences in biochemical predictors of disease outcome in male vs. female patients. Longitudinal analyses in a subcohort of male COVID-19 patients identified serum testosterone trajectories as the strongest predictor of survival (AUC of ROC = 92.8%, p < 0.0001) in these patients among all biochemical parameters studied, including single-point admission serum testosterone values. In lethal cases, longitudinal determinations of serum luteinizing hormone (LH) and androstenedione levels did not follow physiological feedback patterns. Failure to reinstate physiological testosterone levels was associated with evidence of impaired T helper differentiation and augmented circulating classical monocytes.ConclusionsRecovery or failure to reinstate testosterone levels is strongly associated with survival or death, respectively, from COVID-19 in male patients. Our data suggest an early inhibition of the central LH-androgen biosynthesis axis in a majority of patients, followed by full recovery in survivors or a peripheral failure in lethal cases. These observations are suggestive of a significant role of testosterone status in the immune responses to COVID-19 and warrant future experimental explorations of mechanistic relationships between testosterone status and SARS-CoV-2 infection outcomes, with potential prophylactic or therapeutic implications.

54. Adolescent Transgender Boys: Effects of 2 Years of Testosterone Therapy

An estimated 0.7% of the adolescent population experiences gender dysphoria. Some will seek gender-affirming hormone therapy. Transgender boys (those assigned female at birth who identify as male) take testosterone, typically as subcutaneous or intramuscular injections, to cause masculinization and achieve physiological levels of testosterone as seen in assigned males at birth. Testosterone may cause erythrocytosis. Current dosing guidelines are based on androgen therapy for adult hypogonadal cisgender men.

Effects of testosterone and exercise training on bone microstructure of rats.

Background and Aim:Male hypogonadism results from failure to produce physiological levels of testosterone. Testosterone in men is essential in masculine development, sperm production, and adult man’s health. Osteoporosis is one of the consequences of hypogonadism. Regular physical exercise and exogenous testosterone administration are frequently used to prevent or treat this condition. This study aimed to understand the effects of lifelong exercise training and testosterone levels (isolated and together) in the main bone structure parameters.Materials and Methods:A total of 24 rats were used and randomly divided into four groups: Control group (CG; n=6), exercised group (EG, n=6), testosterone group (TG, n=6), and testosterone EG (TEG, n=6). A micro-computed tomography equipment was used to evaluate 15 bone parameters.Results:Both factors (exercise training and testosterone) seem to improve the bone resistance and microstructure, although in different bone characteristics. Testosterone influenced trabecular structure parameters, namely, connectivity density, trabecular number, and trabecular space. The exercise promoted alterations in bone structure as well, although, in most cases, in different bone structure parameters as bone mineral density and medullar mineral density.Conclusion:Overall, exercise and testosterone therapy seems to have a synergistic contribution to the general bone structure and resistance. Further studies are warranted, comparing different individual factors, as gender, lifestyle, or testosterone protocols, to constantly improve the medical management of hypogonadism (and osteoporosis).

A new oral testosterone (TLANDO) treatment regimen without dose titration requirement for male hypogonadism.

BackgroundMale hypogonadism (testosterone level < 300 ng/dl) is a clinical syndrome that results from failure of the testis to produce physiological levels of testosterone. Most marketed testosterone replacement therapy products often require multiple dose adjustment clinic visits to achieve the desired, eugonadal testosterone levels.ObjectiveTo evaluate the efficacy and safety of a novel oral testosterone undecanoate therapy for the treatment of hypogonadism.Material and methodsNinety‐five (N = 95) hypogonadal men were enrolled in this open‐label, single‐arm, multicenter study in the United States (NCT03242590). Subjects received 225 mg of oral testosterone undecanoate (TLANDO) twice a day for 24 days without dose adjustment. Primary efficacy was percentages of subjects who achieved mean 24‐h testosterone levels within the eugonadal range and secondary efficacies were evaluated based on the upper limit of lab normal range of testosterone concentration.ResultsSubjects enrolled were on average age of 56 years, with about 17% of subjects older than 65 years. The mean body mass index was 32.8 kg/m2. The baseline mean total testosterone values were below the normal range (202 ± 74 ng/dl). Post‐treatment with 450 mg testosterone undecanoate daily dose without dose adjustment, 80% of subjects (95% confidence interval of 72%–88%) achieved a testosterone Cavg in the normal range and restored testosterone levels to mean testosterone Cavg of 476 ± 184 ng/dl at steady state. Testosterone restoration was comparable to other approved testosterone replacement therapy products. TLANDO was well tolerated with no deaths, no drug‐related serious adverse events, and no hepatic adverse events.Discussion and conclusionsTLANDO restored testosterone levels to the normal range in the majority of hypogonadal males. This new oral testosterone replacement therapy can provide an option for no‐titration oral testosterone replacement therapy. This therapy has the potential to improve patient compliance in testosterone replacement therapy.

Mice lacking the proton channel Hv1 exhibit sex-specific differences in glucose homeostasis

Sex as a physiologic factor has a strong association with the features of metabolic syndrome. Our previous study showed that loss of the voltage-gated proton channel Hv1 inhibits insulin secretion and leads to hyperglycemia and glucose intolerance in male mice. However, there are significant differences in blood glucose between male and female Hv1-knockout (KO) mice. Here, we investigated the differences in glucose metabolism and insulin sensitivity between male and female KO mice and how sex steroids contribute to these differences. We found that the fasting blood glucose in female KO mice was visibly lower than that in male KO mice, which was accompanied by hypotestosteronemia. KO mice in both sexes exhibited higher expression of gluconeogenesis-related genes in liver compared with WT mice. Also, the livers from KO males displayed a decrease in glycolysis-related gene expression and an increase in gluconeogenesis-related gene expression compared with KO females. Furthermore, exogenous testosterone supplementation decreased blood glucose levels in male KO mice, as well as enhancing insulin signaling. Taken together, our data demonstrate that knockout of Hv1 results in higher blood glucose levels in male than female mice, despite a decreased insulin secretion in both sexes. This sex-related difference in glucose homeostasis is associated with the glucose metabolism in liver tissue, likely due to the physiological levels of testosterone in KO male mice.

An oral lipidic native testosterone formulation that is absorbed independent of food.

ContextThere is no licensed oral native testosterone (NT) because of challenges in the formulation. Licensed oral formulations of the ester, testosterone undecanoate (TU), require a meal for absorption and generate supraphysiological dihydrotestosterone (DHT) levels.ObjectiveTo develop an oral NT formulation.Design and methodsA lipid-based formulation of native testosterone filled into soft-gelatin capsules at 40 mg per capsule was designed with 2 years of stability at ambient temperature. Pharmacokinetic comparison studies of this oral lipidic NT formulation to oral TU were conducted in dogs and hypogonadal men.ResultsIn dogs, 40 mg NT was well absorbed under fasted conditions whereas 40 mg TU required a high-fat meal: for NT, the mean fed/fasted AUC ratio was 1.63 and for TU 7.05. In hypogonadal men, fed and fasted NT had similar pharmacokinetics: Cmax mean 26.5 vs 30.4 nmol/L (769 vs 882 ng/dL), AUC0–10 h 87 vs 88.6 h nmol/L. NT (fed state) showed a testosterone AUC increase of 45% between 120 and 200 mg, and NT 200 mg gave a similar mean AUC0–10 h to TU 80 mg: 87 vs 64.8 h nmol/L. Serum TU levels were variable and on a molar basis were ~ten-fold higher than serum testosterone levels after TU 80 mg fed. The DHT: testosterone AUC0–10 h ratio was more physiological for NT than TU being 0.19 vs 0.36. There were no emerging safety concerns with NT.ConclusionThis novel oral lipidic native testosterone formulation has potential advantages over oral TU of dosing independently of food and a lower risk of supraphysiological DHT levels. Significance statement There is no licensed oral testosterone because of challenges in formulation, and the oral formulations of the ester, testosterone undecanoate, require a fatty meal for absorption and generate supraphysiological dihydrotestosterone levels. We have overcome the design challenges and formulated an oral native testosterone that can be taken with or without food and provides physiological levels of testosterone and dihydrotestosterone in hypogonadal men. This formulation, DITEST, has the potential advantage of being oral for patients who do not tolerate injections and less risk of adverse events that might theoretically be associated with elevated dihydrotestosterone levels. Future studies will need to define the dosing regimen for replacement in hypogonadal men.

Portomesenteric venous thrombosis in a postmenopausal female with testosterone implant: a case report

BackgroundAcute portal vein thrombosis is a rare medical event usually seen in liver disease, but it can also occur due to any inherited or acquired procoagulable state that triggers venous occlusion. Hormonal therapies have been associated with an increased risk of prothrombotic states. This case report documents a portomesenteric venous thrombosis in a postmenopausal woman with testosterone implant for the treatment of hypoactive sexual desire and discusses the importance of identifying hypercoagulable risk factors before initiating hormone replacement therapy. We want to improve the awareness of an unusual medical complication associated with hormone replacement therapy and shed light on how testosterone implants could facilitate a thrombotic event related to other risk factors such as obesity and chronic hypoxic states, as well as the importance of differential diagnosis in the evaluation of postmenopausal women on testosterone replacement therapy presenting with acute abdominal pain.Case presentationA 55-year-old obese postmenopausal Hispanic female with medical history of chronic obstructive pulmonary disease presents with intractable abdominal pain, is found to have elevated hemoglobin and hematocrit, and an abdominopelvic computed tomography scan revealing portal and superior mesenteric vein thrombosis. Further evaluation excluded inherited and acquired thrombophilia but revealed elevated testosterone levels. The patient was treated with anticoagulation, which resulted in recanalization of the portal and superior mesenteric veins.ConclusionSupraphysiologic levels of testosterone caused by testosterone implants as a treatment of hypoactive sexual desire in postmenopausal women can contribute to thrombotic events in the presence of additional prothrombotic risk factors. Therefore, testosterone therapy should include a thorough risk assessment for prothrombotic states, be tailored to patients’ physiologic testosterone levels, and have close follow-up with testosterone level monitoring.

Recovery of Serum Testosterone Levels is an Accurate Predictor of Survival From COVID-19 in Male Patients

Background: Infection with SARS-CoV-2 portends a broad range of outcomes, from a majority of asymptomatic cases or mild clinical courses to a lethal disease. Robust correlates of severe COVID-19 include old age, male sex, poverty and co-morbidities such as obesity, diabetes and cardiovascular disease. A precise knowledge is still lacking of the molecular and biological mechanisms that may explain the association of severe disease with male sex.Methods: Clinical biochemical and haematological parameters of admission samples in 249 hospitalized male and 247 female COVID-19 patients, analysed for associations with outcome and sex. Longitudinal analyses of a subcohort of 114 male patients for biochemical and haematological parameters, as well as testosterone, luteinizing hormone and androstenedione, analysed for associations with outcome. Longitudinal analysis of circulating immune populations by flow cytometry in 24 male patients, analysed for associations of specific subpopulations with outcome.Findings: There are quantitative differences in biochemical predictors of disease outcome in male vs. female patients. Longitudinal analyses in a subcohort of male COVID-19 patients indicate that testosterone trajectories are the strongest predictors (AUC of ROC = 92.8%, p Interpretation: Recovery or failure to reinstate testosterone levels is strongly associated with survival or death from COVID-19 in male patients. Our observations are suggestive of a significant role of testosterone status in the immune responses to COVID-19.Funding Information: Ministerio de Ciencia e Innovación (RTI2018-096055-B-I00), Consejo Superior de Investigaciones Científicas’ COVID-19 Research Fund (CSIC-COV19-006, CSIC-COV19-201), Agència de Gestió d’Ajuts Universitaris i de Recerca (2020PANDE00048 and 2017SGR 1411 GRC) and Plan Nacional de I+D (PID-107139RB-C21) and Instituto Nacional de la Salud Carlos III (PI18/00346 and COVID-19_00416).Declaration of Interests: Authors declare no conflict of interest.Ethics Approval Statement: protocol reviewed and approved by the Hospital Vall d’Hebron Institutional Review Board (Medical Research Ethics Committee, protocol number PR(AG)329-2020). Immunophenotyping studies of peripheral blood cells underwent a separate review and approval process (protocol number PR(AG)242/2020).

Testosterone increases bradykinin-induced relaxation in the coronary bed of hypertensive rats.

Physiological or supraphysiological levels of testosterone appear to be associated with the development of risk factors for cardiovascular diseases such as hypertension, as this hormone modulates the release of endothelial factors. However, its actions are still controversial, especially in the coronary circulation of hypertensive animals. This study was designed to assess the effects of testosterone treatment (T) on endothelium-dependent coronary vascular reactivity in orchiectomized SHR. The animals were divided into SHAM, orchiectomized (ORX), ORX+T and ORX+T+aromatase inhibitor (AI). All treatments lasted 15 days. Blood pressure (BP) was measured. Dose-response curves to bradykinin (BK) were constructed using the Langendorff technique, followed by inhibition of endothelium mediators (NO, prostanoids, EETs) and potassium channels. The intensity of eNOS, COX-1, COX-2, Akt, and gp91phox protein expression was quantified by Western blotting. BP was elevated in SHAM, ORX+T, and ORX+T+AI groups. However, we did not observe differences in the ORX group. Baseline coronary perfusion pressure (CPP) remained unaffected. Orchiectomy did not change the BK-induced relaxation compared to the SHAM group, whereas testosterone treatment increased it. This response was diminished in the absence of NO, prostanoids, and EETs in the SHAM and ORX groups, while in ORX+T group the relaxation was diminished only in the absence of NO and EETs. There was no difference in eNOS, COX-1, COX-2, and gp91phox protein expression, though Akt expression was increased in ORX and ORX+T groups. These results show that testosterone treatment can modulate endothelial function, especially in the coronary circulation under hypertension conditions, via NO and EETs pathways.

SAT-LB6 First Human Trial of an Oral Native Testosterone Shows Physiological Levels of Testosterone and Dht in Both Fasted and Fed State in Hypogonadal Men

Introduction: The prevalence of male hypogonadism is estimated to be 6% in the USA (1). Current therapies have limited acceptability: gels can be messy and risk inadvertent dosing of others; injections are painful; and oral testosterone undecanoate (TU) delivers variable testosterone levels, requires concurrent ingestion of a fatty meal and may produce supraphysiological dihydrotestosterone (DHT) levels. We present the first human trial of an oral native testosterone preparation formulated to deliver physiological levels of testosterone irrespective of food intake. Aim: To compare the pharmacokinetics of DITEST (Diurnal Ltd Cardiff, UK) to an oil based oral TU formulation (Andriol Testocaps MSD, UK) and explore the effect of food on DITEST bioavailability. Methods: Single centre, phase 1b study of DITEST in 25 adult males with hypogonadism, one subject withdrawn after single period and only included in safety analysis (Clinicaltrials.gov: NCT02966652). Part 1 compared the pharmacokinetics of 80mg TU with 120mg DITEST after a high fat meal. Part 2 the pharmacokinetics of 200mg of DITEST administered in either fed or fasted states. Results are baseline adjusted. Results: DITEST showed a testosterone dose response between 120mg and 200mg with Cmax 550 (19.1) and 877 (30.4) ng/dl (nmol/l) and AUC0-10h 59.5 and 88.6 h*nmol/L. DITEST 200mg gave an equivalent Cmax and AUC0-10h to TU 80mg: Cmax 877 (30.4) vs 906 (31.4) ng/dl (nmol/l) and AUC0-10h 88.6 vs 102 h*nmol/L. Fed and fasted DITEST had similar pharmacokinetics: Cmax 764 (26.5) vs 877 (30.4) ng/dl (nmol/L), AUC0-10h 87.0 vs 88.6. DITEST resulted in lower levels of DHT than TU: Cmax 84 (2.9), 131 (4.5) & 194 (6.7) ng/dl (nmol/l); AUC0-10h 11.0, 16.7 & 36.3 h*nmol/L for DITEST 120mg, 200mg & 80mg TU, respectively. There was one serious adverse event (urinary retention) in the study during TU dosing. There were no emerging safety concerns, and adverse event frequency and severity was similar between the two treatments. Discussion: These results demonstrate that 200mg DITEST provides similar testosterone exposure with more physiological DHT exposure than 80mg TU given with a high fat meal. Administration of DITEST in fed and fasted states provides similar testosterone and DHT exposure. Compared to published literature on a self-emulsifying formulation of TU at 200mg, DITEST at 200mg provides a similar testosterone Cmax and no requirement for a fatty meal (2). Conclusion: DITEST is an oral native testosterone formulation with anticipated advantages over current oral therapy of dosing without food and a lower risk of supraphysiological DHT levels.

Scaffold-Free Endometrial Organoids Respond to Excess Androgens Associated With Polycystic Ovarian Syndrome.

Polycystic ovary syndrome (PCOS) is a prevalent disorder in reproductive aged women associated with a number of endocrine and metabolic complications, including increased risk of endometrial cancer. To study the effect of the characteristic increased androgen levels in PCOS on the endometrium, a novel scaffold-free multicellular endometrial organoid was established. Human endometrial organoids were constructed using primary endometrial epithelial and stromal cells from endometrial tissues. Organoids were treated for 14 days with physiologic levels of estradiol and testosterone to mimic a normal follicular phase or PCOS hormone profiles. Organoids were harvested for immunostaining and ribonucleic acid sequencing. Academic institution. Endometrial tissues from 10 premenopausal women undergoing hysterectomy for benign pathologies were obtained following written consent. Organoid architecture, cell specific markers, functional markers, proliferation, and gene expression were measured. A method to generate scaffold-free endometrial organoids containing epithelial and stromal cells was established. These organoids exhibited distinct organization with epithelial cells lining the outer surface and stromal cells in the center of the organoids. Epithelial cells were polarized, organoids expressed cell type specific and functional markers, as well as androgen, estrogen, and progesterone receptors. Treatment with PCOS hormones increased cell proliferation and dysregulated genes in endometrial organoids. A new multicellular, scaffold-free endometrial organoid system was established that resembled physiology of the native endometrium. Excess androgens in PCOS promoted cell proliferation in endometrial organoids, revealing new mechanisms of PCOS-associated with risk of endometrial neoplasia.

Generation of Multicellular Human Primary Endometrial Organoids.

The human endometrium is one of the most hormonally responsive tissues in the body and is essential for the establishment of pregnancy. This tissue can also become diseased and cause morbidity and even death. Model systems to study human endometrial biology have been limited to in vitro culture systems of single cell types. In addition, the epithelial cells, one of the major cell types of the endometrium, do not propagate well or retain their physiological traits in culture, and thus our understanding of endometrial biology remains limited. We have generated, for the first time, endometrial organoids that consist of both epithelial and stromal cells of the human endometrium. These organoids do not require any exogenous scaffold materials and specifically organize so that epithelial cells encompass the spheroid-like structure and become polarized with stromal cells in the center that produce and secrete collagen. Estrogen, progesterone and androgen receptors are expressed in the epithelial and stromal cells and treatment with physiological levels of estrogen and testosterone promote the organization of the organoids. This new model system can be used to study normal endometrial biology and disease in ways that were not possible before.

SUN-022 Absence of Neuronal Androgen Receptor (AR) Improves Glucose Homeostasis in Female Mice

Androgen excess predisposes females to type 2 diabetes (1). Using mouse models of testosterone excess, we reported that this is due, at least partially, to excess AR activation in pancreatic β-cells and neurons. Excess AR activation in β-cells causes β-cell dysfunction while excess AR activation in neurons causes peripheral insulin resistance (2). However, the role of neuronal AR in insulin sensitivity and glucose homeostasis in female mice under physiological testosterone levels is unknown. To address this issue, we used Cre-lox technology to generate female mice with neuronal androgen receptor knockout (NARKO). We explored the role of neuronal AR in glucose homeostasis in NARKO and littermate control female mice challenged with a western diet high in fat and sugars. At 2 months of age, female NARKO mice exhibited similar glucose homeostasis to that of control littermates. However, starting at 4 months of age, female NARKO mice developed improved glucose homeostasis. Compared to control littermates, female NARKO mice exhibited decreased glucose and insulin levels in the fasting state and following a glucose challenge. This phenotype became more pronounced at 6 and 8 months of age. These results suggest that the absence of AR signaling in neurons protects female mice challenged with nutrient excess from the development of insulin resistance and type 2 diabetes. Further studies are needed to determine the mechanism through which absence of neuronal AR protects female mice from insulin resistance.

Physiological increase of yolk testosterone level does not affect oxidative status and telomere length in gull hatchlings.

Conditions experienced during early-life can cause the onset of oxidative stress, resulting in pervasive effects on diverse life-history traits, including lifespan. In birds, maternally-transferred egg substances may exert positive or negative influence over the offspring phenotype. Among these, testosterone can upregulate the bioavailability of certain antioxidants but simultaneously promotes the production of pro-oxidants, leading to an oxidative stress situation, which is one of the main forces causing telomere attrition However, no study has investigated the role of this androgen on telomere dynamics in birds and little is known about the effects of yolk testosterone on oxidative status in early-life of these species. We physiologically increased the levels of yolk testosterone by in ovo injections in yellow-legged gull (Larus michahellis) to evaluate the effects induced by this androgen on hatchlings plasma total antioxidant capacity, amount of pro-oxidant molecules and telomere length at hatching. Testosterone supplementation did not increase hatchling body growth, did not result in the overproduction of pro-oxidant molecules nor a reduction of antioxidant capacity. Accordingly, telomere length at hatching was not affected by testosterone treatment, although hatchlings from the third-laid eggs showed shorter telomeres than their siblings from first- and second-laid eggs, independently of testosterone treatment. Our results suggest that injection of physiological levels of testosterone does not induce oxidative stress to hatchlings and, consequently do not affect telomere dynamics during early post-natal periods.