Androgen-dependent Disease Research Articles

Evaluation of glucocorticoid-related genes reveals GPD1 as a therapeutic target and regulator of sphingosine 1-phosphate metabolism in CRPC

Prostate cancer (PCa) is an androgen-dependent disease, with castration-resistant prostate cancer (CRPC) being an advanced stage that no longer responds to androgen deprivation therapy (ADT). Mounting evidence suggests that glucocorticoid receptors (GR) confer resistance to ADT in CRPC patients by bypassing androgen receptor (AR) blockade. GR, as a novel therapeutic target in CRPC, has attracted substantial attention worldwide. This study utilized bioinformatic analysis of publicly available CRPC single-cell data to develop a consensus glucocorticoid-related signature (Glu-sig) that can serve as an independent predictor for relapse-free survival. Our results revealed that the signature demonstrated consistent and robust performance across seven publicly accessible datasets and an internal cohort. Furthermore, our findings demonstrated that glycerol-3-phosphate dehydrogenase 1 (GPD1) in Glu-sig can significantly promote CRPC progression by mediating the cell cycle pathway. Additionally, GPD1 was shown to be regulated by GR, with the GR antagonist mifepristone enhancing the anti-tumorigenic effects of GPD1 in CRPC cells. Mechanistically, targeting GPD1 induced the production of sphingosine 1-phosphate (S1P) and enhanced histone acetylation, thereby inducing the transcription of p21 that involved in cell cycle regulation. In conclusion, Glu-sig could serve as a robust and promising tool to improve the clinical outcomes of PCa patients, and modulating the GR/GPD1 axis that promotes tumor growth may be a promising approach for delaying CRPC progression.

Evaluating treatments for male androgenetic alopecia: clinical profile and comparative efficacy of platelet rich plasma and minoxidil with finasteride

Background: Androgenetic alopecia (AGA) is a common, hereditary, and androgen-dependent disorder characterized by progressive hair thinning on the scalp, affecting both men and women. It arises from follicular miniaturization, leading to the conversion of terminal hair into vellus hair. Methods: This study is a single-center, prospective, open-label, randomized, placebo-controlled trial with four parallel arms designed to evaluate treatments for male pattern baldness. Eighty patients were randomized into four groups: Group 1 received topical minoxidil and finasteride solution, group 2 received PRP alone, group 3 received PRP with minoxidil and finasteride solution, and group 4 received normal saline (NS). Interventions were administered monthly over three months, followed by a three-month follow-up period. Treatment effects were assessed using measures such as patient self-assessment, global photography, and the hair pull test. Results: Group 3 (Combined treatment) showed the most significant improvement in the hair pull test after six months (73.33% negative tests), followed by group 2 (PRP treatment) with 60% improvement. Patient satisfaction scores were highest in group 3 across all follow-ups (F1 to F5), demonstrating superior treatment response compared to other groups (p<0.001). Global photography assessments indicated varying results among groups, with group 4 recording the highest proportion of fair improvements (58.5%). Group 3 exhibited the highest percentage of patients with above-average improvement. Conclusions: Our study provides valuable insights into managing AGA, emphasizing the effectiveness of combining PRP with topical treatments compared to individual therapies and placebo.

Abstract 7116: Targeting the CBP/p300 axis in lethal prostate cancer impacts DNA repair

Abstract Prostate cancer (PCa) is the 2nd leading cause of cancer related deaths in US men. PCa is an androgen dependent disease driven by the androgen receptor (AR). Currently, androgen-deprivation therapy (ADT) is the standard of care first-line therapy for metastatic PCa. Resistance to ADT uniformly leads to lethal disease, termed castration-resistant prostate cancer (CRPC). Thus, there is an unmet clinical need to identify and develop novel strategies to treat CRPC. CBP/p300 are potent co-activators for AR. High expression of CBP/p300 is associated with locally advanced disease and castration resistant function of AR, resulting in poor patient outcomes and further highlighting the need to discern the role of CBP/p300 to potentially develop therapeutic targets for precision medicine. Previous studies have relied on non-specific compounds and genetic silencing to target CBP/p300. In this study, CBP/p300 mediated bromodomain activity is targeted by CCS1477 (inobrodib), a first-in-class bromodomain inhibitor developed by Cell Centric. CCS1477 treatment demonstrated effective inhibition in growth and clonogenicity assays. Inhibition of the CBP/p300 bromodomain with CCS1477 resulted in significant downregulation of AR-FL, AR-V7, and its targets’ mRNA expression in addition to inhibition of associated factors such as c-MYC and its downstream targets in PCa cell lines as well as patient derived xenograft (PDX) models. This study shows that CBP and p300 are highly expressed and correlate closely with AR gene expression and AR activity score in primary PCa and CRPC patient samples. The clinical significance of CBP/p300 expression in PCa has been investigated via elucidation of CBP/p300 transcriptional reprogramming and its role in DNA damage response (DDR) pathways. Specifically, findings revealed that CBP/p300 bromodomain suppression sensitizes to AR-dependent DNA-repair. Transcriptional mapping identified CBP/p300 as regulators of cell proliferation and DNA repair processes, which were functionally confirmed across several PCa model systems. To assess relevance, exogenous challenge with radiation revealed that CBP/p300 are required for AR-mediated DNA repair, and CBP/p300 expression is linked to DNA repair capacity in the clinical setting. Molecular analyses revealed that CBP/p300 facilitate double-strand break (DSB) repair efficiency via homologous recombination (HR) mediated DDR. Congruently, CBP/p300 strongly correlated with HR gene expression in PCa patient tissue. These collective findings reveal that CBP/p300 govern repair of DNA DSBs by regulating HR, thus modulating genome integrity and promoting CRPC growth. These studies identify CBP/p300 as a driver of PCa tumorigenesis through coordinated control of critical transcriptional events and lay the groundwork to optimize therapeutic strategies for advanced PCa via CBP/p300 inhibition, potentially in combination with AR-directed therapies. Citation Format: Sumaira Sardar, Lakshmi Ravindranath, Christopher McNair, Saswati N. Chand, Wei Yuan, Denisa Bogdan, Jonathan Welti, Adam Sharp, Matthew Schiewer, Lisa Butler, Johann de Bono, Kris Frese, Nigel Brooks, Neil Pegg, Karen Knudsen, Ayesha A. Shafi. Targeting the CBP/p300 axis in lethal prostate cancer impacts DNA repair [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 7116.

Abstract B009: Targeting the AR co-activator CBP/p300 axis in castration resistant prostate cancer Impacts DNA damage repair function

Abstract Prostate cancer (PCa) is the second leading cause of cancer related deaths in US men. PCa is an androgen dependent disease and the transcription factor, androgen receptor (AR), is crucial for the overall outcome. Currently, androgen-deprivation therapy (ADT) is the standard of care first-line therapy for metastatic PCa. Resistance to ADT almost uniformly leads to lethal disease, termed castration-resistant prostate cancer (CRPC). Thus, there is a significant unmet clinical need to identify and develop novel strategies to treat CRPC. CBP/p300 are potent co-activators for AR. High expression of CBP/p300 are associated with locally advanced disease and castration resistant function of AR; thus, resulting in poor patient outcome and further highlighting the need to discern the role of CBP/p300 to potentially develop novel therapeutic targets for precision medicine. Previous studies have relied on non-specific compounds and genetic silencing to target CBP/p300 and its associated transcriptional machinery. In this study, CBP/p300 mediated bromodomain activity is targeted by CCS1477 (Inobrodib), a first-in-class bromodomain inhibitor developed by Cell Centric. CCS1477 was shown to demonstrate effective inhibition in growth and clonogenicity assays. Inhibition of the CBP/p300 bromodomain with CCS1477 resulted in significant downregulation of AR-FL, AR-V7, and its targets’ mRNA expression in addition to inhibition of associated factors such as c-MYC and its downstream targets in PCa cell lines as well as patient derived xenograft (PDX) models. This study shows that CBP and p300 are highly expressed and correlate closely with AR gene expression and AR activity score in primary PCa and CRPC patient samples. The clinical significance of CBP/p300 expression in PCa has been investigated via elucidation of CBP/p300 transcriptional reprogramming and role in DNA damage response (DDR) pathways. Specifically, findings revealed that CBP/p300 bromodomain suppression sensitizes to AR-dependent DNA-repair. Transcriptional mapping identified CBP/p300 as regulators of cell proliferation and DNA repair processes, which were functionally confirmed across several PCa model systems. To assess relevance, exogenous challenge with radiation revealed that CBP/p300 bromodomain is required for AR-mediated DNA repair, and CBP/p300 expression is linked to DNA repair capacity in the clinical setting. Molecular analyses revealed that CBP/p300 facilitate double-strand break (DSB) repair efficiency via homologous recombination (HR) mediated DDR. Congruently, CBP/p300 strongly correlated with HR gene expression in PCa patient tissue. These collective findings reveal that CBP/p300 govern repair of DNA DSBs by regulating HR, thus modulating genome integrity, and promoting CRPC growth. In conclusion, these studies identify CBP/p300 as a driver of PCa tumorigenesis through coordinated control of critical transcriptional events and lay the groundwork to optimize therapeutic strategies for advanced PCa via CBP/p300 inhibition, potentially in combination with AR-directed therapies. Citation Format: Sumaira Sardar. Targeting the AR co-activator CBP/p300 axis in castration resistant prostate cancer Impacts DNA damage repair function [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: DNA Damage Repair: From Basic Science to Future Clinical Application; 2024 Jan 9-11; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2024;84(1 Suppl):Abstract nr B009.

Androgenetic alopecia: evaluation of hormonal profile and its systemic implications

Background: Androgenetic Alopecia is a hereditary androgen-dependent disorder characterized by a gradual conversion of terminal hair into miniaturized hair with typical bitemporal recession and balding vertex and is considered the most common type of baldness characterized by progressive hair loss. This study evaluated the hormonal profile in males with androgenetic alopecia. This study evaluated the hormonal profile of early androgenetic alopecia in males. Methods: This prospective study included 84 patients attending the outpatient Department of Dermatology. Forty-four cases and 40 controls were included in the study. The study had 44 male patients presenting with complaints of grade ≥ 3 androgenetic alopecia in the age group 19-35 years, whereas 40 age and sex-matched patients attending Dermatology OPD for unrelated complaints with no history of hair loss or clinical examination suggestive of androgenetic alopecia were included in the control group. After a detailed history, and examination, testosterone, LH, FSH, Prolactin, and SHBG were estimated. Results: The mean age of onset was found to be 24.29±3.28 years. Positive family history was seen in 65.90% of patients. The mean testosterone, LH, FSH, prolactin, SHBG and free androgen index in cases versus controls was 6.44±2.58 versus 3.32±1.53 ng/ml, 8.01±2.64 IU/l versus 3.01±1.16 IU/l, 3.82±1.33 IU/l versus 5.07±1.27 IU/l, 15.50±5.11 ng/ml versus 9.84±3.91 ng/ml, 12.72±2.63 nmol/l versus 29.18±4.90 nmol/l and 51.03±21.78 versus 11.40±4.66 respectively. LH/FSH ratio was 2.17±0.54 versus 0.63±0.27. These parameters had p values <0.05 and were statistically significant. Conclusions: Our study concluded that serum testosterone, prolactin, LH, LH/FSH, and FAI are increased whereas serum FSH and SHBG are decreased in cases of androgenetic alopecia compared to controls.

A GATA2-CDC6 axis modulates androgen receptor blockade-induced senescence in prostate cancer

BackgroundProstate cancer is a major cause of cancer morbidity and mortality in men worldwide. Androgen deprivation therapy (ADT) has proven effective in early-stage androgen-sensitive disease, but prostate cancer gradually develops into an androgen-resistant metastatic state in the vast majority of patients. According to our oncogene-induced model for cancer development, senescence is a major tumor progression barrier. However, whether senescence is implicated in the progression of early-stage androgen-sensitive to highly aggressive castration-resistant prostate cancer (CRPC) remains poorly addressed.MethodsAndrogen-dependent (LNCaP) and –independent (C4-2B and PC-3) cells were treated or not with enzalutamide, an Androgen Receptor (AR) inhibitor. RNA sequencing and pathway analyses were carried out in LNCaP cells to identify potential senescence regulators upon treatment. Assessment of the invasive potential of cells and senescence status following enzalutamide treatment and/or RNAi-mediated silencing of selected targets was performed in all cell lines, complemented by bioinformatics analyses on a wide range of in vitro and in vivo datasets. Key observations were validated in LNCaP and C4-2B mouse xenografts. Senescence induction was assessed by state-of-the-art GL13 staining by immunocytochemistry and confocal microscopy.ResultsWe demonstrate that enzalutamide treatment induces senescence in androgen-sensitive cells via reduction of the replication licensing factor CDC6. Mechanistically, we show that CDC6 downregulation is mediated through endogenous activation of the GATA2 transcription factor functioning as a CDC6 repressor. Intriguingly, GATA2 levels decrease in enzalutamide-resistant cells, leading to CDC6 stabilization accompanied by activation of Epithelial-To-Mesenchymal Transition (EMT) markers and absence of senescence. We show that CDC6 loss is sufficient to reverse oncogenic features and induce senescence regardless of treatment responsiveness, thereby identifying CDC6 as a critical determinant of prostate cancer progression.ConclusionsWe identify a key GATA2-CDC6 signaling axis which is reciprocally regulated in enzalutamide-sensitive and -resistant prostate cancer environments. Upon acquired resistance, GATA2 repression leads to CDC6 stabilization, with detrimental effects in disease progression through exacerbation of EMT and abrogation of senescence. However, bypassing the GATA2-CDC6 axis by direct inhibition of CDC6 reverses oncogenic features and establishes senescence, thereby offering a therapeutic window even after acquiring resistance to therapy.

Abstract B058: Exploring the oncogenic role of six-transmembrane epithelial antigen of the prostate in the context of androgen receptor signaling

Abstract Prostate cancer is the second leading cause of death among American men, where pathogenesis is predominantly driven by dysregulation of the Androgen Receptor (AR). The current standard of care for metastatic disease includes a regime of androgen deprivation therapy (ADT) and AR antagonists, which increases survival to 24-48 months. Selective pressure of the therapy often leads to resurgence of AR activity resulting in castrate resistant disease, for which there are no curative treatment options. Moreover, once the disease has progressed to castrate resistance, AR directed therapies are no longer viable as a sole treatment option. Six-Transmembrane Epithelial Antigen of the Prostate 1-4 (STEAP) proteins are a family of ferric metalloreductases involved in maintaining iron homeostasis. STEAP expression is enriched in androgen sensitive and hormone refractory disease states, where increased expression correlates with high Gleason Score. STEAP proteins display greater intratumoral homogeneity at both primary and metastatic sites than the canonically used Prostate Specific Membrane Antigen (PSMA), indicating attractive diagnostic and therapeutic potential. Differential expression profiles have been observed among the STEAP family members depending on AR status in-vitro, suggesting a dependence on the AR signaling network. Previous literature indicates silencing of STEAPs in-vitro decreased cell viability and reduced proliferative, invasive, and migratory phenotypes; moreover in-vivo xenografts in which STEAPs are knocked down exhibited a significant reduction in tumor growth. Preliminary data suggests that treatment with Enzalutamide reduces the expression of STEAP proteins, implying a mechanistic link to the AR signaling axis. AMG509, a bispecific STEAP1-CD3 antibody, is the only STEAP directed therapeutic actively in clinical trials for prostate cancer (NCT04221542). There is limited mechanistic data regarding how STEAP directed therapies exert their anti-tumor function. Despite sharing similar architecture, localization, and biological implications, each STEAP protein retains unique functional roles within the cell. We plan to perform a systematic investigation on the oncogenic roles of individual STEAP protein in the context of AR dependent and independent prostate cancer models. In-vitro pharmacological AR manipulations will be used to evaluate changes in STEAP expression in multiple disease models. Genetic manipulations of STEAPs will be carried out under normal growth and ADT conditions to investigate the effects on growth, migration, oxidative stress, and cell cycle regulation. Given their principal role as ferric reductases, iron supplementation and iron drop-out will be examined in parallel. Novel treatment regimens will be explored using a combination of AR and STEAP directed therapeutics. Collectively, this study aims to uncover both the overlapping and distinct functions of STEAP proteins within the AR signaling axis and to illuminate their usefulness as a therapeutic target. Citation Format: Candice Bizzaro, Latese Evens, Moriah Cunningham, Jasibel Vasquez Gonzalez, Matthew Schiewer. Exploring the oncogenic role of six-transmembrane epithelial antigen of the prostate in the context of androgen receptor signaling [abstract]. In: Proceedings of the AACR Special Conference: Advances in Prostate Cancer Research; 2023 Mar 15-18; Denver, Colorado. Philadelphia (PA): AACR; Cancer Res 2023;83(11 Suppl):Abstract nr B058.

Abstract A077: MicroRNA drivers of resistance to androgen deprivation therapy in prostate cancer

Abstract INTRODUCTION: Prostate cancer is the most prevalent malignancy affecting Western males. It is initially an androgen-dependent disease: androgens bind to the androgen receptor and drive expression of genes that promote proliferation and evasion of apoptosis. Despite reduced androgen dependence in advanced prostate cancer, androgen receptor signalling remains a key driver of growth. Androgen deprivation therapy (ADT) is therefore a first line treatment approach and works well initially, but resistance inevitably develops. Abiraterone and Enzalutamide are drugs widely used in ADT and are androgen synthesis and androgen receptor signalling inhibitors respectively. The shortage of other treatment options means acquired resistance to these drugs is a major clinical problem. MicroRNAs (miRs) are important mediators of post-transcriptional gene regulation and show altered expression in cancer. Several have been linked to the development of resistance to ADT. Manipulation of such miRs may be a pathway to breakthrough treatments for advanced prostate cancer. This study aimed to validate ADT resistance-implicated miRs and their clinically relevant targets. MATERIALS AND METHODS: Small RNA-sequencing of Abiraterone- and Enzalutamide-resistant C42 prostate cancer cells identified subsets of miRs dysregulated as compared to parental cells. Real-Time Quantitative Reverse Transcription PCR (qRT-PCR) was used to validate altered expression of candidate ADT resistance-implicated miRs 195-5p, 497-5p and 29a-5p in ADT-resistant and -responsive prostate cancer cell lines, patient-derived xenografts (PDXs) and primary prostate cancer explants. RESULTS AND DISCUSSION: This study suggests a possible role for miR-497-5p in the development of ADT resistance in prostate cancer. MiR-497-5p expression was increased in ADT-resistant versus ADT-responsive prostate cancer cells. Importantly, miR-497-5p expression was also increased in Enzalutamide-treated, castrated (ADT-mimicking) PDXs versus intact PDXs. MiR-195-5p was also elevated in ADT-resistant versus -responsive prostate cancer cells, while there was a drop in miR-29a-5p expression. Candidate clinically relevant targets of miR-497-5p in prostate cancer were identified by mining AGO-PAR-CLIP-seq data sets and may include AVL9 and FZD6. CONCLUSION: In summary, this study identified microRNAs that are implicated in prostate cancer resistance to androgen deprivation therapy and could represent novel therapeutic targets for advanced disease. Citation Format: Philippa C. Saunders, Claire Fletcher. MicroRNA drivers of resistance to androgen deprivation therapy in prostate cancer [abstract]. In: Proceedings of the AACR Special Conference: Advances in Prostate Cancer Research; 2023 Mar 15-18; Denver, Colorado. Philadelphia (PA): AACR; Cancer Res 2023;83(11 Suppl):Abstract nr A077.

SERUM VITAMIN D LEVELS IN PREMATURE ANDROGENETIC ALOPECIA- A CASE -CONTROL STUDY.

Background: Androgenetic alopecia is a hereditary, androgen-dependent disorder characterized by a gradual conversion of terminal hair into miniaturized hair and causes progressive hair loss. Studies have showed lowered vitamin D levels in patients with androgenetic alopecia, which also correlated to their severity. The purpose of this study is to determine the association and correlation of Serum vitamin D levels in premature onset of androgenetic alopecia Methods and methodology: A total of fty males clinically diagnosed with androgenetic alopecia and fty age matched healthy controls were recruited. Serum Vitamin D levels of the cases and controls obtained were assessed. The data was analysed statistically. Results: In this study the mean levels of serum vitamin D were signicantly decreased in cases, compared to controls (13.90 vs. 28.4 nmol/L; P ≤ 0.001). Eighty seven percent of the cases had deciency of vitamin D (<25 nmol/L), while 13% had insufcient vitamin D levels (25-75 nmol/L). However, there was no correlation between severity of androgenetic alopecia and serum vitamin D levels. (P value =1.65). Conclusion: This study shows that majority of cases with early onset androgenetic alopecia had deciency of vitamin D levels. However, there was no correlation between severity of androgenetic alopecia and serum vitamin D levels

METABOLIC SYNDROME IN PATIENTS WITH ANDROGENETIC ALOPECIA IN A TERTIARY CARE HOSPITAL

Background: Androgenetic alopecia (AGA) is a hereditary androgen-dependent disorder, characterized by a progressive decline in hair fibre production by scalp hair follicles and their eventual miniaturization. Metabolic syndrome is a cluster of inter-related risk factors that increase the risk of coronary artery disease. Despite the high burden of Androgenetic Alopecia and Metabolic Syndrome in India, specific data on the participants are relatively sparse.
 
 Methods: A total of 126 patients of androgenetic alopecia (age range 18-55 years; mean age 30.83±10.37 years; 83.3% males) falling in sampling frame were enrolled in the study and were assessed clinically, anthropometrically and biochemically. Metabolic syndrome was diagnosed using Joint interim statement of International Diabetes Federation, National Heart, Lung and Blood Institute, American Heart Association, World Health Federation, International Atherosclerosis Society, and International Association of the Study of Obesity criteria.
 
 Results: Prevalence of Metabolic Syndrome (MetS) was 29.4% in 126 cases of Androgenetic Alopecia. The P Value was <0.001 for obesity (52.4%), hypertension (35.7%), low HDL cholesterol (21.4%), hypertriglyceridemia (13.5%) and high fasting glucose (12.7%) respectively. Patients with higher severity grade of AGA had significantly higher prevalence of MetS and its components.
 
 Conclusions: Metabolic Syndrome showed a positive relation with Androgenetic Alopecia. Early screening for Metabolic Syndrome is beneficial in patients with androgenic alopecia to prevent them from developing coronary artery disease.

Effect of equal access to care on prostate cancer progression in minority patients.

33 Background: Prostate cancer (PC) is an androgen-dependent disease that initially responds to androgen deprivation therapy (ADT); however, it can become resistant to therapies that reduce testosterone levels, signaling a transition to non-metastatic, castration-resistant PC (nmCRPC). Racial disparities are known in PC, with black and Hispanic men presenting with more advanced disease and having a higher mortality rate compared to non-Hispanic white men. The goal of treatment management in patients with nmCRPC is to prevent or delay metastases. This study examines the disease progression observed among patients with nmCRPC who received care and treatment in an equal access healthcare system, the Veterans Health Administration. Methods: Patient information was obtained from the Veterans Affairs (VA) Clinical Cancer Registry (VACCR) and the VA Corporate Data Warehouse (CDW) to identify patients who were diagnosed with prostate cancer at the VA and later developed nmCRPC, defined as (1) evidence of a rising prostate-specific antigen (PSA), i.e., 2 consecutive increases in PSA concentration over a baseline value; (2) evidence of ongoing androgen deprivation consisting of a serum testosterone level of ≤50 ng/dL (≤2.0 nmol/L); and (3) the absence of metastatic disease as evidenced in radiology reports. Treatments of nmCRPC were identified by review of NCCN Prostate Cancer Guidelines and extracted from CDW pharmacy dispensation records and indexed at nmCRPC diagnosis. A multivariate Cox proportional hazard model was used to identify time to metastatic disease in nmCRPC patients. Results: 654,148 patients were diagnosed with PC and received care in the VA from 2006 to 2021; of these, 12,992 developed nmCRPC, of whom 7,323 (56%) were white, 3,671 (28%) were black, 826 (6%) were Hispanic, and 1,172 (9%) were reported as other. Compared with white men at nmCRPC diagnosis, black men were younger (70 years vs. 74 years) while Hispanic men were older (75 years). White and Hispanic men had similar PSA at nmCRPC (10.2 ng/mL and 10.4 ng/mL), while the PSA for black men was higher (14.3 ng/mL). Similar treatment patterns were observed in all patients regardless of race; with ADT (60%), bicalutamide (24%), and finasteride (3%), the most frequently used first-line treatments. The mean time from nmCRPC to metastatic CRPC was 27.5 days for white men, 31.6 days for black men, 27.6 for Hispanic men, and 26.0 days for others. After adjusting for age, comorbidities, PSA doubling time, and treatment, the hazard ratio (HR) for developing metastatic disease was 0.85 (95% CI: 0.80–0.90, <0.001) for black men, 0.70 (95% CI: 0.62–0.78, <0.001) for Hispanic men, and 1.07 (95% CI: 0.99–1.16, <0.001) for others. Conclusions: These results suggest that when provided with care and treatment in an equal access healthcare system, black and Hispanic patients with nmCRPC are less likely to progress to metastatic CRPC than non-Hispanic white men.

The role of proteins implicated in skin steroidogenesis in acne vulgaris development

Background. Acne vulgaris belongs to androgen-dependent diseases where androgens cause sebaceous gland hypertrophy with a subsequent enlargement of sebum production.
 Aims. Determination of the expression profile of steroidogenesis-related proteins in normal skin and skin of patients with acne.
 Material and methods. Immunohistochemical assay was carried out to characterize TSPO, StAR, CYP11A1, and 3-hydroxysteroid dehydrogenase expression in skin of patients with acne vulgaris and in normal skin.
 Results. Expression levels of a protein TSPO which is involved in a steroidogenesis-limited step was increased in mature sebocytes of acne vulgaris patients. StAR, CYP11A1, and 3-hydroxysteroid dehydrogenase immunovisualization was found both in normal skin and in the skin of acne patients.
 Conclusions. The revealed alterations in expression of steroidogenesis-related protein in acne skin may indicate not only the conversion of steroids produced by external steroidogenesis organs but elevated steroids local production as well. The fact may account both for sebaceous gland hyperplasia and inflammation sustenance in skin.

Age, weight and circulating concentrations of total testosterone are associated with the relative prostatic size in adult intact male dogs

Prostatic hyperplasia (PH) is an androgen-dependent condition associated with increased prostatic size that is common in intact dogs, and similar to the condition in men. In dogs, the increase in prostatic size is most prominent the first years, and after approximately four years (in beagles), a plateau is reached, and further growth is slower. Why the prostate continues to grow more in some individuals is not clear. Most testosterone in the circulation is bound to albumin or sex hormone binding globulin (SHBG) and only a minor part is unbound and biologically active. The binding to SHBG has higher affinity than that to albumin. In addition, SHBG has own biological functions, modifying testosterone action. The aim of the present study was to investigate if there is an association between relative prostatic size and the variables total testosterone concentration, SHBG concentration, an estimation of bioavailable testosterone: the ratio between testosterone and SHBG (free androgen index, FAI), estradiol concentration, the estradiol/testosterone ratio, dog age and dog weight. Hormone concentrations were measured in serum from 79 intact male dogs aged ≥ four years, weighing ≥ five kg. The size of the prostate was estimated using ultrasonography, and relative prostate size, Srel, was calculated as the estimated size related to the normal size for a 4-year-old dog of the same weight. There as a negative correlation between testosterone concentration and age (ρ = −0.27, P = 0.018) and a positive correlation between age and Srel (ρ = 0.27, P = 0.016) and between SHBG and weight (ρ = 0.38, P = 0.001). The FAI was negatively correlated with dog weight (ρ = −0.32, P = 0.004). There were no significant correlations between Srel and SHBG or FAI or between estradiol or estradiol/testosterone and Srel, age or weight. A multiple regression analysis showed significant associations between log Srel and log testosterone concentration, log age and log weight of the dog, with an adjusted R2 of 9.5%. Although the variables total testosterone concentration, age and weight of the dog were all significantly associated with Srel, the coefficient of determination was low, indicating that they only explained a minor part of the prostatic size. The results support the analysis of total testosterone in studies of prostatic growth in the dog.

Cardiovascular complications of treatment for prostate cancer.

Prostate cancer, an androgen-dependent disease, is one of the leading causes of mortality in men. It can present as localised disease, locally advanced or distant metastatic disease. Treatment options for patients with prostate cancer include surgery, chemotherapy, brachytherapy, radiation therapy and hormonal therapy. There are multiple treatment options for each stage of the disease, but hormone therapy is usually reserved for advanced stages. Cardiovascular disease is the leading cause of death in patients with prostate cancer and both diseases share common risk factors. Hormone therapy improves prognosis in patients with more advanced disease, albeit at the cost of cardiovascular toxicity. Hormone therapy can be achieved with the use of agonists and antagonists of gonadotropin-releasing hormone receptors, androgen receptor blockers and enzyme inhibitors of androgen synthesis. Drug-specific cardiotoxicity caused by treatments for prostate cancer has not been fully elucidated. Cardiovascular disease in patients with prostate cancer is mainly managed via an ABCDE approach, a strategy to optimise common risk factors. With newer agents improving the prognosis for patients with prostate cancer, cardiovascular toxicity will have a greater impact on the outcomes of these patients. This article reviews cardiovascular risks associated with therapy for prostate cancer with a focus on hormonal therapy.

Synthesis, Characterization and Evaluation of 5α, 6β-Dihalo Androsterone Derivatives as 5α-Reductase Inhibitors

Background and Objective: Testosterone under the influence of 5α-reductase enzyme gets converted to dihydrotestosterone and high levels are found to be causative for androgen dependent disease like benign prostatic hyperplasia. Thus, 5α-reductase has been recognised as an important target for discovering new drugs against Benign Prostatic Hyperplasia and Prostate Cancer. Methods: In the present study, a series of 5α, 6β-Dichloro-17-Oxoandrostan-3β-yl esters (7a-7f) were synthesized and characterized by analytical and spectroscopic methods. The compounds were evaluated for their 5α-reductase inhibitory activity in-vivo by their effect on serum androgen level. Results: The target compounds (7a-7f) showed increased anti-androgenic activity as compared to finasteride and control, which implies that the target compounds are effective in inhibiting 5α-reductase. Particularly, compound 7b showing highest inhibitory activity and noteworthy D-Score was further sorted by performing solubility and dissolution studies. Results of these studies when compared with finasteride showed increased solubility and dissolution of target compound 7b. Conclusion: These results demonstrated that enhancement of activity by the presence of electronegative group at position 3 of the steroidal nucleus makes 7b a lead compound for further exploration and optimal formulation.

1698P Correlation between sex steroid hormone receptors expression in prostate cancer (PCa)

PCa is well known to be an androgen-dependent disease. However, estrogens and estrogen receptors alpha and beta (ERα and ERβ) also play an important role in PCa progression. All types of receptors share similar structure and can be activated by same ligands. As so, relative amounts of all receptor types may be important in predicting overall response to therapy, including androgen deprivation. We aimed to assess possible correlations between ERα, ERβ and androgen receptor (AR) expression in clinical PCa samples.

Developing New Treatment Options for Castration-Resistant Prostate Cancer and Recurrent Disease.

Prostate cancer (PCa) is a major diagnosed cancer among men globally, and about 20% of patients develop metastatic prostate cancer (mPCa) in the initial diagnosis. PCa is a typical androgen-dependent disease; thus, hormonal therapy is commonly used as a standard care for mPCa by inhibiting androgen receptor (AR) activities, or androgen metabolism. Inevitably, almost all PCa will acquire resistance and become castration-resistant PCa (CRPC) that is associated with AR gene mutations or amplification, the presence of AR variants, loss of AR expression toward neuroendocrine phenotype, or other hormonal receptors. Treating CRPC poses a great challenge to clinicians. Research efforts in the last decade have come up with several new anti-androgen agents to prolong overall survival of CRPC patients. In addition, many potential targeting agents have been at the stage of being able to translate many preclinical discoveries into clinical practices. At this juncture, it is important to highlight the emerging strategies including small-molecule inhibitors to AR variants, DNA repair enzymes, cell survival pathway, neuroendocrine differentiation pathway, radiotherapy, CRPC-specific theranostics and immune therapy that are underway or have recently been completed.

Abstract 1530: Derivatives of betulinic acid act as modulators of the androgen receptor and report cytotoxicity towards cancer cell lines

Abstract INTRODUCTION Triterpenoids are naturally occurring substances with a broad spectrum of beneficial effects for human health. Betulinic acid belongs to the group of pentacyclic triterpenoids and is well known for its antitumor activities. The library of about 1000 betulinic acid-derived triterpenes was collected and analyzed for cytotoxic activity on cancer and non-cancer cell lines (A549, CCRF-CEM, CEM-DNR, HCT116, K562, K562-TAX, U2OS, BJ and MRC-5) in our Institute over the last 20 years. Steroidal triterpenes are precursors of cholesterol, the precursor of testosterone, the fundamental ligand of the androgen receptor that is the key regulator in several physiological and pathological processes. Finding new modulators of androgen receptor is critical for the treatment of various androgen-dependent diseases, including prostate cancer or anabolic deficiencies. Betulinic acid and its derivatives showed structural similarity with steroids and therefore were selected to be tested for androgen receptor activity modulation. METHODS The stably transfected cell line HEK 293T GFP-AR was used to determine the translocation activity of the androgen receptor from the cytoplasm to the nucleus after the treatment with the derivatives. The high throughput fluorescent microscopy device and image analysis comprised the data, and compounds with the proved activity were selected for subsequent analysis. The luciferase reporter gene assay was further used to evaluate the effect of compounds on the transcriptional activity of the androgen receptor. The MTS assay was performed to assess the cytotoxicity. RESULTS Betulinic acid and its eight derivatives were identified to have 5-6 - fold higher translocation activity of AR (ratio cytoplasm GFP intensity/nuclear GFP intensity) than the negative control. Validation on luminescence luciferase reporter assay confirmed 4-5 - fold induction for five derivatives, whereas three derivatives were not active. Interestingly only two of them showed substantial cytotoxicities (IC50< 50 µM) as they possess significant inhibitory activity against all tested cancer cell lines with favorable therapeutic index against the non-cancer cell lines. CONCLUSION Derivatives of betulinic acid were proven to have modulatory activity on the androgen receptor pathway. The two derivatives of betulinic acid also displayed cytotoxicity towards almost all of the tested cancer cell lines. There could be not only AR modulation responsible for anticancer activity as the derivatives of betulinic acid are well know disruptors of mitochondrion functions as well. The synergy of these two mechanisms of action could potentiate the antitumor activity of the selected compounds. This work was supported by ENOCH CZ.02.1.01/0.0/0.0/16_019/0000868, CZ-OPENSCREEN - LM2018130, EATRIS-CZ - LM2018133. Citation Format: Alzbeta Srovnalova, Sona Gurska, Milan Urban, Jan Sarek, Jiri Rehulka, Petr Dzubak, Marian Hajduch. Derivatives of betulinic acid act as modulators of the androgen receptor and report cytotoxicity towards cancer cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1530.

Abstract SY33-02: The circadian cryptochrome, CRY1, is a pro-tumorigenic factor that rhythmically modulates DNA repair

Abstract The second leading cause of cancer-related death in US men is prostate cancer (PCa), which is an androgen-dependent disease in which the androgen receptor (AR) plays an important role. Recent studies have underscored the importance of AR-mediated DNA repair factor regulation as strongly associated with poor outcome in PCa, yet the mechanistic underpinnings of the molecular and cellular consequence of AR-regulated DNA repair and tumor progression have yet to be fully defined. Mining of comprehensive studies designed to identify AR functions critical for PCa revealed that AR binds to multiple regulatory elements within the CRY1 (cryptochrome 1) locus, as observed in several hormone therapy-sensitive PCa models.Here, the circadian factor CRY1, an evolutionally conserved transcriptional coregulator, is identified as a tumor specific regulator of DNA repair. Intriguingly, the N-terminus of CRY1 is a photolyase homology domain suggesting a potential role for CRY1 in DNA damage response. Circadian factors have been implicated in diverse non-canonical physiological functions such as DNA repair, DNA damage checkpoints, and apoptosis. Given the potential implications of CRY1 and DNA repair interplay, this study sought to delineate the molecular and biological consequences of androgen-responsive CRY1 in cancer through cistromic, transcriptomic, and molecular modeling of CRY1 dysregulation in PCa. Key findings demonstrate that CRY1 expression is androgen-responsive and associates with poor outcome in prostate cancer. Functional studies and first-in-field mapping of the CRY1 cistrome and transcriptome reveal that CRY1 regulates DNA repair and the G2/M transition. DNA damage stabilizes CRY1 in cancer (in vitro, in vivo, and human tumors ex vivo), which proves critical for efficient DNA repair. Further mechanistic investigation shows that stabilized CRY1 temporally regulates expression of genes required for homologous recombination. Collectively, these findings reveal that CRY1 is hormone-induced in tumors, is further stabilized by genomic insult, and promotes DNA repair and cell survival through temporal transcriptional regulation. These studies identify the circadian factor CRY1 as pro-tumorigenic and nominate CRY1 as a new therapeutic target. This work was recently published in Nature Communications (Shafi et al, 2021).Ongoing and future work with elucidate the role of CRY1 in PCa by identifying CRY1-driven pathways impacting DNA damage repair (DDR) regulation and CRY1 function on tumor progression and therapeutic response to discern novel pathways to improve existing therapies. Our ongoing work has the potential for broad implications for understanding and treating lethal PCa. Based on our preliminary data, we hypothesize that CRY1 is key modulator of DDR impacting PCa growth and survival, and that delineating the underlying molecular mechanisms of CRY1 and DDR interplay will allow for improved management of advanced disease. Our data indicate that CRY1 expression is altered in PCa, associated with poor outcome, and elicits pro-tumorigenic phenotypes through direct DDR regulation. Our continued work will define the biology of lethal PCa by delineating the molecular and cellular consequence of tumor-associated CRY1 deregulation, providing insight into the function of CRY1 on DNA repair competency. Also, this study will uncover mechanisms of action and identify novel partners for CRY1 function to transcriptionally regulate DNA repair and cancer progression. Lastly, the translational studies from this research study will elucidate the impact of CRY1 function on tumor progression and therapeutic response. Thus, these findings will reveal potential molecular and clinical biomarkers for patients and will ultimately aid in improving existing therapies and developing effective treatments to reduce death from PCa. The long-term impact of CRY1 as a pro-tumorigenic factor driving PCa progression and lethality, emphasizes the importance of developing treatments targeting this factor to mitigate aggressive disease. This study will be crucial in providing novel insight into CRY1 function in human PCa tumors to identify new avenues for therapeutic intervention. This research represents critical steps to understand how circadian factors, specifically CRY1, mediate DNA repair and promote tumorigenesis. Through our collaboration with circadian and DDR experts, this impactful work will not only provide deep mechanistic insight into CRY1 function in PCa, but also lay the foundation for preclinical studies aimed at developing novel strategies for treating CRY1-deregulated cancers. Thus, this ongoing research has immense translational impact serving as the basis for new investigator-initiated clinical trials and altering regimens to emphasize the importance of chronotherapy enhancing standard-of-care to ultimately improve outcome and reduce lethality of PCa. Citation Format: Ayesha Shafi. The circadian cryptochrome, CRY1, is a pro-tumorigenic factor that rhythmically modulates DNA repair [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr SY33-02.

Acne Vulgaris and Metabolic Syndrome: A Possible Association

IntroductionAcne vulgaris is an androgen-dependent disorder with excessive sebum production and proliferation of Propionibacterium acnes. Metabolic syndrome (MetS) is a multisystem disorder that increases the risk of diabetes mellitus, stroke, and cardiovascular diseases. ​This study aims to analyze the association of MetS with acne vulgaris.MethodsA cross-sectional study was conducted with 65 cases of acne vulgaris and 65 age and sex-matched controls. We used the system provided by the Indian authors for grading acne according to the clinical severity. In addition, the criteria updated according to the joint consensus of 2009 were employed for the diagnosis of MetS.ResultsOn clinical examination, grade 2 was the most prevalent grade of acne. We observed an increased incidence of abnormal waist circumference, triglyceride, HDL, and fasting blood glucose among the cases (p<0.05). Consequently, an increased occurrence of MetS was observed in the case group (p=0.011). While comparing the mean values of the parameters, we noted a significant difference in terms of waist circumference and HDL values. An increased mean value of waist circumference was noted in the case group while an increased mean value of HDL was reported from the control group (p<0.05). ConclusionPatients with acne vulgaris have a greater chance of developing MetS. Hence, an in-depth examination of clinical, anthropometric, and biochemical parameters that may lead to the development of MetS is necessary.