Clinical implications of single cell sequencing for bladder cancer.

Bladder cancer (BC) is the 10th most common cancer worldwide, with about 0.5 million reported new cases and about 0.2 million deaths per year. In this scoping review, we summarize the current evidence regarding the clinical implications of single-cell sequencing for bladder cancer based on PRISMA guidelines. We searched PubMed, CENTRAL, Embase, and supplemented with manual searches through the Scopus, and Web of Science for published studies until February 2023. We included original studies that used at least one single-cell technology to study bladder cancer. Forty-one publications were included in the review. Twenty-nine studies showed that this technology can identify cell subtypes in the tumor microenvironment that may predict prognosis or response to immune checkpoint inhibition therapy. Two studies were able to diagnose BC by identifying neoplastic cells through single-cell sequencing urine samples. The remaining studies were mainly a preclinical exploration of tumor microenvironment at single cell level. Single-cell sequencing technology can discriminate heterogeneity in bladder tumor cells and determine the key molecular properties that can lead to the discovery of novel perspectives on cancer management. This nascent tool can advance the early diagnosis, prognosis judgment, and targeted therapy of bladder cancer.

Healthcare Workers (HCWs) and non-HCWs reaction to Bacillus Calmette-Guérin (BCG) in the BATTLE trial.

OBJECTIVES: Healthcare workers (HCWs) may have different response to Bacillus Calmette-Guérin (BCG) vaccination due to previous occupational exposure to Mycobacterium particles. We report subgroup analysis of the BATTLE trial, comparing BCG effects in HCWs vs non-HCWs. This was a secondary analysis of a trial. METHODS: The BATTLE trial was a double-blind placebo-controlled phase III clinical trial that investigated BCG revaccinating adults who were recently infected with SARS-CoV-2 virus. BCG and placebo recipients were sub-grouped based on regular occupational contact with patients into HCWs (48 BCG and 50 placebo) and non-HCWs (124 BCG and 134 placebo). Weekly COVID-19 symptom progression and injection site reactions were compared between subgroups on weeks one, two, three, and six follow-ups. RESULTS: HCWs were more likely to complain of itching on the injection site early after injection (OR = 2.5, p = 0.049). They developed peeling and crusting on the site of injection faster than non-HCWs (during the second week, p = 0.033 and 0.040, OR = 3.3 and 2.7, respectively). HCWs were also more likely to maintain their papule or develop a late onset pustule during later weeks (weeks four and six, p = 0.024 and 0.006, OR = 2.2 and 8.6, respectively). In terms of COVID-19 symptom progression, recovery from anosmia was more likely in the non-HCWs who received BCG (week six, pHolm's corrected = 0.002, OR = 3.3). CONCLUSION: HCWs' local reaction to BCG injection was slightly more rapid and more intense, possibly due to their occupational exposure. BCG may also ameliorate COVID-19 induced inflammation and anosmia in non-HCWs but not HCWs. Therefore, HCWs might be less likely to benefit from BCG vaccination. CLINICALTRIALS: gov register number NCT04369794.

Bladder cancer immunomodulatory effects of intravesical Nitazoxanide, Rapamycin, Thalidomide and Bacillus Calmette-Guérin (BCG).

PURPOSE: To understand the effect of Nitazoxanide (NTZ), Rapamycin, Thalidomide, alone and in combination with BCG on bladder cancer (BC) histopathology and programmed death-ligand 1 (PD-L1) and anti-cytotoxic T lymphocyte antigen 4 (CTLA4) expression. METHODS: Female Fisher-344 rats underwent intravesical N-methyl-N-nitrosourea (MNU) followed by weekly intravesical treatment with saline (controls, n = 10), BCG (n = 10), NTZ (n = 8), BCG plus NTZ (n = 8), Rapamycin (n = 10) BCG plus Rapamycin (n = 10), Thalidomide (n = 10), and BCG plus Thalidomide (n = 10), and euthanized after 8 weeks and their bladders were investigated for BC and PD-L1 and CTLA4 expression. RESULTS: Rapamicyn alone and in combination with BCG had the lowest number of bladder neoplasias in the histopathology exam (1/10). Neoplastic lesions were found in 4/10 BCG recipients, 5/10 Thalidomide recipients, 4/10 Thalidomide plus BCG recipients, 5/8 NTZ and 3/8 NTZ plus BCG recipients. Adding NTZ to BCG increased the expression of PD-L1 and adding Rapamycin or Thalidomide decreased PD-L1 and CTLA4 expression compared to BCG alone. Rapamycin alone significantly increased CTLA4 and slightly increased PD-L1 expression but its combination with BCG significantly decreased both markers. Thalidomide had a similar effect; however, it was only slightly different from the control and BCG alone groups. CONCLUSION: Intravesical BCG combination treatment seems to effectively prevent BC development in an immunecompetent clinically relevant animal model, introducing Thalidomide, Nitazoxanide, and specially Rapamycin as candidates in the intravesical immunotherapy advancement. Our study contributes in understanding the mechanism of cancer immunotherapy.

Immune Checkpoint Glycoproteins Have Polymorphism: Are Monoclonal Antibodies Too Specific?

Since the 2018 Nobel prize in medicine was granted to the discovery of immune escape by cancer cells, billions of dollars have been spent on a new form of cancer immunotherapy called immune checkpoint inhibition (ICI). In this treatment modality, monoclonal antibodies (mAbs) are used to block cell-surface glycoproteins responsible for cancer immune escape. However, only a subset of patients benefit from this treatment. In this commentary, we focus on the polymorphism in the target molecules of these mAbs, namely PD-1, PD-L1 and CTLA4; we explain that using a single mAb from one clone is unlikely to succeed in treating all humans because humans have a genotype and phenotype polymorphism in these molecules. Monoclonal antibodies are highly specific and are capable of recognizing only one epitope ("monospecific"), which makes them ideal for use in laboratory animals because these animals are generationally inbred and genetically identical (isogenic). In humans, however, the encoding genes for PD-1, PD-L1 and CTLA4 have variations (alleles), and the final protein products have phenotype polymorphism. This means that small differences exist in these proteins among individual humans, rendering one mAb too specific to cover all patients. Our suggestion for the next step in advancing this oncotherapy is to focus on methods to tailor the mAb treatment individually for each patient or replace a single clone of mAb with less specific alternatives, e.g., a "cocktail of mAbs", oligoclonal antibodies or recombinant polyclonal antibodies. Fortunately, there are ongoing clinical trials on oligoclonal antibodies at the moment.

Comparing Bacillus Calmette-Guérin (BCG) strains in convalescent COVID-19 patients.

Aim: We previously published results of the BATTLE trial, showing that patients recently infected with SARS-CoV-2 can benefit from receiving Bacillus Calmette-Guérin (BCG) with minimal adverse effects. The study incorporated two strains of this vaccine. In this study, patient outcomes were compared based on the strain of BCG because different strains have been shown to have different immunogenicity. Methods: BATTLE was a double-blind controlled trial of COVID-19 convalescent patients; symptom progression, injection-site lesion characteristics and adverse effects were compared between recipients of placebo, Russian BCG strain or Brazilian BCG strains. Results: There was no statistically significant difference between the two BCG strains in terms of symptom progression, lesion-size or type. Conclusion: The two strains have similar clinical outcomes in COVID-19 convalescent patients.

We previously published results of the BATTLE trial, showing that patients recently infected with SARS-CoV-2 virus can benefit from receiving BCG with minimal adverse effects. This article shows that the two BCG strains, Russian and Brazilian, have similar clinical outcomes in COVID-19 convalescent patients.

Time Analysis of an Emergent Infection Spread Among Healthcare Workers: Lessons Learned from Early Wave of SARS-CoV-2.

Aim: To find whether an emergent airborne infection is more likely to spread among healthcare workers (HCW) based on data of SARS-CoV-2 and whether the number of new cases of such airborne viral disease can be predicted using a method traditionally used in weather forecasting called Autoregressive Fractionally Integrated Moving Average (ARFIMA). Methods: We analyzed SARS-CoV-2 spread among HCWs based on outpatient nasopharyngeal swabs for real-time polymerase chain reaction (RT-PCR) tests and compared it to non-HCW in the first and the second wave of the pandemic. We also generated an ARFIMA model based on weekly case numbers from February 2020 to April 2021 and tested it on data from May to July 2021. Results: Our analysis of 8998 tests in the 15 months period showed a rapid rise in positive RT-PCR tests among HCWs during the first wave of pandemic. In the second wave, however, positive patients were more commonly non-HCWs. The ARFIMA model showed a long-memory pattern for SARS-CoV-2 (seven months) and predicted future new cases with an average error of ±1.9 cases per week. Conclusion: Our data indicate that the virus rapidly spread among HCWs during the first wave of the pandemic. Review of published literature showed that this was the case in multiple other areas as well. We therefore suggest strict policies early in the emergence of a new infection to protect HCWs and prevent spreading to the general public. The ARFIMA model can be a valuable forecasting tool to predict the number of new cases in advance and assist in efficient planning.

Immunogenic Cell Death Role in Urothelial Cancer Therapy.

PURPOSE: Bladder cancer is the 13th most common cause of cancer death with the highest lifetime cost for treatment of all cancers. This scoping review clarifies the available evidence on the role of a novel therapeutic approach called immunogenic cell death (ICD) in urothelial cancer of the bladder. METHODS: In accordance with the recommendations of the Joanna Briggs Institute, we searched MEDLINE (Ovid), EMBASE, CENTRAL databases, and supplemented with manual searches through the conferences, Google scholar, and clinicaltrials.gov for published studies up to April 2022. We included literature that studied molecular mechanisms of ICD and the role of certain danger-associated molecular patterns (DAMPs) in generating ICD, safety and efficacy of different ICD inducers, and their contributions in combination with other urothelial cancer treatments. RESULTS: Oncolytic viruses, radiotherapy, certain chemo/chemo radiation therapy combinations, photodynamic therapy, and novel agents were studied as ICD-inducing treatment modalities in the included studies. ICD was observed in vitro (murine or human urothelial carcinoma) in ten studies, eight studies were performed on mouse models (orthotopic or subcutaneous), and five clinical trials assessed patient response to ICD inducing agents. The most common studied DAMPs were Calreticulin, HMGB1, ATP, and Heat Shock Proteins (HSP) 70 and 90, which were either expressed on the cancer cells or released. CONCLUSION: ICD inducers were able to generate lasting antitumor immune responses with memory formation in animal studies (vaccination effect). In clinical trials these agents generally had low side effects, except for one trial, and could be used alone or in combination with other cancer treatment strategies in urothelial cancer patients.

Digital rectal examination impact on PSA derivatives and prostate biopsy triggers: a contemporary study.

OBJECTIVE: To evaluate the impact of the digital rectal exam (DRE) on PSA measurements and clinical decision-making. METHODS: Healthy male volunteers between 50 and 70 years old were recruited during a 30-day public screening program. PSA levels were measured using two different methods (standard enhanced chemiluminescence immunoassay-ECLIA, and novel immunochromatography assay-ICA/rapid PSA) in the same blood sample. Two blood samples were drawn; first before DRE and the second 30-40 min after DRE. The effect of DRE on PSA levels and its impact on clinical decision-making for individual patients were evaluated based on different biopsy trigger cutoffs. RESULTS: ECLIA-PSA was measured in 74 participants both pre- and 37 ± 5 min post-DRE, mean age 57.2 ± 8.3 years, and mean prostate volume 33.6 (20-80) cm3. Both total and free ECLIA-PSA increased significantly after DRE (mean increase of 0.47 and 0.26 ng/ml, respectively, both p < 0.001). Different internationally accepted biopsy triggers were reached after DRE only: 5 total PSA > 3 ng/ml, 13 increase > 0.75 ng/ml, 3 PSA density > 0.15, and 1 free/total PSA < 0.18. On two occasions, patients were pushed away from biopsy trigger after DRE due to free/total PSA > 0.18. ICA-PSA was detectable (> 2.0 ng/ml) in 5 of 45 measured samples (11%) before DRE and 13/45 (29%) after DRE, p = 0.0316. Four among five detectable ICA-PSA tests increased after DRE. CONCLUSION: Performing DRE immediately before PSA measurement might change the clinical decision-making on a significant number of occasions (roughly 1 in 3); even though the mean increase (0.47 ng/ml) looks deceivingly small. Further studies are required that include gold standard tests (biopsy, or imaging).

BCG vaccine safety in COVID-19 convalescent adults: BATTLE a randomized controlled trial.

INTRODUCTION: The safety of BCG revaccination is uncertain and there is no data on its use in patients with COVID-19. METHODS: COVID-19 convalescent adults confirmed by SARS-CoV-2 RT-PCR in South-America were 1:1 randomized in the first 14 days of symptoms to BCG intradermal vaccine or placebo and evaluated for adverse events on days 7, 14, 21, and beyond 40 days. CLINICAL TRIAL REGISTRATION: NCT04369794. RESULTS: 151 placebo and 148 BCG patients were included in the final analysis, with an average age of 40.7 years. No severe adverse event to BCG was reported. On day 7, 130 (87.8%) of the BCG recipients had local reaction, average size of 10.6 ± 6.4 mm, compared to only 2 (1.3%) placebos. Lesions gradually shrunk in size (mean 10.5 mm, 9.7 mm, and 6.8 mm at 14, 21, and beyond 40 days, respectively. The number of symptoms in any of the visits was not different between groups, and anosmia resolved earlier (25.7% vs. 37.1% at 7 days, OR = 1.70, 1.01-2.89, p = 0.035) in the BCG recipients. CONCLUSION: The BCG revaccination is safe in convalescent COVID-19 adults of a tuberculosis endemic region, regardless of tuberculin or IGRA test results. Local adverse events were similar though occurred earlier to that previously reported in children.

Randomized clinical trial of BCG vaccine in patients with convalescent COVID-19: Clinical evolution, adverse events, and humoral immune response.

BACKGROUND: The Bacillus Calmette-Guérin (BCG) vaccine may confer cross-protection against viral diseases in adults. This study evaluated BCG vaccine cross-protection in adults with convalescent coronavirus disease 2019 (COVID-19). METHOD: This was a multicenter, prospective, randomized, placebo-controlled, double-blind phase III study (ClinicalTrials.gov: NCT04369794). SETTING: University Community Health Center and Municipal Outpatient Center in South America. PATIENTS: a total of 378 adult patients with convalescent COVID-19 were included. INTERVENTION: single intradermal BCG vaccine (n = 183) and placebo (n = 195). MEASUREMENTS: the primary outcome was clinical evolution. Other outcomes included adverse events and humoral immune responses for up to 6 months. RESULTS: A significantly higher proportion of BCG patients with anosmia and ageusia recovered at the 6-week follow-up visit than placebo (anosmia: 83.1% vs. 68.7% healed, p = 0.043, number needed to treat [NNT] = 6.9; ageusia: 81.2% vs. 63.4% healed, p = 0.032, NNT = 5.6). BCG also prevented the appearance of ageusia in the following weeks: seven in 113 (6.2%) BCG recipients versus 19 in 126 (15.1%) placebos, p = 0.036, NNT = 11.2. BCG did not induce any severe or systemic adverse effects. The most common and expected adverse effects were local vaccine lesions, erythema (n = 152; 86.4%), and papules (n = 111; 63.1%). Anti-severe acute respiratory syndrome coronavirus 2 humoral response measured by N protein immunoglobulin G titer and seroneutralization by interacting with the angiotensin-converting enzyme 2 receptor suggest that the serum of BCG-injected patients may neutralize the virus at lower specificity; however, the results were not statistically significant. CONCLUSION: BCG vaccine is safe and offers cross-protection against COVID-19 with potential humoral response modulation. LIMITATIONS: No severely ill patients were included.

Effect of BCG on Respiratory Complications Caused by COVID-19: A Scoping Review.

Introduction: Bacillus Calmette-Guérin (BCG) has been shown to have protective effects against respiratory viruses. We conducted a scoping review of the literature to clarify the available evidence regarding the effect of BCG therapy in preventing respiratory complications of coronavirus disease 2019 (COVID-19). Methods: We searched PubMed, Embase, CENTRAL, Scopus, and Web of Science for related studies up to October 2022. Results: In total, 35 publications and trials were included. One animal study, two observational studies, and six finalized trials measured the effect of BCG administration on respiratory complications of COVID-19. The remaining publications included eight unfinished trials, 12 ecological studies, and six observational studies that did not directly measure respiratory complications but assessed overall mortality of the disease and were included as an adjunct to our study. All trials involved vaccinating adults to protect them against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection, and measured respiratory symptoms or the need for intensive respiratory support as the primary or secondary aim of the study. One trial that exclusively included at-risk adults between 18 and 60 years old showed a decreased chance of respiratory complications as the secondary outcome of the study. Another trial that exclusively evaluated this effect on the elderly (60 years and older) as the primary aim of the study reported no protective effect against respiratory complications. The remaining literature provided mostly inconclusive evidence. Conclusion: The majority of the literature on the protective effect of BCG against respiratory complications of COVID-19 is inconclusive.

Androgen Receptor Signaling Reduces the Efficacy of Bacillus Calmette-Guérin Therapy for Bladder Cancer via Modulating Rab27b-Induced Exocytosis.

Although intravesical bacillus Calmette-Guérin (BCG) immunotherapy has been the gold standard for nonsurgical management of non-muscle-invasive bladder cancer, a considerable number of patients exhibit resistance to the adjuvant treatment with unexplained mechanisms. This study aimed to investigate whether and how androgen receptor (AR) signals modulate BCG cytotoxicity in bladder cancer. AR knockdown or overexpression in bladder cancer lines resulted in induction or reduction, respectively, in intracellular BCG quantity and its cytotoxic activity. Microarray screening identified Rab27b, a small GTPase known to mediate bacterial exocytosis, which was upregulated in BCG-resistant cells and downregulated in AR-shRNA cells. Knockdown of Rab27b, or its effector SYTL3, or overexpression of Rab27b also induced or reduced, respectively, BCG quantity and cytotoxicity. In addition, treatment with GW4869, which was previously shown to inhibit Rab27b-dependent secretion, induced them and reduced Rab27b expression in bladder cancer cells. Meanwhile, AR expression was upregulated in BCG-resistant lines, compared with respective controls. In a mouse orthotopic xenograft model, Rab27b/SYTL3 knockdown or GW4869 treatment enhanced the amount of BCG within tumors and its suppressive effect on tumor growth. Moreover, in non-muscle-invasive bladder cancer specimens from patients subsequently undergoing BCG therapy, positivity of AR/Rab27b expression was associated with significantly higher risks of tumor recurrence. AR activation thus correlates with resistance to BCG treatment, presumably via upregulating Rab27b expression. Mechanistically, it is suggested that BCG elimination from urothelial cells is induced by Rab27b/SYTL3-mediated exocytosis. Accordingly, Rab27b inactivation, potentially via antiandrogenic drugs and/or exocytosis inhibition are anticipated to sensitize the efficacy of BCG therapy, especially in patients with BCG-refractory AR/Rab27b-positive bladder cancer.

Twelve years' experience with fascia lata autograft to replace complicated anterior vaginal mesh.

OBJECTIVES: To report 12-year experience with replacing transvaginal mesh (TVM) with fascia lata autograft. METHODS: This was a chart review of TVM removal and replacement with a fascia lata autograft placement by a single surgeon between 2005 and 2017. The Pelvic Organ Prolapse Quantification (POP-Q) system before and 1 year following the procedure, patient-reported recurrence of symptoms, changes in the POP-Q examination and complication rates are analyzed. RESULTS: Twenty-four patients were included. Mean age was 57.2 (95% CI 53.2-61.2) years. Mean number of days to Foley catheter removal was 3.2 days (95% CI 1.6-4.9) and mean number of days to drain removal was 10.9 days (95% CI 9.9-12.0). Following the surgery, no leg seroma, infection or numbness was reported. UTI occurred in four (16.7%) of the participants postoperatively. At 3-month follow-up, mild urinary symptoms were reported in five participants (20.8%). At 1-year follow-up, one participant was symptomatic of pelvic organ prolapse. Paired t-test analysis revealed statistically significant retraction of Aa and Ba vaginal points (p < 0.001). C, GH and PB points were also statistically significantly retracted. CONCLUSION: Fascia lata autograft for anterior compartment reconstruction due to TVM complications is associated with high safety and efficacy rates.

Ultrasongraphic age-related changes of the pelvic floor muscles in nulliparous women and their association with pelvic floor symptoms: A pilot study.

OBJECTIVE: To assess age-related changes in the pelvic floor muscular hiatus and their association with symptoms of pelvic organ prolapse, urinary and fecal incontinence, and sexual function. METHODS: In this pilot study we performed 3D endovaginal ultrasonography in two age groups of nulliparous women: 18 to 40 years and 52 to 85 years. Anterior-posterior (AP) diameter, left-right (LR) diameter, and the Minimal Levator Hiatus area were measured. The AP/LR ratio was calculated to compare the shape of the pelvic floor muscles between participants (oval vs circular). Other measurements included length of the urethra, and levator plate lift. Participants were assessed for (1) distress symptoms of pelvic floor prolapse, urinary, and fecal symptoms by the Pelvic Floor Distress Inventory-20, (2) quality of life via the pelvic floor impact inventory-7, and (3) sexual function by the female sexual function inventory (FSFI-19). RESULTS: A total of 12 women into the younger group and 10 to the older group were recruited. Older women had higher AP/LR ratio and longer distance levator plate lift while performing the squeeze maneuver ( P = 0.017 and 0.038, respectively). Older women had worse urinary and pelvic organ prolapse symptoms ( P = 0.002 and 0.004, respectively). Fewer women in the older group were sexually active (60% vs 92%) and their quality of sexual life was lower based on their FSFI-19 results. CONCLUSION: Levator ani muscle hiatus changes to a more oval form in older nulliparous postmenopausal women and this change in shape is associated with increased pelvic floor symptoms.

Six-Year Experience in Teaching Pelvic Floor Ultrasonography Using Pelvic Floor Phantoms.

OBJECTIVE: To review our 6-year experience (2009-2015) in teaching three-dimensional pelvic floor ultrasonography workshops that utilized pelvic floor phantoms in the setting of an Objective Structured Assessment of Technical Skills methodology. METHODS: Four-hour Objective Structured Assessment of Technical Skills workshops were given at several society meetings and involved a didactic session, a hands-on session using the pelvic floor phantoms, and a computer station session reviewing pelvic floor pathologies. We analyzed improvement in participants' diagnostic skills using a test with 60 illustrated questions of normal and pathologic findings in live human models. RESULTS: Two hundred forty-three attendees completed the 60-question test before and after attending the workshop. Paired t test showed a significant improvement in attendees' average scores after the workshop in all categories: anatomy, normal, or pathologic endovaginal imaging and normal or pathologic endoanal imaging (P<.001 for all). McNemar test showed a statistically significant increased number of correct answers in 50 of 60 (83%) questions. CONCLUSION: Our Objective Structured Assessment of Technical Skills workshops incorporating pelvic floor phantoms enhanced trainees' pelvic floor ultrasound diagnostic skills.

Davydov Procedure for Augmenting Vaginal Length in a Postsurgical Male-to-Female Transgender Patient.

Male-to-female gender reassignment surgery may lead to unsatisfactory vaginal length. No standard approaches are established to treat these patients. We present a case of vaginoplasty using the laparoscopic Davydov technique performed on a male-to-female transgender patient. Our case provides a novel approach to treating this rare condition and introduces Davydov procedure as a potentially effective and safe treatment. Preoperative endovaginal and endoanal 3-dimensional ultrasounds and a pelvic magnetic resonance imaging of the case are presented to provide details of this patient's unique anatomy.

Compound A Inhibits Bladder Cancer Growth Predominantly via Glucocorticoid Receptor Transrepression.

Recent evidence indicates that glucocorticoids (GCs) suppress bladder cancer cell invasion through the GC receptor (GR) pathway, whereas androgen-mediated androgen receptor (AR) signals induce bladder tumor progression. In this study, we assessed the effects of 2-(4-acetoxyphenyl)-2-chloro-N-methyl-ethylammonium chloride (compound A [CpdA]), which was shown to function as not only a GR modulator but also an AR antagonist, on the growth of bladder cancer. In GR/AR-positive cells, CpdA strongly inhibited cell proliferation and colony formation as well as increased G1 phase-arrested cell population and apoptosis. Specifically, CpdA at 1µM decreased cell viability of TCCSUP/UMUC3-control-short hairpin RNA (shRNA), TCCSUP/UMUC3-GR-shRNA, and TCCSUP/UMUC3-AR-shRNA by 50%/67%, 25%/26%, and 38%/58%, respectively. CpdA also inhibited cell migration and invasion of GR/AR-positive (up to 61% decrease) and GR-positive/AR-silencing (up to 51% decrease) lines and, less strongly, those of GR-silencing/AR-positive lines (up to 35% decrease). Additionally, in UMUC3-control xenograft-bearing male mice, CpdA more strongly suppressed tumor growth than dexamethasone or hydroxyflutamide. In reporter gene assays, CpdA failed to induce GR transactivation, whereas it antagonized dihydrotestosterone-enhanced AR transactivation. In contrast, CpdA reduced nuclear factor (NF)-κB and activator protein 1 transcriptional activities, indicating induction of GR-mediated transrepression. Correspondingly, the expression of NF-κB-related molecules, matrix metalloproteinase-2, matrix metalloproteinase-9, interleukin-6, and vascular endothelial growth factor, was significantly down-regulated by CpdA in control lines but not in GR-silencing cells. Moreover, coimmunoprecipitation showed that CpdA promoted the interactions between GR and NF-κB. Thus, CpdA likely inhibits bladder cancer growth predominantly via inducing GR transrepression and at least partially mediated through the AR pathway, suggesting its effects more beneficial than GCs/pure GR ligands or AR antagonists.