Search Results (1,295)

Nectin-4 has emerged as a clinically relevant target in muscle-invasive bladder cancer (MIBC), primarily because of its role in antibody–drug conjugate-based therapies. However, the broader biological context of Nectin-4 expression and its association with tumor-promoting signaling pathways in MIBC remain insufficiently characterized. In this single-institution study, Nectin-4 expression (H-score 0–300) was assessed by immunohistochemistry in two independent MIBC cohorts. Associations between Nectin-4 expression and key markers related to growth signaling, metabolic regulation, and inflammation were analyzed alongside clinicopathological characteristics. Nectin-4 expression was significantly higher in malignant tissue than in non-malignant tissue (p = 0.0016 and p = 0.0302, respectively). Nectin-4 expression was not associated with demographic or clinicopathological parameters; however, a trend toward lower expression in more advanced disease stages was observed. Significant positive correlations were identified between Nectin-4 expression and protein kinase B (p = 0.0004), cytoplasmic (p = 0.0115) and membranous somatostatin receptor 2 (p = 0.0125), insulin receptor substrate 1 (p = 0.03), and interleukin-1 receptor antagonist (IL-1RA; p = 0.0045). In contrast, a negative correlation was observed with the IL-1β/IL-1RA ratio (p = 0.0246). Although Nectin-4 expression was not significantly associated with cancer-specific or overall survival, a trend toward shorter relapse-free survival was observed in patients with lower Nectin-4 expression (p = 0.0531). In multivariate analysis, patient age, but not Nectin-4 expression, emerged as an independent prognostic factor. Although Nectin-4 expression does not appear to have independent prognostic value, its biological associations suggest that it reflects an integrated tumor-related signaling context. These findings support further investigation of Nectin-4 as part of rational, biology-driven therapeutic strategies in bladder cancer. Full article

Background. Accurate prediction of pathologic complete response (pCR) to neoadjuvant chemotherapy (NAC) in muscle-invasive urothelial bladder cancer (MIBC) remains an unmet clinical need. The neutrophil-to-platelet/hemoglobin-to-lymphocyte (NPLH) ratio, a composite hematologic index that reflects both systemic inflammation and nutritional oxygen-carrying capacity, has not been previously evaluated as a predictor of NAC response in this setting. Methods. We retrospectively analyzed 114 consecutive patients with MIBC (cT2–T4, N0–N3) who received accelerated MVAC (aMVAC) NAC followed by radical cystectomy at a single academic center. Pretreatment NPLH (calculated as [neutrophils × platelets]/[hemoglobin × lymphocytes]) was assessed as a predictor of pCR (ypT0N0) and tumor regression grade (TRG). Receiver operating characteristic (ROC) curve analysis, Mann–Whitney U test, and logistic regression were used. NPLH performance was compared to NLR and PLR. Results. pCR was achieved in 35 patients (30.7%). Median NPLH was significantly lower in pCR vs. non-pCR patients (33.9 [IQR 23.1–42.4] vs. 47.6 [IQR 30.7–90.4]; p = 0.0007). NPLH yielded an AUC of 0.700 (bootstrap 95% CI 0.596–0.794) for pCR prediction, numerically superior to NLR (AUC 0.645 [0.542–0.741]) and PLR (AUC 0.643 [0.533–0.747]); DeLong test: NPLH vs. NLR p = 0.079, NPLH vs. PLR p = 0.090. At the optimal cut-off of 44.5, NPLH demonstrated 80.0% sensitivity and 57.0% specificity. pCR rates declined progressively across NPLH quartiles: 48.3% (Q1) to 10.3% (Q4). On multivariate logistic regression, log-transformed NPLH was the only independent predictor of pCR (parsimonious model, OR 0.292, 95% CI 0.131–0.652; p = 0.003; EPV = 17.5). A positive correlation was observed between NPLH and TRG score (Spearman r = 0.284; p = 0.0022), with significant differences between TRG 1 and TRG 3 subgroups (p = 0.0036). Conclusions. Pretreatment NPLH is an independent predictor of pCR to aMVAC in MIBC and is numerically superior to NLR and PLR (DeLong p = 0.079). Consisting exclusively of standard complete blood count parameters, NPLH is readily available and inexpensive. This single-center exploratory study is hypothesis-generating and requires prospective external validation before clinical implementation. Full article

Bladder cancer is a common malignancy with high recurrence rates and significant morbidity, necessitating accurate diagnostic and prognostic tools. Although cystoscopy and transurethral resection of bladder tumor (TURBT) remain reference standards, these approaches are invasive and limited by sampling errors and understaging. Consequently, there is growing interest in non-invasive biomarkers, including urine-based assays, blood-based markers, and imaging-derived parameters. Among these biomarkers, multiparametric magnetic resonance imaging (mpMRI), particularly with the Vesical Imaging Reporting and Data System (VI-RADS), has emerged as a robust non-invasive imaging biomarker for local staging and risk stratification. Recent evidence suggests that mpMRI plays a role in predicting treatment response and recurrence, particularly in the context of neoadjuvant therapy. This review provides a comprehensive overview of current non-invasive diagnostic and prognostic biomarkers in bladder cancer, with a particular emphasis on imaging biomarkers. We discuss their clinical utility, limitations, and future integration into multimodal decision-making frameworks. Full article

The failure of therapy in muscle invasive bladder cancer (MIBC) is primarily attributed to tumor heterogeneity and therapy resistance. We propose a novel approach targeting interleukin-13 receptor subunit alpha 2 (IL-13Rα2), which is expressed on bladder cancer (BC) cells but absent in normal urothelial cells. We investigated the therapeutic effects of WPD101a immunotoxin (IL-13-DT390) on IL-13Rα2-expressing BC cells in relation to BC cell phenotype and functional characteristics in vitro using both 2-dimensional (2D) and 3-dimensional (3D) models. Cell phenotype and IL-13Rα2 expression were assessed using flow cytometry, immunofluorescence, and Western blot analysis. The biological effects of WPD101a were evaluated by measuring cell viability and proliferation using the MTT, sulforhodamine B (SRB), CellTiter-Glo and Live/Dead assays. Apoptosis was assessed using Annexin V/propidium iodide (PI) staining, and quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis of CASP genes expression. We found that the reference BC cell lines TCC-SUP, JMSU-1 and UM-UC-3 express IL-13Rα2 at various level in contrast to RT-4, HCV-29 and 5637 cells. Cells expressing IL-13Rα2 were sensitive to WPD101a at lower concentrations in the 2D model (0.1 ng/mL) compared to the 3D model (1.0 ng/mL). IL-13Rα2-negative cells remain resistant to the immunotoxin. WPD101a induces apoptosis in BC cells expressing IL-13Rα2 as confirmed by the presence of apoptotic cells, increase the proportion of cells in the subG1 phase, and by the effector CASP3, CASP7, and initiator CASP8, CASP9 genes expression. This study confirmed receptor-dependent cytotoxic effects of WPD101a and the ability and specificity to inhibit growth and apoptosis induction in MIBC cells expressing IL-13Rα2. Full article

Background/Objectives: Lymph node metastasis within the prostatic anterior fat pad (PAFP) is uncommon but may refine nodal staging when pelvic lymph node dissection and PSMA PET/CT are negative. Case Presentation: A 58-year-old man with PSA 59 ng/mL, negative digital rectal examination, and a PI-RADS 5 anterior lesion underwent transperineal MRI/US fusion biopsy, showing an acinar adenocarcinoma (Gleason score 5 + 5 = 10, ISUP grade group 5) confined to anterior cores. 18F-PSMA-1007 PET/CT showed intense intraprostatic uptake (SUVmax 55.2) without nodal or distant disease. Retropubic radical prostatectomy, bilateral extended pelvic lymph node dissection (ePLND), and separate PAFP submission were performed. Final pathology showed a 38 mm bilateral anterior tumor involving 35% of the prostate, focal anterior extraprostatic extension, negative margins, absent seminal vesicle and bladder neck invasion, perineural and lymphovascular invasion, and no cribriform or intraductal carcinoma. All 15 pelvic nodes were negative. One of two PAFP nodes contained a 3 mm PSA-positive metastasis without extranodal extension, resulting in pT3aN1 staging. Postoperative PSA persistence prompted radiotherapy plus androgen deprivation therapy; PSA was 0.01 ng/mL at 6 months. Conclusions: In very-high-risk anterior prostate cancer, separate PAFP evaluation may provide clinically relevant staging information when PSMA PET/CT and pelvic lymph nodes are negative. This case highlights the PAFP as a potential site of occult regional nodal disease. Full article

Endocrine-disrupting chemicals (EDCs) are important environmental risk factors for urogenital malignancies, but the shared molecular mechanisms underlying their carcinogenic effects remain poorly understood. Here, we systematically investigated the common pro-tumorigenic mechanisms of 12 prevalent EDCs, including anthracene, benzo[a]pyrene (BaP), bisphenol A, clofenotane, di(2-ethylhexyl) phthalate, diazinon, dibutyl phthalate, glyphosate, malathion, perfluorooctanoic acid, polychlorinated biphenyls, and triclosan, across four urogenital cancers, including bladder cancer (BLCA), renal cell carcinoma (RCC), prostate adenocarcinoma (PRAD), and testicular germ cell tumor (TGCT). By integrating network toxicology and protein–protein interaction analysis, we identified shared hub targets linking EDC exposure to tumor progression. EGFR and CASP3 were identified as core targets in BLCA, EGFR and CASP9 in RCC, and CASP3, ESR1, and EGFR in PRAD, whereas KIT emerged as a broadly relevant target in TGCT. Molecular docking and molecular dynamics simulations supported the stable binding of EDCs to these targets. Among the predicted interactions, BaP showed strong binding affinity for CASP9 (ΔG = −9.8 kcal/mol) and was therefore selected for experimental validation. Analysis of TCGA data showed that elevated CASP9 expression was significantly associated with poorer overall survival in patients with RCC. In 786-O and ACHN cells, chronic exposure to an environmentally relevant concentration of BaP significantly increased CASP9 protein stability without altering its mRNA expression, suggesting post-transcriptional regulation. Collectively, these findings identify shared molecular targets of EDCs across urogenital cancers and provide new mechanistic insight into EDC-driven tumor progression, prioritizing potential biomarkers and therapeutic targets for environmentally related malignancies. Full article

Background and Objectives: Bladder cancer (BCa) is characterized by high rates of recurrence and progression, underscoring the need for reliable non-invasive biomarkers. Circular RNAs (circRNAs) are covalently closed non-coding RNAs generated by back-splicing and are stable in biological fluids, including urine. Increasing evidence implicates circRNAs in BCa pathogenesis. However, identification of clinically relevant circRNAs remains challenging. This study aimed to streamline circRNA selection and identification of functional urinary circRNAs for potential use as biomarkers for BCa monitoring. Methods: Using a database-screening approach, we identified circRNAs with high predicted affinity (TDMD score > 1.1) to miR-101-3p (a tumor-suppressive microRNA in BCa). In addition, candidate circRNAs were prioritized based on the following: (i) derivation from genes involved in BCa tumorigenesis; and (ii) origination from exonic or long non-coding RNA sequences. The potential contribution of Argonaute-2 (Ago2) binding sites to circRNA activity or potential usage as biomarker was also evaluated. Expression levels were assessed in urine samples and BCa cell lines, and functional relevance was examined using molecular and cellular assays. Results: circCIAO1(5) and circMALAT1 fulfilled prioritization criteria and exhibited distinct Ago2-binding site profiles. Both circRNAs were upregulated in urine from BCa patients and in aggressive BCa cell lines and showed differential expression between remission and recurrent disease. CircCIAO1(5) demonstrated higher-affinity binding to miR-101-3p, while both circRNAs interacted with miR-101-3p and Ago2. Functional assays revealed enhanced proliferation, motility, and invasion upon circRNA overexpression, consistent with miR-101-3p sequestration and reduction in depression of its target oncogene—EZH2. Conclusions: circCIAO1(5) and circMALAT1 are promising candidates as urinary biomarkers for noninvasive BCa monitoring, illustrating the value of bioinformatics-guided determination of circRNA as potential biomarkers and significance of circRNA-mediated regulatory mechanisms in BCa biology. Full article

Background: Most bladder cancer cases present as non-muscle-invasive bladder cancer (NMIBC), with the course of multiple recurrences leading to stage progression to muscle-invasive bladder cancer (MIBC) in 10–20% of cases, which is associated with higher morbidity and mortality. Accurate histopathologic classification of bladder cancer remains important for patient management. Methods: This retrospective–prospective observational cohort study was conducted on 244 transurethral resection specimens. Immunohistochemistry assessed CK5/6, CK20, and apoptosis-inducing factor (AIF) using three representations: intensity, percentage of positive cells, and multiplicative score. Discrimination between NMIBC (pTa/pT1) and MIBC (≥pT2), and between low-grade (LG) and high-grade (HG) tumors, was evaluated using ROC/AUC analysis and logistic regression. The main analysis focused on cross-sectional marker performance in primary/non-recurrent tumors. Recurrent tumors were analyzed only as an exploratory subgroup. Tumors were also categorized into basal, luminal, mixed/double-positive, and double-negative phenotypes using thresholds of 10% for CK5/6 and CK20. Results: For stage discrimination, all three markers showed modest separation. The best-performing representation was CK5/6 intensity (AUC 0.641; lower in MIBC). For grade discrimination, the AIF score showed the highest performance (AUC 0.729, higher in HG). Combining markers improved model performance (NMIBC vs. MIBC: AUC 0.784; strict LG vs. HG: AUC 0.778). Using the 10% cutoff in non-recurrent tumors, mixed/double-positive tumors had the lowest MIBC proportion (6.0%) and double-negative tumors the highest (46.7%). Conclusions: CK5/6, CK20, and AIF provide modest discrimination between stages, with lower CK5/6 and CK20, and higher AIF, in MIBC. The AIF score shows the highest separation between grades and may serve as a useful non-proliferation marker for grading, particularly when interpreted alongside CK5/6 and CK20 in a simple immunohistochemical panel. Full article

Background/Objectives: Urothelial carcinomas are the most common tumors of the bladder. There are limited known cancer stem cell markers in these tumors. Ly6/uPAR gene family members are considered to be markers of cancer stem cells and tissue stem cells in mice, but studies on their expression or role in human cancers are limited. In this study, we aimed to investigate the expression of LY6/uPAR gene family members in human urothelial cancers. Methods: A total of 84 patients were included in the study. Patients diagnosed with urothelial carcinoma were divided into low-grade noninvasive and high-grade invasive carcinoma groups. Normal urothelial samples were used as a control group. RNA isolation was performed from paraffin blocks, and then cDNA was obtained. LY6D, LY6E, LY6H, LY6K, PSCA, LYPD2, SLURP1, GML, GPIHBP1, and LYNX1 genes were analyzed by qRT-PCR method. Results: We observed significantly higher expression of LY6E, LY6K, PSCA, GPIHBP1, and LYNX1 genes in urothelial carcinomas, but lower expression of LY6H, LYPD2, and SLURP1 genes in urothelial cancers compared to the control tissue. Decreased expression of LY6H, PSCA, LYPD2, SLURP1, and GPIHBP1 genes was significantly correlated with poor survival. Conclusions: In the present study, the expression of this gene family in bladder cancer was investigated for the first time in the literature. Given their potential prognostic role and possible relevance as therapeutic targets, this study presents preliminary observations that add to the existing literature. Full article

Backgrounds: As one of the important liquid biopsy methods in recent years, circulating tumor cells (CTCs) have been proven to be valuable in judging metastasis and prognosis in various tumors. With the advent of immunotherapy, the expression of programmed death-ligand 1 (PD-L1) on CTCs has also attracted researchers’ attention, but its value in bladder cancer has not been fully explored. Methods: This study enrolled patients diagnosed with urothelial carcinoma who were treated in the Department of Urology, The First Affiliated Hospital of Nanjing Medical University from 2020 to 2023. Peripheral blood samples of the patients were collected to detect PD-L1+ CTCs. Meanwhile, the patients’ basic clinical characteristics, pathological data, and follow-up information were collected. The Kaplan–Meier method was used to estimate the overall survival (OS) and progression-free survival (PFS) of the patients, and the log-rank test was employed to evaluate the statistical differences. COX regression analysis and Firth penalized Cox regression analysis were adopted to assess the risk factors. Finally, according to the clinical or pathological characteristics, the effects of PD-L1+/PD-L1− CTCs on different subgroups of the population were evaluated, respectively. Results: According to the inclusion and exclusion criteria, a total of 109 patients were finally enrolled in this study, including 95 males and 16 females, with a median age of 68 years. The expression of PD-L1 on CTC was as follows: 19 patients were PD-L1+ CTC and 90 patients were PD-L1− CTCs. The results showed that the pathological grade of patients with PD-L1+ CTCs was significantly higher than that of patients with PD-L1− CTCs (p = 0.003). With a median follow-up time of 48 months (IQR: 45.8–49.5), prognostic analysis indicated that PD-L1+ CTCs were a risk factor for OS in bladder cancer patients (HR = 4.696 (1.477–13.596), p = 0.011). Subgroup analysis revealed that in patients with non-muscle-invasive bladder cancer (NMIBC) and in the subgroup of patients with high pathological grade, those with PD-L1+ CTCs exhibited significantly poorer prognosis in terms of PFS and OS compared with patients with PD-L1− CTCs. Conclusions: The presence of PD-L1+ CTCs in patients with bladder carcinoma may be closely associated with high-grade disease and poor OS. Full article

Background: Muscle-invasive bladder cancer (MIBC) is an aggressive disease with heterogeneous responses to neoadjuvant chemotherapy and emerging chemo-immunotherapy combinations. Reliable biomarkers to predict treatment responsiveness before therapy initiation are needed to guide patient selection. Objective: The objective of this study was to identify genomic and immune-related features associated with immune-active tumor phenotypes in MIBC using The Cancer Genome Atlas bladder cancer cohort (TCGA-BLCA). Methods: A retrospective bioinformatics analysis of TCGA-BLCA data was performed, evaluating gene expression, somatic mutations, tumor mutational burden (TMB), DNA damage response (DDR) gene status, and immune infiltration signatures. Immune enrichment metrics were derived from transcriptomic data. In the absence of direct treatment response data, a surrogate immune response classification was applied. Associations were analyzed using descriptive statistics and Firth’s penalized logistic regression. Results: Tumors classified as immune-high phenotype group based on immune-related features exhibited significantly higher global immune infiltration, including increased ImmuneScore and enrichment of cytotoxic and innate immune cells. In multivariable analysis, ImmuneScore was the only independent predictor of inferred responsiveness (p = 0.003). Conclusions: Global immune infiltration showed the strongest association with immune-active tumor phenotypes among the features examined in this TCGA-based analysis. These exploratory findings suggest that immune profiling may warrant further investigation as a component of tumor characterization in MIBC, pending validation in cohorts with clinical treatment and outcome data. Full article

Drug resistance remains a significant barrier to achieving durable treatment responses. Traditionally, resistance has been attributed to genetic alterations and clonal selection. However, accumulating evidence suggests that early adaptation to therapy is often mediated by non-genetic state transitions. In this review, we propose a conceptual framework in which resistance emerges through therapy-driven molecular evolution in bladder cancer, characterized by three interconnected axes: non-genetic plasticity, metabolic reorganization, and tumor microenvironment remodeling. Using the Gemcitabine-Resistant Cell (GRC) model as a temporal reference system, we describe a stepwise transition from drug-sensitive states dominated by proliferation to survival-optimized resistant states through a growth–survival trade-off. Early adaptive phases are marked by the attenuation of cell-cycle and glycolytic programs, increased epigenetic flexibility, and metabolic rewiring involving mitochondrial and lipid-associated pathways. Later phases involve the reinforcement of resistance through extracellular matrix remodeling, developmental and stress-response signaling, and immunometabolic interactions within the tumor microenvironment, including adenosine- and lipid-associated mediators. Projecting the GRC score onto a clinical bladder cancer cohort further suggests that these evolutionary patterns may also be reflected in patient tumors. Overall, this framework supports a temporally structured view of chemoresistance and highlights opportunities to therapeutically target transitional adaptive states before resistance becomes stabilized. Full article

Therapeutic cancer vaccines are a promising immunotherapy approach in genitourinary (GU) cancers, designed to stimulate antitumor immune responses through antigen-specific T-cell activation. Although agents such as bacillus Calmette–Guérin in bladder cancer and sipuleucel-T in prostate cancer have shown success, vaccine monotherapy has generally produced limited clinical benefit due to tumor heterogeneity, poor immune infiltration, and immunosuppressive tumor microenvironments. Multiple vaccine platforms have demonstrated safety and immunogenicity in prostate, renal cell, and urothelial cancers, but efficacy remains modest. Current strategies focus on multi-antigen targeting, improved antigen presentation, and combination therapies with immune checkpoint inhibitors, radiotherapy, and targeted agents to enhance antitumor activity. Advances in personalized vaccine design and delivery systems are driving progress, though challenges such as manufacturing complexity, cost, and biomarker development remain. Ongoing translational and clinical research will be critical to improving the effectiveness of vaccine-based immunotherapy in GU malignancies. Full article

Immune checkpoint receptors (ICRs) play a pivotal role in modulating antitumor immunity and have become central targets in the immunotherapy of genitourinary (GU) malignancies. This review provides a comprehensive overview of the fundamental mechanisms of ICR signaling, the expression and pathophysiological roles of these receptors in GU cancers (kidney, bladder, prostate, testicular, and penile), and the evolving therapeutic landscape. Key ICRs, including PD-1, CTLA-4, LAG-3, TIM-3, and TIGIT, orchestrate complex signaling cascades that can lead to T-cell exhaustion and tumor immune evasion. Their expression varies significantly across GU cancer types, histological subtypes, and tumor stages, influencing prognosis and therapeutic response. Immune checkpoint inhibitors (ICIs) reinvigorate antitumor immunity by disrupting these inhibitory pathways and remodeling the tumor microenvironment (TME); however, resistance mechanisms (primary, adaptive, and acquired) and immune-related adverse events (irAEs) pose significant clinical challenges. Established biomarkers such as PD-L1 expression, tumor mutational burden (TMB), and microsatellite instability (MSI)/deficient mismatch repair (dMMR) status guide ICI use, but their predictive power has limitations. Consequently, emerging tissue-based (e.g., immune cell signatures, multiplex IHC/IF, spatial transcriptomics), liquid biopsy-based (e.g., ctDNA, CTCs, exosomes), and imaging-based (radiomics, AI-driven analysis) biomarkers are under active investigation to refine patient selection and monitor treatment efficacy. The therapeutic armamentarium is rapidly expanding with novel ICIs targeting new receptors, bispecific antibodies, and innovative combination strategies involving ICIs with chemotherapy, targeted therapies, radiotherapy, and other immunotherapies. Furthermore, ICIs are increasingly explored in neoadjuvant, adjuvant, and maintenance settings. This review highlights the dynamic progress in understanding ICR biology and its clinical translation, emphasizing the ongoing efforts to develop more personalized and effective immunotherapeutic strategies for patients with genitourinary tumors. Full article

Background: Immunohistochemistry (IHC) plays a central role in subtyping of muscle-invasive bladder cancer (MIBC), yet conventional semi-quantitative scoring lacks objectivity and scalability. Automated digital pathology offers potential solutions, but requires robust, marker-specific validation against expert consensus scoring. Methods: We developed and internally validated an automated digital pathology pipeline for continuous IHC H-score quantification using QuPath (v0.6.0-arm64). Tissue microarrays (TMAs) were generated from transurethral resection of bladder tumor (TURBT) specimens from a cohort of patients with MIBC treated at Vancouver General Hospital. Cell detection was performed using StarDist (v0.9), followed by automated intensity-based H-score calculation for four epithelial IHC marker stains (CK14, CK20, CK5/6, and Uroplakin II). H-scoring was then restricted to tumor epithelium by object-level classification using a supervised tumor/non-tumor classifier trained on pathologist-reviewed annotations. Automated scores were compared with consensus scores from three blinded pathologists using Pearson correlation, linear regression, intraclass correlation coefficients (ICC), and Bland–Altman analysis. Results: Automated H-scores demonstrated strong agreement with pathologist consensus across all four markers. CK14 showed near-perfect agreement (ICC ≈ 0.99) with minimal bias and narrow limits of agreement. CK20 also showed high agreement (ICC ≈ 0.95). CK5/6 and Uroplakin II demonstrated slightly lower agreement (ICC ≈ 0.92 to 0.93) with mild proportional bias. Across markers, the automated pipeline preserved a broad H-score range, with range ratios of 0.96 to 0.99. Conclusions: This study establishes a robust, methods-forward pipeline for automated continuous IHC H-scoring in MIBC. The internally validated framework provides a scalable foundation for external cohort testing and future clinical outcome-associated biomarker analyses. Full article