High-grade Urothelial Bladder Cancer Research Articles

Adenoid cystic/basal-cell carcinoma of the prostate following high-grade urothelial bladder cancer: a case report

Adenoid cystic/basal-cell carcinoma of the prostate following high-grade urothelial bladder cancer: a case report

A Comprehensive Analysis of Immune Response in Patients with Non-Muscle-Invasive Bladder Cancer

Bladder carcinoma has elevated morbimortality due to its high recurrence and progression in localized disease. A better understanding of the role of the tumor microenvironment in carcinogenesis and response to treatment is needed. Peripheral blood and samples of urothelial bladder cancer and adjacent healthy urothelial tissue were collected from 41 patients and stratified in low- and high-grade urothelial bladder cancer, excluding muscular infiltration or carcinoma in situ. Mononuclear cells were isolated and labeled for flow cytometry analysis with antibodies aimed at identifying specific subpopulations within T lymphocytes, myeloid cells and NK cells. In peripheral blood and tumor samples, we detected different percentages of CD4+ and CD8+ lymphocytes, monocyte and myeloid-derived suppressor cells, as well as differential expression of activation- and exhaustion-related markers. Conversely, only a significant increase in bladder total monocytes was found when comparing bladder and tumor samples. Interestingly, we identified specific markers differentially expressed in the peripheral blood of patients with different outcomes. The analysis of host immune response in patients with NMIBC may help to identify specific markers that allow optimizing therapy and patient follow-up. Further investigation is needed to establish a strong predictive model.

Impact of bacillus Calmette-Guerin intravesical therapy on the diagnostic efficacy of The Paris System for Reporting Urinary Cytology in patients with high-grade bladder cancer.

In high-grade urothelial carcinoma (UC) of the bladder, bacillus Calmette-Guerin (BCG) therapy is a therapeutic mainstay, and urinary cytology is recommended to detect recurrences. However, intravesical BCG instillations can induce morphologic changes in urothelial cells. The authors investigated the impact of BCG therapy on the efficacy of urinary cytology. Matched pathology and cytology samples from patients undergoing transurethral resection of the bladder after BCG therapy were assessed. Cytology samples were graded according to The Paris System for Reporting Urinary Cytology. Diagnostic quality criteria were tested for different cutoff definitions, and the results were compared between those obtained <100 versus ≥100 days after the last BCG instillation. In addition, the oncologic outcome of false-positive results was assessed. In total, 389 matched cases from 197 patients who had a history of high-grade UC (HGUC) were identified. Sixty cases (15.7%) were diagnosed as high-grade urothelial bladder cancer. The cytology diagnoses were as follows: non-HGUC, 191 cases (49.1%); atypical urothelial cells, 80 cases (20.6%); suspicious for HGUC, 56 cases (14.4%); and HGUC, 56 cases (14.4%). Interrater reliability was substantial (κ = 0.660). Sensitivity increased from 45% to 75% when cases diagnosed as suspicious for HGUC were also counted as positive. Notably, sensitivity was reduced within the first 100 days after BCG therapy (61.9%) compared with sensitivity at longer intervals (82.1%). Reactive atypia (odds ratio, 4.155; 95% confidence interval, 2.136-8.085; P < .001) and cellular degeneration (odds ratio, 5.050; 95% CI, 2.094-12.175; P < .001) of urothelial cells were associated with false-positive rates, and 44.7% of patients who had a false-positive cytology classification presented with HGUC during follow-up. BCG therapy has a short-term adverse impact on the efficacy of urinary cytology. After BCG therapy, cases classified as suspicious for HGUC should be considered positive. Importantly, patients with false-positive cytology findings should be closely monitored.

PD03-03 HOW EFFECTIVE IS MULTIPARAMETRIC MAGNETIC RESONANCE IMAGING VESICAL IMAGING REPORTING AND DATA SYSTEM (VI-RADS) FOR BLADDER CANCER STAGING: CAN IT REPLACE RE STAGING TRANSURETHRAL RESECTION OF BLADDER CANCER IN BLADDER CANCER?

You have accessJournal of UrologyBladder Cancer: Non-invasive I (PD03)1 Apr 2020PD03-03 HOW EFFECTIVE IS MULTIPARAMETRIC MAGNETIC RESONANCE IMAGING VESICAL IMAGING REPORTING AND DATA SYSTEM (VI-RADS) FOR BLADDER CANCER STAGING: CAN IT REPLACE RE STAGING TRANSURETHRAL RESECTION OF BLADDER CANCER IN BLADDER CANCER? Muhammad Abdus Salam*, Sharmin Akther, Faisal Islam, Pranashish Saha, Ibrahim Kaiser, Ghazi Shahinul Islam, Shahed Hossain, and SM Bakhtiar Muhammad Abdus Salam*Muhammad Abdus Salam* More articles by this author , Sharmin AktherSharmin Akther More articles by this author , Faisal IslamFaisal Islam More articles by this author , Pranashish SahaPranashish Saha More articles by this author , Ibrahim KaiserIbrahim Kaiser More articles by this author , Ghazi Shahinul IslamGhazi Shahinul Islam More articles by this author , Shahed HossainShahed Hossain More articles by this author , and SM BakhtiarSM Bakhtiar More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000000823.03AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Multiparametric MRI (mpMRI) is widely used in the imaging of other types of cancers, though its usefulness in Urothelial cancer of bladder has yet to be fully elucidated. We wanted to determine the efficacy of staging accuracy of routine use of mpMRI and VI-RADS (Vesical Imaging-Reporting And Data System) in staging Urothelial bladder cancer. METHODS: This prospective study was undertaken on 62 patients with mpMRI and VI-RADS in staging high grade Urothelial bladder cancer. Primary Transurethral resection of bladder tumor ( TURBT) mpMRI was performed routinely in all cases, from Jan 2018 to July 2019. Of them 36 patients with High grade Urothelial carcinoma of bladder, the specimen did not contain Muscularis Propria requiring a Restaging TURBT. Radical cystectomy was performed in 18 patients during this time frame. The mpMRI staging was correlated with the final pathologic staging. RESULTS: Analysis of our results revealed a highly significant (P<-0.001) mpMRI accuracy of 88% for muscle invasion, 96% accuracy for perivesical fat invasion and 74% accuracy of MRI detection of lymph node metastases. CONCLUSIONS: A larger prospective study is however, required for determining the exact role of MRI in the staging of bladder carcinoma. Source of Funding: No Sponsor © 2020 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 203Issue Supplement 4April 2020Page: e72-e73 Advertisement Copyright & Permissions© 2020 by American Urological Association Education and Research, Inc.MetricsAuthor Information Muhammad Abdus Salam* More articles by this author Sharmin Akther More articles by this author Faisal Islam More articles by this author Pranashish Saha More articles by this author Ibrahim Kaiser More articles by this author Ghazi Shahinul Islam More articles by this author Shahed Hossain More articles by this author SM Bakhtiar More articles by this author Expand All Advertisement PDF downloadLoading ...

Second Transurethral Resection of Bladder Tumor: Is it Necessary in All T1 and/or High-Grade Tumors?

To evaluate the role of second transurethral resection of bladder tumor (TURBT) in patients with T1 and/or high-grade bladder tumor regarding tumor size, multiplicity, and presence or absence of muscle in specimens of initial resection. A total of 107 patients with either primary T1 or high-grade urothelial bladder cancer underwent second TURBT within 6 weeks after initial surgery and prior to starting intravesical immunotherapy. We assessed the incidence of residual disease and upstaging in second TURBT. Upstaging was noted in 11 (10.3%) patients and residual tumor was evident in 29 (27%) patients. Disease upstaging had a statistically significant association with tumor size, multifocality, and absence of muscle at initial resection in univariate analysis. Presence of residual tumor in second resection also showed significant association with tumor size and absence of muscle at initial resection but not multifocality. Multivariate logistic regression analysis revealed that absence of muscle at initial resection independently predicts disease upstaging during second TURBT (OR = 8.123, 95% CI: 1.478-44.632). Furthermore, both tumor size (OR = 13.573, 95% CI: 3.104-59.359) and absence of muscle (OR = 21.214, 95% CI: 6.062-74.244) were independent predictors of residual disease in second TURBT. We showed that second TURBT in a subset of patients with single, small T1 and/or high-grade tumor who underwent complete initial resection might be of limited value.

CCDC6 and USP7 expression levels suggest novel treatment options in high-grade urothelial bladder cancer

BackgroundThe muscle invasive form of urothelial bladder cancer (UBC) is a deadly disease. Currently, the therapeutic approach of UBC is mostly based on surgery and standard chemotherapy.Biomarkers to establish appropriate drugs usage are missing. Deficiency of the tumor suppressor CCDC6 determines PARP-inhibitor sensitivity. The CCDC6 levels are modulated by the deubiquitinase USP7. In this work we scored CCDC6 and USP7 expression levels in primary UBC and we evaluated the expression levels of CCDC6 in correlation with the effects of the PARP-inhibitors combined with the USP7 inhibitor, P5091, in vitro. Since PARP-inhibitors could be enhanced by conventional chemotherapy or DNA damage inducers, we tested the new agent RRx-001, able to induce DNA damage, to prove the benefit of combined treatments in bladder cancer cells.MethodsThe J82, T24, 5637 and KU-19-19 bladder cancer cells were exposed to USP7 inhibitor P5091 in presence of cycloheximide to analyse the CCDC6 stability. Upon the CCDC6 degradation induced by P5091, the cells sensitivity to PARP-inhibitor was evaluated by cell viability assays. The ability of the DNA damage inducer RRx-001 to modulate CCDC6 protein levels and H2AX phosphorylation was detected at immunoblot. The combination of USP7 inhibitor plus RRx-001 enhanced the PARP-inhibitor sensitivity, as evaluated by cell viability assays. The results of the scores and correlation of CCDC6 and USP7 expression levels obtained by UBC primary biopsies staining were used to cluster patients by a K-mean cluster analysis.ResultsP5091 determining CCDC6 degradation promoted bladder cancer cells sensitivity to PARP-inhibitor drugs. RRx-001, by inducing DNA damage, enhanced the effects of the combined treatment. The immunohistochemical staining of both CCDC6 and USP7 proteins allowed to cluster the high grade (G3) UBC patients, on the basis of CCDC6 expression levels.ConclusionsIn high grade UBC the identification of two clusters of patients based on CCDC6 and USP7 expession can possibly indicate the use of PARP-inhibitor drugs, in combination with USP7 inhibitor in addition to the DNA damage inducer RRx-001, that also acts as an immunomodulatory agent, offering novel therapeutic strategy for personalized medicine in bladder cancer patients.

Safety and Efficacy of Hypofractionated Radiotherapy With Capecitabine in Elderly Patients With Urothelial Carcinoma.

Bladder cancer is commonly diagnosed in patients ineligible for radical cystectomy or chemoradiotherapy (chemo-RT) with cisplatin or fluorouracil with mitomycin. We assessed tolerability, efficacy, and toxicity of hypofractionated radiotherapy with capecitabine in this challenging population. Patients with high-grade urothelial bladder cancer ineligible for radical cystectomy or high-intensity chemo-RT underwent maximal transurethral resection of bladder tumor followed by capecitabine (median, 825 mg/m2 per day 2 times a day) and radiation (median, 55 Gy in 2.2 Gy per fraction). Patients underwent surveillance cystoscopy and imaging, and were evaluated for toxicity, freedom from local failure and freedom from distant metastasis, progression-free survival, and overall survival. Eleven patients (median age, 80 years) with localized disease (n= 7), locally advanced disease (n= 3), or local-only recurrence after cystectomy (n= 1) were treated. Four patients (35%) had an Eastern Cooperative Oncology Group performance status of 2; median Charlson comorbidity index was 5. There was 1 acute grade 3 genitourinary event (9%), 6 acute grade 3 hematologic events (55%) of lymphopenia, and no acute grade 4 or higher events or hospitalizations. Ten patients (91%) completed radiotherapy, while 4 patients (36%) temporarily discontinued capecitabine. The complete response rate in the bladder was 64%. Two patients (18%) experienced late grade 1/2 genitourinary toxicities, and 1 (9%) experienced a transient late grade 4 genitourinary toxicity. With a median follow-up of 16.6 months, overall survival, progression-free survival, freedom from local failure, and freedom from distant metastasis at 1 year were 82%, 55%, 100%, and 55%, respectively, and at 2 years were 61%, 41%, 80%, and 55%, respectively. Hypofractionated chemo-RT was well tolerated and was associated with a high rate of local control in this comorbid population, thus providing a treatment option for select bladder cancer patients.

Diabetic Ketoacidosis Triggered by Pembrolizumab in a Patient with Bladder Cancer

This is a 71-year-old male with history of Hypertension, well controlled type 2 DM on metformin (HbA1c 6.6%), and high grade urothelial bladder cancer treated with transurethral resection of the bladder. Due to cancer progression, he required neoadjuvant chemotherapy followed by Pembrolizumab in July 2017. He presented with fatigue, polyuria and polydipsia in August 2017. He was diagnosed with DKA and managed in the ICU. Workup revealed HbA1c 9%, high GAD-65 antibody (&amp;gt;250 IU/ML), and low C-peptide (0.26 ng/ml). He was stabilized and discharged on insulin. T cell activity is regulated by programmed death ligand-1(PD-L1)/programmed death-1 (PD-1). Tumor cells select specific immune-checkpoint pathways as a mechanism of immune resistance against T cells and evade the antitumor immune response by overexpressing PD-L1. Pembrolizumab is a humanized monoclonal antibody against PD-1 acting as an immune checkpoint inhibitor (ICI). Pembrolizumab is approved for treating metastatic urothelial carcinoma, cisplatin-ineligible cancer or disease progression after cisplatin therapy. ICIs are associated with multi-system immune related adverse events (irAEs) due to the disinhibition of the host immune homeostasis. Most of the irAEs are reversible with supportive treatment, however life-threatening irAEs can occur including severe skin reaction, type 1 DM, hypophysitis, and rhabdomyolysis. In our patient, we suspected pembrolizumab was related to new onset type 1 DM. type 1 DM and DKA were reported in 6 (0.2%) of 2,799 patients on Pembrolizumab in 3 randomized, open-label, and active-controlled clinical trials. There are 12 reported cases of PD-1/PD-L1-associated type 1 DM. The onset of disease from use varied from 1 week to 12 months, with most patients presenting with type 1 DM with DKA. ICIs are breakthroughs in cancer treatment. Clinicians should be aware of their possible irAEs. The development of autoimmune diabetes is rare, but its complications are life threatening if not attended to immediately. Disclosure T. Al Shamy: None. M. Aguasvivas: None. M. Serhan: None. M.M. Fojas: None.

Infiltrative lamina propria invasion pattern as an independent predictor for cancer-specific and overall survival of instillation treatment-naïve stageT1 high-grade urothelial bladder cancer.

To investigate established prognostic factors and relatively new histopathological tumor characteristics including metric substage and lamina propria invasion patterns in a large series of T1 high-grade non-muscle-invasive bladder cancer. Between 1989 and 2012, 322 patients with initial stageT1 high-grade bladder cancer underwent transurethral resection, followed by re-transurethral resection and a conservative approach with follow-up regime alone or instillation treatment. Transurethral resection specimens were reassessed by two experienced urological pathologists for tumor grade according to the World Health Organization 1973 classification, metric T1 substage, lamina propria invasion pattern and associated carcinoma insitu. The median follow-up period was 42months (interquartile range 25-72months). In addition to Kaplan-Meier analyses, uni- and multivariable Cox regression analyses were used to compare progression-free survival, cancer-specific survival and overall survival for the studied parameters comparing two subcohorts. While in patients after instillation treatment no examined feature was shown as an independent predictor for prognosis, there were predictive histopathological features in multivariable Cox regression analyses in instillation treatment-naïve patients: associated carcinoma insitu (hazard ratio 2.278, 95% confidence interval 1.119-4.634, P=0.023) and World Health Organization 1973 grade3 (hazard ratio 2.950, 95% confidence interval 1.021-8.536, P=0.046) for worse progression-free survival, infiltrative lamina propria tumor pattern for worse cancer-specific survival (hazard ratio 2.369, 95% confidence interval 1.034-5.429, P=0.042) and overall survival (hazard ratio 1.049, 95% confidence interval 1.024-1.075, P=0.001). The results of the present T1 high-grade bladder cancer series suggest that lamina propria invasion pattern is a promising parameter to predict the prognosis of T1 high-grade bladder cancer in an instillation treatment-naïve subcohort. Prospective multicenter evaluations are warranted. The need for instillation treatment in T1 high-grade bladder cancer is clearly demanded.

Is there a role for second transurethral resection in pTa high-grade urothelial bladder cancer?

IntroductionEvidence for second transurethral resection of bladder tumour (TURBT) for pTa high-grade lesions is limited. This study aims to examine the role of a second TURBT in the pTa high-grade group and to generate recurrence and progression data for this group.Material and methodsWe retrospectively studied the clinical profiles and outcomes of all patients diagnosed with high-grade pTa lesions at first TURBT, between the years 2006–2015. Firstly, in patients who underwent a complete first TURBT, we calculated the proportion of patients with positive findings on second TURBT. Secondly, we assessed whether those who underwent a second TURBT had a longer recurrence-free survival compared to those who underwent a single TURBT.ResultsOne hundred and twelve patients had a pTa high-grade urothelial bladder tumor (WHO 2004 classification) at first TURBT, out of whom 43 (38.3%) had a second TURBT. Indications for second TURBT were high-grade lesions (n = 36), absence of detrusor muscle (n = 2), and incomplete resection (n = 5). Out of the 36 patients who had a complete first TURBT and underwent a second look TURBT, 7 patients had positive findings (3 carcinoma in situ, 2 pTa low-grade lesions and 2 pTa high-grade lesions) and there was no upstaging. Of the 5 patients with an incomplete first TURBT, one upstaged to pT1 on second TURBT. Of the 81 patients who followed up with us, 25.9% had a recurrence and 8.6% progressed. The estimated median recurrence free survival was 60 months (95% CI 29.2–90.7) for the whole group and 76 months vs. 45 months for the second and single TURBT group respectively – a difference that was clinically, though not statistically, significant. Multiple (≥2) tumours had a lower recurrence free survival (HR of 4.60, CI 1.67-12.63, p = 0.003).ConclusionsOf the patients with pTa high-grade tumours who had a second TURBT after a complete first TURBT, 19.4% had a positive finding. Multiple tumours are four times as likely to recur as solitary tumours. The role of a second TURBT in this group needs to be studied in larger patient cohorts before a recommendation regarding its lack of clinical utility can be made conclusively.

Opium and grade of urothelial bladder cancer

Background: Bladder cancer is a serious global problem. Although the risk factors of bladder cancer have been found through many studies, the effect of them on the grade of this cancer has not been focused previously. Objectives: This study was conducted to evaluate the association of some risk factors of urothelial bladder cancer on its grade. Patients and Methods: The patients with proved diagnosis of urothelial carcinoma of bladder were enrolled in this study. They were divided into case group (those with high-grade bladder cancer) and control group (those with low-grade bladder cancer). Each of case and control groups contained 103 patients. Age, gender, place of residence, cigarette smoking state, opium consumption state, occupation, previous exposure to poisons or chemicals, and family history of bladder cancer were compared between the case and control groups. Results: Eighty-two patients (79.6%) of the case group and 87(84.5%) of the control group were male. The mean age of the case group was significantly higher than control group (66.1±11.8 and 61.7±14.2, respectively; P=0.003). The results of this study showed that ever usage of opium was significantly higher in the case group (OR=2.97; 95% CI: 1.64- 5.38; P <0.001). Although mean pack-years of cigarette smoking of case group was significantly higher than control group in univariate analysis (P=0.009), it was omitted from regression models when multivariate analysis was applied. Conclusions: High grade urothelial bladder cancer has been significantly associated with opium usage and older ages

Abstract 5120: The novel claudin-low molecular subtype of high-grade urothelial bladder cancer is highly immunogenic yet immunosuppressed

Abstract Rationale: High-grade urothelial carcinoma of the bladder is a heterogeneous disease, with molecular subtypes characterized by distinct tumor biologies and prognoses. Our group and others have described the basal and luminal subtypes, and we now report on the discovery of the claudin-low subtype, all of which resemble analogous subtypes in breast cancer. Methods: We analyzed mRNA sequencing and whole exome sequencing data for The Cancer Genome Atlas (TCGA) bladder tumors and tumors collected at UNC. We performed unsupervised hierarchical clustering on relative gene expression values with significance testing using SigClust to identify the claudin-low subtype. Basal, luminal, and claudin-low bladder tumors were compared for enrichment of genetic features (single nucleotide and copy number variation), gene set expression, immune gene signature expression, T cell receptor (TCR) and B cell receptor (BCR) gene segment expression, and number of predicted MHC Class I and Class II neoantigens. We further report on the first use of our VDJician software to reconstruct full-length rearranged BCR sequences from short-read RNA sequencing data in bladder cancer. Results: Claudin-low bladder tumors were defined by low expression of tight junction claudin-proteins, high expression of epithelial-to-mesenchymal transition genes and immune gene signatures, and were associated with reduced overall survival compared to luminal tumors. A minimal gene set classifier to identify claudin-low tumors showed some but not complete overlap with an optimal classifier derived in breast cancer. Claudin-low bladder tumors were enriched for multiple genetic features: increased rates of RB1, EP300, and NCOR1 mutations, increased frequency of EGFR amplification, decreased rates of FGFR3, ELF3, and KDM6A mutations, and decreased frequency of PPARG amplification. Claudin-low tumors showed the highest expression of immune gene signatures (including an immunosuppression signature), however in contrast to basal tumors, increased immune gene signature expression was not associated with prolonged survival. Claudin-low tumors also showed the highest overall expression of rearranged BCR sequences but lowest BCR repertoire diversity. Predicted neoantigen burden did not vary significantly by subtype, however broad cytokine and chemokine expression levels were elevated in claudin-low tumors, potentially related to low PPARG activity driving increased NFKB activity. Conclusions: Claudin-low bladder cancer is a novel molecular subtype with distinct molecular and immunologic features and prognostic significance. Given the presence of dense immune infiltrates, BCR repertoire characteristics consistent with an antigen-driven response, and high expression of immunosuppression genes, claudin-low bladder tumors may be primed to respond to immune checkpoint inhibitor therapy. Citation Format: Benjamin G. Vincent, William Kim, Jordan Kardos, Shengjie Chai, Joel Parker, Lisle Mose, Sara Selitsky, Michael Iglesia, Matthew Milowsky. The novel claudin-low molecular subtype of high-grade urothelial bladder cancer is highly immunogenic yet immunosuppressed. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5120.

High-Grade Urothelial Bladder Cancer in Children: A Case Report and Critical Analysis of the Literature

Bladder cancer is relatively common in adults; however, in children, bladder malignancies are extremely rare. In the present case report, we describe the diagnostic, therapeutic, and follow-up management of bladder cancer in a 3-year-old boy. Furthermore, we focus on typical and less frequently seen histological characteristics of bladder cancer in childhood. Despite its low incidence and prevalence, it is very important to emphasize that bladder cancer is the most serious condition which should not be missed in the differential diagnosis of hematuria or urinary tract infection.

Genomic Predictors of Survival in Patients with High-grade Urothelial Carcinoma of the Bladder

Urothelial carcinoma of the bladder (UCB) is genomically heterogeneous, with frequent alterations in genes regulating chromatin state, cell cycle control, and receptor kinase signaling. To identify prognostic genomic markers in high-grade UCB, we used capture-based massively parallel sequencing to analyze 109 tumors. Mutations were detected in 240 genes, with 23 genes mutated in ≥5% of cases. The presence of a recurrent phosphatidylinositol-4,5-bisphosphate 3-kinase, catalytic subunit alpha (PIK3CA) mutation was associated with improved recurrence-free survival (RFS) (hazard ratio [HR]: 0.35; p=0.014) and improved cancer-specific survival (CSS) (HR: 0.35; p=0.040) in patients treated with radical cystectomy (RC). In multivariable analyses controlling for pT and pN stages, PIK3CA mutation remained associated with RFS (HR: 0.39; p=0.032). The most frequent alteration, TP53 mutation (57%), was more common in extravesical disease (69% vs 32%, p=0.005) and lymph node–positive disease (77% vs 56%, p=0.025). Patients with cyclin-dependent kinase inhibitor 2A (CDKN2A)–altered tumors experienced worse RFS (HR: 5.76; p<0.001) and worse CSS (HR: 2.94; p=0.029) in multivariable analyses. Mutations in chromatin-modifying genes were highly prevalent but not associated with outcomes. In UCB patients treated with RC, PIK3CA mutations are associated with favorable outcomes, whereas TP53 and CDKN2A alterations are associated with poor outcomes. Genomic profiling may aid in the identification of UCB patients at highest risk following RC. Patient summaryUsing next-generation sequencing, we identified genomic subsets of high-grade urothelial bladder cancer associated with favorable and unfavorable outcomes. These findings may aid in the selection of patients most likely to benefit from novel combined modality approaches.

Promoter hypermethylation of tumor suppressor genes correlates with tumor grade and invasiveness in patients with urothelial bladder cancer.

PurposeTo investigate the promoter methylation status at selected loci which encode for key proteins involved in apoptosis, DNA repair, cell cycle control and progression in urothelial cell carcinoma of bladder and compare the findings from tissue samples with that of plasma.MethodsTotal genomic DNA was isolated from 43 non-muscle invasive (low grade) and 33 muscle invasive (high grade) urothelial bladder cancer samples along with 10 control cases of normal bladder mucosa. Promoter methylation status was investigated for RASSF1A, APC, MGMT, CDKN2A and CDKN2B genes using real-time methylation-specific PCR with SYBR® green. Plasma samples from 16 patients with muscle invasive high grade bladder cancer were also subjected to similar analyses.ResultsPromoter hypermethylation was frequently observed in RASSF1A, APC and MGMT gene promoters (p-value < 0.001). The methylation was more prominent in the muscle invasive high grade bladder cancer when compared to non-muscle invasive low grade group (p-value < 0.001) and normal bladder mucosa (p-value < 0.05). The RNA expression of RASSF1A, APC and MGMT was also found to be decreased in the muscle-invasive high grade bladder cancer when compared to the non muscle invasive low grade group (p-value < 0.05). RASSF1A, MGMT and CDKN2A showed comparable results when data from 16 plasma samples was compared to the corresponding tissue samples.ConclusionOur results suggest that epigenetic silencing of RASSF1A, APC and MGMT genes is strongly associated with invasive high grade urothelial bladder cancer. Thus, status of promoter methylation has the potential to serve as valuable tool for assessing aggressiveness of urothelial cell carcinoma of bladder.Electronic supplementary materialThe online version of this article (doi:10.1186/2193-1801-3-178) contains supplementary material, which is available to authorized users.

Expression of histone deacetylases 1, 2 and 3 in urothelial bladder cancer

BackgroundHistone deacetylases (HDACs) are known to be associated with an overexpression in different types of cancer such as colon and prostate cancer. In this study we aimed to evaluate the protein expression of class I HDACs in urothelial carcinoma of the bladder.MethodsA tissue microarray containing 348 tissuesamples from 174 patients with a primary urothelial carcinoma of the bladder was immunohistochemically stained for HDAC 1, 2 and 3. Intensity of staining was evaluated and the association with clinico-pathological features and prognosis was assessed.ResultsHigh HDAC expression levels were found in 40 to 60% of all investigated urothelial carcinomas (HDAC-1: 40%, HDAC-2: 42%, HDAC-3: 59%).HDAC-1 and HDAC-2 were significantly associated with higher tumour grades.Although all three markers could not predict progression in univariate analyses, high HDAC-1 expression was associated with a trend toward poorer prognosis. Patients with high-grade tumours and high expression levels of HDAC-1 were more likely to progress compared to all other patients (p < 0.05).ConclusionsHigh-grade noninvasive papillary bladder tumours are associated with high expression levels of HDAC-1 and HDAC-2. High grade tumours in combination with high expression of HDAC-1 showed a worse prognosis than the other tumours. The high expression levels of HDACs observed particularly in high grade urothelial bladder cancer clearly warrant subsequent studies on the potential use of HDAC inhibitors as a novel therapeutic approach.

Abstract 442: Increased miR-let7c level indicates transition from localized bladder cancer to metastasis

Abstract Purpose/Objective(s): MicorRNAs are a group of short ribonucleic acid molecules with an average of 22 nucleotides and play an important role in regulation of gene expression. Because the microRNA profiles of tumor are related to cancer development and prognostication, it becomes a new frontier of cancer research. The current tools for studying microRNA like microarray have limited dynamic range and poor sensitivity, and Taqman microRNA assay has higher sensitivity but limited number of microRNAs. The SOLiD system provides a sensitive and robust assay for discovery and profiling of novel microRNAs using sequencing by ligation. The aim of this study is to find key miRNA which can provide insights into the cancer cell invasion and metastasis. Materials/Methods: 12 bladder cancer FFPE samples were used in this study, including 6 cases of stage N0, 3 cases of stage N1 and 3 cases of stage N2. Total RNA from FFPE blocks was isolated by Ambion RecoverAll Total Nucleic Acid Isolation kit according to manufacturer's instructions. The concentration and quality of total RNA were examined by Nanodrop 1000 spectrophotometer. The small RNA was enriched by flashPAGE fractionation and the microRNA library was built using Ambion SOLiD Small RNA Expression Kit with the barcode system (Life Technologies, CA, US). 35 bp long reads were generated on the SOLiD (version 3.0) sequencing system (Life Technologies) and were analyzed by using the SOLiD System Small RNA Analysis Tool (Life Technologies). Results: Analyzed samples were from male caucasian patients with a median age of 63.5 years. Tumors were all grade 3 urothelial carcinomas. Patients had T1-4 and M0-1 disease. We examined global miRNA expression in bladder cancer FFPE samples, and SOLiD results revealed that the expression of 10 microRNAs significantly differed between N0 stage and N1/2 stage. miR-let7c precursor and mature miR-let7c were upregulated in samples from patients with lymphnode metastasis. A previous study from Leite et al. showed that miR-let7c expression was similar in matched FFPE and frozen prostate cancer tissue with a Pearson's correlation coefficient of 0.927. Conclusion: miR-let7c expression is associated with the presence of lymphnode metastasis in high-grade urothelial bladder cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 442. doi:1538-7445.AM2012-442

Evaluation of Urine Tumor-Associated Trypsin Inhibitor, CYFRA 21-1, and Urinary Bladder Cancer Antigen for Detection of High-Grade Bladder Carcinoma

To assess the value of urine tumor-associated trypsin inhibitor (TATI), CYFRA 21-1, which measures cytokeratin 19 fragment, and urinary bladder carcinoma antigen (UBC) for the detection of high-grade bladder carcinoma. A total of 160 individuals were enrolled in the present study. Of these, 80 were patients with proven primary high-grade urothelial bladder cancer (group 1), 40 were healthy volunteers (group 2), and 40 had history of benign urologic disease (group 3). All were evaluated with respect to urinary TATI, CYFRA 21-1, and UBC levels. All these markers were evaluated using commercial kits. Cytology was also performed. The TATI measurements were significant greater in group 1 compared with groups 2 and 3. The cutoff point used for TATI, CYFRA 21-1, and UBC was 22, 2.8, and 12 microg/L, respectively. The overall sensitivity was 85.7% for TATI, 61.9% for CYFRA 21-1, 50% for UBC, and 42.8% for cytology. TATI was significantly more sensitive in Stage Ta (80%) than was CYFRA 21-1 (32%), UBC (12%), and cytology (20%). TATI was also more sensitive compared with other tumor markers for Stage T1 but not for Stage T2 or T3. The results of our study have shown that TATI is a promising urinary tumor marker for high-grade urothelial bladder cancer. It is more sensitive than CYFRA 21-1, UBC, and cytology for Stage Ta and T1 bladder cancer.

Immunohistochemical profile of high-grade urothelial bladder carcinoma and prostate adenocarcinoma

The differential diagnosis between poorly differentiated prostate adenocarcinoma (PAC) involving the bladder and high-grade urothelial bladder cancer (UC) with prostate extension can be very challenging. The aim of this study is to evaluate the use of a panel of antibodies to distinguish the poorly differentiated forms of these two tumors. We evaluated a series of 40 PAC cases (Gleason's grade ≥ 8) and 45 (G3) UC cases obtained from transurethral endoscopic resection material. Immunohistochemical analysis was performed using the following antibodies: prostate acid phosphatase (PAP), prostate-specific antigen (PSA), uroplakin III (UP), thrombomodulin (TM), cytokeratin (CK) 7, and CK20. PAC expressed PSA and PAP in 34 and 38 cases, respectively. The sensitivity and specificity of expressing at least 1 marker (PSA+ or PAP+) is 95% and 100%, respectively. All UC cases were negative for both markers. UC expressed UP and TM in 27 and 22 cases, respectively. In addition, 36 of 45 cases stained positively for at least 1 marker (UP + or TM +) with specificity and sensitivity of 80% and 100%, respectively. All cases of PAC were negative for both markers. Twenty-eight UC cases were CK7+/CK20 +, and 4 PAC cases stained positively for both markers. On the other hand, 29 PAC cases and 4 UC cases were CK7-/CK20-. We concluded that PSA, PAP, UP, and TM are very useful markers in differentiating poorly differentiated UC from PAC. Finally, when all 4 markers (PAP, PSA, UP, and TM) were negative, CK7 and CK20 appeared of no major use in making the differential diagnosis. HUM PATHOL 33:1136-1140. Copyright 2002, Elsevier Science (USA). All rights reserved.