Grade Non-muscle Invasive Bladder Cancer Research Articles

Digital holography can differentiate between bladder cancer grades.

413 Background: Treatment of high grade non-muscle invasive bladder cancer (HG NMIBC) is controversial with BCG instillation and early cystectomy as viable options. However, accurate predictive markers for HG NMIBC are lacking. Refractive index (RI) is an optical feature of tissues and has been shown to be correlated with various oncologic outcomes in prostate and breast cancers. The role of RI in NMIBC has not been explored yet. The aim of this study is to measure and compare RI values of benign and malignant low and high grade NMIBC. Methods: After IRB approval, tissue samples were obtained from high and low grade NMIBC patients. Measurements were performed on non-stained slides using digital holography. Areas of benign and malignant tissues were marked by a fellowship trained GU pathologist. The average RI (RIavg) and RI variance (RIvar) over malignant and benign cell nuclei were measured. Comparison between benign and malignant tissue and between low and high grade tumors was performed with the Student's t-test. Results: Multiple nuclei of LG and HG patients were measured . Mean difference of RIavg between malignant and benign cells was 0.03 in HG patients (p < 0.001). Mean difference of RIvar between malignant and benign tissues was 0.016 in HG patients (p < 0.001). In LG tissues, there was no significant difference in RIavg or RIvar between benign and malignant cells. Finally, there was a significant difference in RIavg between HG and LG cells (p < 0.001). Conclusions: For the first time, RI was measured in bladder cancer tissue by digital holography. The RI values and variance were different between malignant and benign cells in HG tissues. Furtheremore, we found a significant difference in RI values between HG and LG cells. This technology may assist in the future in tissue diagnosis and classification of bladder tumor.

The rational and benefits of the second look transurethral resection of the bladder for T1 high grade bladder cancer.

The second look transurethral resection of the bladder (TURB) represents a fundamental step in the treatment of papillary non-muscle invasive bladder cancer (NMIBC); it is therefore recommended by all guidelines. However, not all the literature agrees on its staging value and its ability to improve oncological outcomes of patients. Therefore, the purpose of this review is to evaluate the strengths and weaknesses of second look TURB, trying to depict its evolving role in the management of high grade NMIBC. Using Medline, a non-systematic review was performed including articles between January 2000 and June 2018. English language original articles, reviews and editorials were selected based on their clinical relevance. To date, TURB seems to be largely inadequate in retrieving a correct diagnosis and in removing all tumor tissue. Second look TURB maximizes staging accuracy, allows to clear residual cancer and yields prognostic advantages allowing key information to identify possible candidates for immediate radical cystectomy for very high risk T1HG tumors. Moreover, it seems to have a therapeutic benefit by improving recurrence- and progression-free survivals. However, few recent large studies showed that these advantages seem to be limited to patients without detrusor muscle present at first resection. Similarly, the presence of residual disease and the risk of upstaging are related to the presence of detrusor muscle in specimen. It could well be that in the future the presence of detrusor muscle would be a quality criteria to avoid an unnecessary second look TURB as shown by recent studies using the en-bloc resection technique. Finally, it has to be underlined that this is a surgical procedure not free of risks and complications and with a non-negligible impact on patients’ quality of life, waiting lists and healthcare costs. Therefore, future studies trying to identify the criteria that may better allow which patients to select for a second look TURB while avoiding an unnecessary intervention with possible risks and associated cost are needed to allow a personalized approach to even this one size fits all strategy.

ElectroMotive drug administration (EMDA) of Mitomycin C as first-line salvage therapy in high risk \u201cBCG failure\u201d non muscle invasive bladder cancer: 3\u2009years follow-up outcomes

BackgroundIn case of high grade non-muscle invasive bladder cancer (HG-NMIBC), intravesical BCG represents the first-line treatment; despite the “gold” standard therapy, up to 50% of patients relapse, needing radical cystectomy. Hence, alternative therapeutic strategies have been developed. The aim of the study was to evaluate a first-line salvage treatment with EMDA®-MMC in patients with HGNMIBC unresponsive to BCG.MethodsWe carried out a prospective, single-center, single-arm Phase II study in order to evaluate the efficacy (in terms of recurrence and progression) and the safety of the EMDA®-MMC treatment in 26 (21 male, 5 female) consecutive patients with “BCG refractory” HGNMIBC on a 3 years follow-up.EMDA®-MMC treatment consisted of 40 mg of MMC diluted in 100 ml of sterile water retained in the bladder for 30 min with 20 mA pulsed electric current.EMDA®-MMC regimen consisted of an induction course of 6 weekly instillations followed by a maintenance course of 6 monthly instillations.Follow-up was performed with systematic mapping biopsies of the bladder (with sampling in the prostatic urethra for men), voiding and washing urinary cytology, radiological study of the upper urinary tract.We performed Survival Kaplan-Meier curves and Log-rank test in order to analyze high grade disease-free survival.ResultsAt the end of follow-up, 16 patients (61.5%) preserved their native bladder; 10 patients (38.4%) underwent radical cystectomy, in 6 patients (23.1%) for recurrent HGNMIBC and in 4 patients (15.4%) for progression to muscle-invasive disease.At the end of follow-up, stratifying patients based on TNM classification (TaG3, T1G3, Cis, TaT1G3 + Cis), disease-free rates were 75, 71.4, 50 and 25%, respectively; survival curves showed statistically significant differences (p value < 0.05).Regarding toxicity, we reported severe adverse systemic event of hypersensitivity to the MMC in 3 patients (11.5%), and local side effects in 6 patients (26.1%).ConclusionsIn the field of alternative strategies to radical cystectomy, the EMDA®-MMC could be considered safe and effective in high-risk NMIBC unresponsive to BCG, as a “bladder sparing” therapy in selected patients. Multicenter studies with a larger number of patients and a longer follow-up might confirm our preliminary results.Trial registrationEudraCT2017-002585-43. 17 June 2017 (retrospectively registered).

Stem-Like Signature Predicting Disease Progression in Early Stage Bladder Cancer. The Role of E2F3 and SOX4.

The rapid development of the cancer stem cells (CSC) field, together with powerful genome-wide screening techniques, have provided the basis for the development of future alternative and reliable therapies aimed at targeting tumor-initiating cell populations. Urothelial bladder cancer stem cells (BCSCs) that were identified for the first time in 2009 are heterogenous and originate from multiple cell types; including urothelial stem cells and differentiated cell types—basal, intermediate stratum and umbrella cells Some studies hypothesize that BCSCs do not necessarily arise from normal stem cells but might derive from differentiated progenies following mutational insults and acquisition of tumorigenic properties. Conversely, there is data that normal bladder tissues can generate CSCs through mutations. Prognostic risk stratification by identification of predictive markers is of major importance in the management of urothelial cell carcinoma (UCC) patients. Several stem cell markers have been linked to recurrence or progression. The CD44v8-10 to standard CD44-ratio (total ratio of all CD44 alternative splicing isoforms) in urothelial cancer has been shown to be closely associated with tumor progression and aggressiveness. ALDH1, has also been reported to be associated with BCSCs and a worse prognosis in a large number of studies. UCC include low-grade and high-grade non-muscle invasive bladder cancer (NMIBC) and high-grade muscle invasive bladder cancer (MIBC). Important genetic defects characterize the distinct pathways in each one of the stages and probably grades. As an example, amplification of chromosome 6p22 is one of the most frequent changes seen in MIBC and might act as an early event in tumor progression. Interestingly, among NMIBC there is a much higher rate of amplification in high-grade NMIBC compared to low grade NMIBC. CDKAL1, E2F3 and SOX4 are highly expressed in patients with the chromosomal 6p22 amplification aside from other six well known genes (ID4, MBOAT1, LINC00340, PRL, and HDGFL1). Based on that, SOX4, E2F3 or 6q22.3 amplifications might represent potential targets in this tumor type. Focusing more in SOX4, it seems to exert its critical regulatory functions upstream of the Snail, Zeb, and Twist family of transcriptional inducers of EMT (epithelial–mesenchymal transition), but without directly affecting their expression as seen in several cell lines of the Cancer Cell Line Encyclopedia (CCLE) project. SOX4 gene expression correlates with advanced cancer stages and poor survival rate in bladder cancer, supporting a potential role as a regulator of the bladder CSC properties. SOX4 might serve as a biomarker of the aggressive phenotype, also underlying progression from NMIBC to MIBC. The amplicon in chromosome 6 contains SOX4 and E2F3 and is frequently found amplified in bladder cancer. These genes/amplicons might be a potential target for therapy. As an existing hypothesis is that chromatin deregulation through enhancers or super-enhancers might be the underlying mechanism responsible of this deregulation, a potential way to target these transcription factors could be through epigenetic modifiers.

Abstract 3163: Wnt7a promotes bladder cancer cell invasion via canonical Wnt/β-Catenin signaling

Abstract The mortality rate of urinary bladder cancer (UBC) increases dramatically, once low grade non-muscle invasive bladder cancer develops into muscle invasive bladder cancer. Hence, understanding the mechanisms underlying UBC invasion/metastasis and identification of biomarkers for invasion/metastasis will help clinicians to determine how to treat patients. Herein we generated two sublines from parental 5637 UBC cells with low invasive and high invasive capacities, which were designated as 5637 NMI and 5637 HMI, respectively. Mass spectrometry analysis was performed to identify the invasion-associated proteins. 5,987 proteins were quantified across all six samples with the coefficients among different channels all nearly 1.00, suggesting good biological repeatability and minor differences between 5637 HMI and NMI samples. Fold change &amp;gt; 1.2 and p value &amp;lt;0.05 were set as the cut-off standard, and 16 upregulated proteins and 26 downregulated proteins were identified in 5637 HMI cells. Among them, Wnt7a overexpression was confirmed in 5637 HMI cells compared to 5637 NMI cells, which was also associated with metastasis potential and worse clinical outcome in UBC patients. Depletion of Wnt7a in 5637 HMI and T24 cells reduced UBC cell invasion, with the reduction of active β-Catenin and its downstream target genes involved in epithelial-to-mesenchymal transition (EMT) and extracellular matrix (ECM) degradation. Consistently, introduction recombinant Wnt7a protein to 5637 NMI and J82 cells induced cell invasion, EMT, and the expression of ECM degradation-associated genes. Using luciferase reporter driven by the wildtype or mutants of matrix metalloproteinase 10 (MMP10) promoter, a canonical Wnt/β-Catenin regulation pattern was demonstrated. Four miRNAs downregulated in UBCs were predicted directly targeting Wnt7a using bioinformatics analysis. miRNA mimic could decrease Wnt7a expression, reduce the activation of MMP 10 reporter and suppress the invasion of 5637 HMI cells. Treatment of UBC cells with Wnt7a partially abrogated such miRNAs-induced invasion suppression. Taken together, our data identify a new miRNA/Wnt7a axis to mediate cell invasion and metastasis of UBC through canonical Wnt/β-Catenin pathway, which may offer prognostic and therapeutic opportunity. Citation Format: Ruimin Huang, Xiaojing Huang, Hongwen Zhu, Junlong Zhuang, Yu Dong, Hu Zhou, Hongqian Guo, Jun Yan. Wnt7a promotes bladder cancer cell invasion via canonical Wnt/β-Catenin signaling [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3163.

PO-488 AGR2 is a progression marker and a potential therapeutic target in non-muscle invasive bladder cancer

IntroductionBladder Cancer (BC) is the ninth most common cancer worldwide. Non-Muscle Invasive Bladder Cancer (NMIBC) is a heterogeneous disease that includes tumours with low and high risk of progression to muscle invasion. Nowadays, clinico-pathological features alone are not sufficient to effectively classify these patients and the discovery of new biomarkers is essential to increase quality of their life, who should only be treated with a radical high-morbidity approach, whenever needed, depending on the type of NMIBC tumour developed. ▵Np63 has previously been reported by our group as a protective factor of High Grade (HG) NMIBC progression: From the patients whose tumours showed nuclear ▵Np63 expression, none suffered disease progression after a median follow-up of 62,1 months. Furthermore, when ▵Np63 was knocked down (ΔNKO), cells showed higher proliferation and invasive capacities in vitro and higher tumour and metastasis formation in vivo. Interestingly, ΔNKO cells displayed AGR2 gene up-regulation and protein overexpression.Material and methodsTwo commercially available HG-NMIBC cell lines (RT112 and BF-TC905) were used to study the role of AGR2 in tumour progression. A stable knockdown of AGR2 (using shRNA technology) was performed on ΔNKO cells. In vitro functional studies, such as cell cycle, proliferation and invasion assays were performed comparing parental cells, ΔNKO cells and cells with knockdown of both ▵Np63 and AGR2 (AKO). The orthotopic mouse model is being used to study in vivotumour initiation and metastatic capacities. Immunofluorescence analyses of AGR2 in human HG-NMIBC tissue specimens will be performed to see if AGR2 expression is a prognostic and predictive biomarker for HG-NMIBC.Results and discussionsAfter knocking down AGR2 in RT112 and BF-TC905 ▵NKO cells, we observed a reversal of the aggressive phenotype of ΔNKO cells, with lower cell proliferation (cell cycle analysis and proliferation assay) and invasion, thus confirming the functional importance of AGR2. If in vivo results corroborate our hypothesis, we will start a collaboration with Dr. Li’s lab to test the efficiency of AGR2 antibody (Agtuzumab) intravesical therapy in NMIBC.ConclusionThis work uncovered AGR2 as a progression biomarker in HG-NMIBC, allowing a better understanding of the mechanisms of BC progression, and opening the possibility of novel therapies for patients affected by this disease, ultimately improving patients overall survival and quality of life.

MP83-19 MITOMYCIN C (MMC) PLUS BACILLUS CALMETTE E GUERÌN (BCG) SEQUENTIAL THERAPY FOR RECURRENT NON-MUSCLE INVASIVE BLADDER CANCER (NMIBC)

You have accessJournal of UrologyBladder Cancer: Non-invasive II1 Apr 2018MP83-19 MITOMYCIN C (MMC) PLUS BACILLUS CALMETTE E GUERÌN (BCG) SEQUENTIAL THERAPY FOR RECURRENT NON-MUSCLE INVASIVE BLADDER CANCER (NMIBC) Mauro Ragonese, Luca Di Gianfrancesco, Niccolò Lenci, Salvatore Marco Recupero, Giuseppe Palermo, and Marco Racioppi Mauro RagoneseMauro Ragonese More articles by this author , Luca Di GianfrancescoLuca Di Gianfrancesco More articles by this author , Niccolò LenciNiccolò Lenci More articles by this author , Salvatore Marco RecuperoSalvatore Marco Recupero More articles by this author , Giuseppe PalermoGiuseppe Palermo More articles by this author , and Marco RacioppiMarco Racioppi More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2018.02.2767AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES BCG therapy represents the gold standard treatment in high grade NMIBC to prevent recurrence and reduce the risk of progression. In patients who do not respond to BCG the gold standard is radical cystectomy. For patients not suitable or not willing to undergo radical surgery there are some bladder preservation strategies: a second BCG induction therapy and chemo-immunotherapy with BCG and MMC are among these options. METHODS We retrospectively evaluated data from 200 patients treated in our Department for a recurrent high grade NMIBC. All the patients were not suitable for surgery or they do not want to undergo radical surgery and were treated with different endovesical regimen. A group of 120 patients (Group 1) was treated with a schedule of combined therapy with 4 weekly instillations of MMC followed by 4 weekly instillations of BCG and a second group of 80 patients (Group 2) was treated with a second induction therapy of BCG. Patients with a complete response after induction were treated with maintenance therapy. We evaluated the complete response after the induction cycle and after a median follow up of 18 months and we compared the rate of side effects evaluated with the Common Terminology Criteria for Adverse Events (CTCAE) RESULTS Complete response after the first evaluation at three months was similar for the two groups, 67.5 % (81 patients) in group 1 and 63.7 % (51 patients) in group 2 without a statistically significant difference. At the median follow-up of 18 months a recurrent disease was present in 76 patients (38 %), 37 (46.2 %) from group 2 and 39 (37.5%) from group 1 (p < 0.01). 7 patients (6%) in group 1 and 6 patients (7.5 %) in group 2 had a progression to muscle-invasive disease. The regimen was well tolerated with low treatment toxicity, only Grade 1 and Grade 2 events were reported and the overall rate of complications was 25 % in group 1 and 11.2 % in group 2 (p < 0.01). No severe adverse events (Grade 3, 4 or 5) were reported and all the patients with a response after the induction complete the maintenance therapy. CONCLUSIONS Recurrent NMIBC is a great challenge for urologist in particular in patients not suitable for radical cystectomy. Sequential therapy and repeat BCG therapy are among these options and our preliminary findings indicate a better efficacy for the combination treatment. Further prospective randomized trials are needed to find out if this regimen could really become a valid alternative for these patients. © 2018FiguresReferencesRelatedDetails Volume 199Issue 4SApril 2018Page: e1123-e1124 Advertisement Copyright & Permissions© 2018MetricsAuthor Information Mauro Ragonese More articles by this author Luca Di Gianfrancesco More articles by this author Niccolò Lenci More articles by this author Salvatore Marco Recupero More articles by this author Giuseppe Palermo More articles by this author Marco Racioppi More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

MP08-07 THE IMPACT OF BLUE-LIGHT CYSTOSCOPY ON RESPONSE TO INDUCTION BCG IN PATIENTS WITH HIGH-GRADE NON-MUSCLE- INVASIVE BLADDER CANCER

You have accessJournal of UrologyBladder Cancer: Non-invasive I1 Apr 2018MP08-07 THE IMPACT OF BLUE-LIGHT CYSTOSCOPY ON RESPONSE TO INDUCTION BCG IN PATIENTS WITH HIGH-GRADE NON-MUSCLE- INVASIVE BLADDER CANCER Nina Mikkilineni, Samuel Antoine, Ashley Alford, Gen Li, Sven Wenske, G. Joel Decastro, James McKiernan, and Christopher Anderson Nina MikkilineniNina Mikkilineni More articles by this author , Samuel AntoineSamuel Antoine More articles by this author , Ashley AlfordAshley Alford More articles by this author , Gen LiGen Li More articles by this author , Sven WenskeSven Wenske More articles by this author , G. Joel DecastroG. Joel Decastro More articles by this author , James McKiernanJames McKiernan More articles by this author , and Christopher AndersonChristopher Anderson More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2018.02.312AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The use of blue light cystoscopy (BLC) during transurethral resection of bladder tumor (TURBT) is known to improve the detection of bladder cancer and reduce long-term recurrence rates. Intravesical Bacillus Calmette-Guerin (BCG) therapy is the standard of care for the treatment of high-grade non-muscle-invasive bladder cancer (NMIBC), and is associated with complete response rates of approximately 70%. We performed a retrospective study to investigate if response to induction BCG is affected by the use of BLC during pre-treatment TURBT in patients found with high-grade NMIBC. METHODS We retrospectively reviewed the charts of 148 patients with high-grade NMIBC at our institution who received induction BCG from 2014 to 2016. The primary exposure was white light cystoscopy (WLC) or BLC during the TURBT prior to induction BCG. Of the 148 patients, 103 patients had 3-month follow up with a cystoscopy with or without biopsy following completion of induction BCG. Disease recurrence was defined as visible or biopsy-proven tumor or positive urine cytology at 3 months. We used multivariate logistic regression to examine the association between type of cystoscopy and 3 month recurrence rate. Model covariates included stage, history of NMIBC, and prior intravesical therapy. RESULTS We identified 33 patients who had a BLC TURBT and 70 patients who had a WLC TURBT prior to induction BCG. Patients in the BLC group were more likely to have carcinoma in situ (67% vs. 47%, p=0.10), prior NMIBC (42% vs. 30%, p=0.31), and prior intravesical therapy (36% vs. 11%, p=0.007). At first cystoscopy after BCG induction, 13 patients (39%) in the BLC group and 24 (34%) in the WLC group had recurrent disease (p=0.78). On multivariate logistic regression, BLC did not have a lower recurrence rate compared to WLC (OR = 1.11, [95% CI 0.42, 2.93]; p=0.83). CONCLUSIONS There was no significant difference in 3 month recurrence rates after BCG induction for patients with BLC or WLC. The selection criteria for BLC differed from WLC, although our conclusion was unchanged after adjusting for confounding factors. BLC has been shown to improve detection of certain high risk tumors, but it is unclear if there is a therapeutic benefit to BLC for patients with known or suspected high risk disease. Further study is required to determine the value of BLC in improving response rates to intravesical BCG in patients with high grade NMIBC. © 2018FiguresReferencesRelatedDetails Volume 199Issue 4SApril 2018Page: e98 Advertisement Copyright & Permissions© 2018MetricsAuthor Information Nina Mikkilineni More articles by this author Samuel Antoine More articles by this author Ashley Alford More articles by this author Gen Li More articles by this author Sven Wenske More articles by this author G. Joel Decastro More articles by this author James McKiernan More articles by this author Christopher Anderson More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

P124 - Is repeated TURB always needed for a single high grade non-muscle invasive bladder cancer?

P124 - Is repeated TURB always needed for a single high grade non-muscle invasive bladder cancer?

Cytological and histological changes in the urothelium produced by electromotive drug administration (EMDA) and by the combination of intravescical hyperthermia and chemotherapy (thermochemotherapy)

The treatment of non muscle-invasive bladder cancer (NMIBC) encompasses a range of different procedures. Electromotive drug administration (EMDA) and chemo-hyperthermia (C-HT; Synergo) represent a minimally-invasive methods of intravesical instillation of therapeutic agents as mitomycin C (MMC). We selected patients with high grade NMIBC, BCG non responder, treated with EMDA/MMC and C-HT/MMC and we also examined the morphological changes in urine cytology samples.During the period from 2012 to 2014, 110 patients with high grade NMIBC, BCG refractory were selected. All cases examined were classified according to The Paris System Classification as negative for high urothelial carcinoma (NHGUC) or atypical urothelial cells (AUC) with a mean of follow-up of 15 months and the cytological diagnosis were confirmed by histological biopsies.In particular 50 patients were treated with EMDA/MMC and 60 patients underwent to C-HT/MMC. The morphological changes were evaluated in urine samples processed by Thin Prep method.In the 50 patients treated with EMDA/MMC, 35 samples were classified as NHGUC and 15 cases were classified as AUC, while in the 60 patients treated with C-HT/MMC, 43 samples were NHGUC and 17 cases were classified AUC.The increase of cellularity and nuclear size with the alteration of nuclear/cytoplasmatic ratio (N/C) were common in patients treated with EMDA/MMC and C-HT/MMC without clinical and histological evidence of recurrence of neoplasia. The hyperchromasia and irregular nuclear chromatin were rarely observed. The irregular nuclear membrane rarely identified in urine cytology after EMDA/MMC treatment, is a feature present in patients C-HT/MMC treated.

Effectivity of intravescical thermo-chemotherapy prophylaxis for patients with high recurrence and progression risk for non-muscle invasive bladder cancer.

Between February 2014 and December 2015, 18 patients with high risk NMIBC were enrolled. Patients were treated in an outpatient basis with 6 weekly induction sessions followed by monthly maintenance sessions with intravesical MMC in local hyperthermia with bladder wall thermo-chemotherapy (BWT) system (PelvixTT system, Elmedical Ltd., Hod Hasharon, Israel). The follow-up regimen included cystoscopy after the induction cycle and thereafter with regular intervals. Time to disease recurrence was defined as time from the first intravesical treatment to endoscopic or histological documentation of a new bladder tumour. Adverse events were recorded according to CTC 4.0 (Common Toxicity Criteria) score system. Mean age was 72 (32-87) years. 10 patients had multifocal disease, 9 had CIS, 6 had recurrent disease and 2 had highly recurrent disease (&gt; 3 recurrences in a 24 months period). 6 patients underwent previous intravesical chemotherapy with MMC. The average number of maintenance sessions per patient was 7.6. After a mean follow-up of 433 days, 15 patients (83.3%) were recurrence-free. 3 patients had tumour recurrence after a mean period of 248 days without progression. Side effects were limited to grade 1 in 2 patients and grade 2 in 1 patient. BWT seems to be feasible and safe in high grade NMIBC. More studies are needed to identify the subgroup of patients who may benefit more from this treatment.

Association between Metabolic Syndrome and Recurrence of Nonmuscle Invasive Bladder Cancer following bacillus Calmette-Guérin Treatment

IntroductionIntravesical bacillus Calmette-Guérin therapy is the gold standard adjuvant treatment for patients with high grade nonmuscle invasive bladder cancer. Despite the association between the metabolic syndrome and bladder cancer, the association between the metabolic syndrome and bacillus Calmette-Guérin failure is unknown. In this study we characterize disease recurrence following bacillus Calmette-Guérin in patients with and without metabolic syndrome. MethodsWe retrospectively evaluated the records of patients undergoing transurethral bladder tumor resection at our institution from 2012 to 2015 for nonmuscle invasive bladder cancer and identified those who received adjuvant bacillus Calmette-Guérin therapy. The metabolic syndrome was defined as having 3 of the 4 components of diabetes mellitus, hyperlipidemia, hypertension or body mass index 30 kg/m2or greater. The primary outcome was disease recurrence or progression. Descriptive statistics, chi-squared analysis, Kaplan-Meier survival analysis and Cox multivariable regression analyses were performed. ResultsHigh grade cancer was present in 83 of 90 (92.2%) patients. The metabolic syndrome was present in 27 of 90 (30%) patients. Median followup was 20 months. On Kaplan-Meier analysis patients with metabolic syndrome had worse disease-free survival than those without metabolic syndrome. On multivariable analysis body mass index 30 kg/m2 or greater was a significant predictor of recurrence or progression (HR 2.94, 95% CI 1.43–6.03). The presence of metabolic syndrome did not significantly affect the type of bacillus Calmette-Guérin failure. ConclusionsThe association between metabolic syndrome and failure to respond to bacillus Calmette-Guérin therapy is multifactorial but is in part associated with obesity. Elevated body mass index is strongly associated with recurrence or progression. Further studies are warranted to investigate the relationship between increased adiposity and response to bacillus Calmette-Guérin, especially as other novel immunotherapeutic agents are likely to enter the nonmuscle invasive bladder cancer space.

801 - ΔNp63 is critical for progression of high grade non-muscle invasive bladder cancer through deregulation of specific genetic pathways

801 - ΔNp63 is critical for progression of high grade non-muscle invasive bladder cancer through deregulation of specific genetic pathways

Phase 2 trial of dovitinib in Bacillus Calmette-Guerin (BCG) refractory urothelial carcinoma (UC) with tumor FGFR3 mutations or over-expression: Hoosier Cancer Research Network GU12-157.

4526Background: FGFR3 aberrations are common in low grade non-muscle invasive bladder cancer (NMIBC) tumors but decrease in frequency in metastatic UC. This multisite single-arm trial assessed the ...

Interobserver reproducibility of pathological variables in patients with high grade non-muscle invasive bladder cancer

The purpose of our study was to assess the interobserver variability in tumor stage and grade in patients with non-muscle invasive transitional cell bladder cancer. The prognostic relevance of grade and stage from both the initial and review diagnosis were determined. Pathological slides from 154 superficial bladder carcinomas were reviewed. Progression-free survival and hazard ratios for each stage and grade were calculated with Kaplan-Meyer method and Cox proportional model. There were significant interobserver differences in both the grading and staging of tumours. In pathology review overgrading was more frequent than undergrading, pathology review downstaged T category to stage Ta in 60,0 % of cases originally classified as stage T1.

Non-muscle invasive high grade urothelial carcinoma of the bladder. Which factors can influence understaging at the time of radical cystectomy?

To evaluate the main factors which influence understaging in patients with T1G3 non-muscle invasive bladder cancer (NMIBC). 109 patients with T1/G3 underwent transurethral resection of bladder tumor (TURBT) and then radical cystectomy (RC) with pelvic lymph nodes dissection. A number of variables were considered when evaluating the detection of understaging. We considered the patients age and gender, as well as the size, number, location and morphology of their tumor. We also considered coexistence of bladder carcinoma in situ (CIS), microscopic vascular invasion and deep lamina propria invasion. The level of experience of the surgeon was also analyzed. in RC samples muscle invasion, that is understaging, was detected in 74 (67.9%) patients, while 35 (32.1%) patients were appropriately staged. In these cohort of patients with high grade tumors, understaging was associated with deep lamina propria and microscopic vascular invasion, multiple tumors, tumor size &gt; 6 cm, tumor location (trigone and dome), presence of residual tumor; age, gender, tumor morphology, CIS associated, and experience of urological surgeon were not associated with clinical understaging. in our study, evaluating patients with high grade NMIBC at first TURBT, we identified some risk factors that need to be considered and that are able to increase the risk of understaging: deep lamina propria and microscopic vascular invasion, multiple tumors, tumor size &gt; 6 cm, tumor location (trigone and dome), presence of residual tumor. When these risk factors are present, performing an early cystectomy, and not a re-TURBT, could lower the risk of worse pathological finding due to rapid disease progression of the high grade tumors, and can prolong survival.

Quantitative Analysis of Differential Proteome Expression in Epithelial-to-Mesenchymal Transition of Bladder Epithelial Cells Using SILAC Method.

Epithelial-to-mesenchymal transition (EMT) is an essential biological process involved in embryonic development, cancer progression, and metastatic diseases. EMT has often been used as a model for elucidating the mechanisms that underlie bladder cancer progression. However, no study to date has addressed the quantitative global variation of proteins in EMT using normal and non-malignant bladder cells. We treated normal bladder epithelial HCV29 cells and low grade nonmuscle invasive bladder cancer KK47 cells with transforming growth factor-beta (TGF-β) to establish an EMT model, and studied non-treated and treated HCV29 and KK47 cells by the stable isotope labeling amino acids in cell culture (SILAC) method. Labeled proteins were analyzed by 2D ultrahigh-resolution liquid chromatography/LTQ Orbitrap mass spectrometry. Among a total of 2994 unique identified and annotated proteins in HCV29 and KK47 cells undergoing EMT, 48 and 56 proteins, respectively, were significantly upregulated, and 106 and 24 proteins were significantly downregulated. Gene ontology (GO) term analysis and pathways analysis indicated that the differentially regulated proteins were involved mainly in enhancement of DNA maintenance and inhibition of cell-cell adhesion. Proteomes were compared for bladder cell EMT vs. bladder cancer cells, revealing 16 proteins that displayed similar changes in the two situations. Studies are in progress to further characterize these 16 proteins and their biological functions in EMT.

Long-term oncological and functional outcomes of prostate sparing cystectomy: Single tertiary care institution experience.

413 Background: To analyze the oncological and functional outcome of Prostate Sparing Cystectomy (PSC) in the treatment of Bladder Cancer (BC) in a single tertiary care referral institution. Methods: At our institution, we treated 147 patients (1992 – 2014) with PSC for muscle invasive BC and recurrent high grade non-muscle invasive BC. Out of these, 64 patients received a standardized PSC technique featuring a Millin adenomectomy between 2001 – 2014 and composed the population of the study. Inclusion criteria were: Normal prostate examination and prostate specific antigen (PSA) +/- negative transrectal ultrasound guided biopsies; Frozen section negative for cancer in prostatic urethra prostate base, distal ureters and trigone. Oncological outcome was evaluated with Overall survival (OS); Cancer-specific survival (CSS); Recurrence-free survival (RFS). Functional outcomes: Continence - being pad-free or leak-free requiring ≤ 1 pad per day/night. Sexual function was assessed by self- reported erectile function sufficient for intercourse with or without medical treatment. Results: Median age was 62 years (IQR 55-67,5) with a median follow-up of 70 months (IQR 46-103). Thirty one (48,4%) patients ≤ pT1, 19 (29,7%) pT2, 14 (21,9%) ≥ pT3 at pathological diagnosis. Lymphadenectomy was performed in 95,3% of the cases. Positive lymph nodes were found in 4.9% patients and 3.1% patients showed positive surgical margins. Ten patients (15,7%) had Clavien grade 3–5 complications.The 5 and 10-y survival rates were OS: 84,7% and 68,3%; CSS: 84,7% and 77,5%; RFS: 76% and 65,5% respectively. No case of recurrence at the remnant prostatic urothelium was observed. Pad-free and leak-free continence, were achieved in 85.9% and 93.8% for daytime continence respectively, with 50% and 85,9% for night continence, respectively. Erectile function was maintained in 56,3%. Prostate cancer was diagnosed in 3 (4,7%) patients, 2 at the pathology report of the adenomectomy followed with active surveillance, and 1 was diagnosed during the follow up being treated with brachytherapy. Conclusions: PSC appears to be an oncologically safe procedure with adequate functional outcomes in treating selected patients with BC.

Global Identification and Differential Distribution Analysis of Glycans in Subcellular Fractions of Bladder Cells.

Compartmentalization of cellular components and their associated biological processes is crucial for cellular function. Protein glycosylation provides a basis for diversity of protein functions. Diversity of glycan composition in animal cells remains poorly understood. We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). An integrated strategy combining lectin microarray and mass spectrometry (MS) analysis was then applied to evaluate protein glycosylation of the four fractions. Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. Glycans in the latter category are considered potential markers for the corresponding organelles. The integrated strategy described here allows detailed examination of glycomes subcellular fraction with high resolution and sensitivity, and will be useful for elucidation of the functional roles of glycans and corresponding glycosylated proteins in distinct organelles.

Patients with High Grade Bladder Cancer, and Tumor-Free 3 and 6-Month Cystoscopy after bacillus Calmette-Guérin have a Good Prognosis without Further Maintenance bacillus Calmette-Guérin

IntroductionThe benefit of 3 years of maintenance bacillus Calmette-Guérin has been questioned. The outcome is reported for bacillus Calmette-Guérin treated patients who had negative 3 and 6-month cystoscopy results and were subsequently not treated with maintenance bacillus Calmette-Guérin. MethodsA retrospective, population based study of patients with high grade nonmuscle invasive bladder cancer treated with bacillus Calmette-Guérin was performed. Patients were included in analysis who had tumor-free cystoscopy findings 3 and 6 months after the start of treatment. No patient was treated with maintenance bacillus Calmette-Guérin after the 6-month cystoscopy. The Kaplan-Meier estimated 5-year survival rates were calculated. ResultsThree and 6-month cystoscopy revealed no tumor in 196 patients with a median age of 72 years. Of the patients 84% had carcinoma in situ or multiple and/or recurrent high grade tumors. Five-year recurrence-free, progression-free and disease specific survival was 69%, 95% and 98%, respectively. Median followup in 119 patients who were alive at the last contact was 79 months. Recurrence developed in 62 of 196 patients (32%), disease stage progressed to at least T2 in 13 (7%) and 7 (3%) died of bladder cancer. ConclusionsPatients without maintenance bacillus Calmette-Guérin after tumor-free 3 and 6-month cystoscopy have a low rate of progression and bladder cancer death. An alternative to 1 to 3 years of bacillus Calmette-Guérin maintenance may be no maintenance after the second tumor-free cystoscopy with re-treatment with bacillus Calmette-Guérin in case of recurrence.