Non-muscle Invasive Bladder Cancer Group Research Articles

Evaluation of Whole-Tumor Texture Analysis Based on MRI Diffusion Kurtosis and Biparametric VI-RADS Model for Staging and Grading Bladder Cancer.

to evaluate the feasibility of texture analysis (TA) based on diffusion kurtosis imaging (DKI) in staging and grading bladder cancer (BC) and to compare it with apparent diffusion coefficient (ADC) and biparametric vesical imaging reporting and data system (VI-RADS). In this retrospective study, 101 patients with pathologically confirmed BC underwent MRI with multiple-b values ranging from 0 to 2000 s/mm2. ADC- and DKI-derived parameters, including mean kurtosis (MK) and mean diffusivity (MD), were obtained. First-order texture histogram parameters of MK and MD, including the mean; 5th, 25th, 50th, 75th, and 90th percentiles; inhomogeneity; skewness: kurtosis; and entropy; were extracted. The VI-RADS score was evaluated based on the T2WI and DWI. The Mann-Whitney U-test was used to compare the texture parameters and ADC values between non-muscle-invasive bladder cancer (NMIBC) and muscle-invasive bladder cancer (MIBC), as well as between low and high grades. Receiver operating characteristic analysis was used to evaluate the diagnostic performance of each significant parameter and their combinations. The NMIBC and low-grade group had higher MDmean, MD5th, MD25th, MD50th, MD75th, MD90th, and ADC values than those of the MIBC and the high-grade group. The NMIBC and low-grade group yielded lower MKmean, MK25th, MK50th, MK75th, and MK90th than the MIBC and high-grade group. Among all histogram parameters, MD75th and MD90th yielded the highest AUC in differentiating MIBC from NMIBC (both AUCs were 0.87), while the AUC for ADC was 0.86. The MK75th and MK90th had the highest AUC (both 0.79) in differentiating low- from high-grade BC, while ADC had an AUC of 0.68. The AUC (0.92) of the combination of DKI histogram parameters (MD75th, MD90th, and MK90th) with biparametric VI-RADS in staging BC was higher than that of the biparametric VI-RADS (0.89). Texture-analysis-derived DKI is useful in evaluating both the staging and grading of bladder cancer; in addition, the histogram parameters of the DKI (MD75th, MD90th, and MK90th) can provide additional value to VI-RADS.

Serum metabolites as early detection markers of non-muscle invasive bladder cancer in Chinese patients.

Biomarkers of different stages and grades of bladder cancer (BC) are important in clinical work. The objective of our study was to investigate new biomarkers of early-stage BC with liquid chromatography-high resolution mass spectrometry (LC-HRMS) using serum samples. A total of 215 cases were included in our study, including 109 healthy adults as the control group and 106 non-muscle invasive bladder cancer (NMIBC) patients as the NMIBC group. Serum samples were collected from BC patients in the early stage, called NMIBC, and healthy people before surgery. We used LC-HRMS to distinguish the NMIBC group from the control group and the low-grade NMIBC group from the control group. An apparent difference between the NMIBC group and the control group was visualized by unsupervised principal component analysis (PCA). Metabolite panels for 16-hydroxy-10-oxohexadecanoic acid, PGF2a ethanolamide, sulfoglycolithocholate, and threoninyl-alanine were used to distinguish the two groups. The area under the curve (AUC) of the panels was 0.985, and the sensitivity and specificity were 98.63% and 98.59%, respectively. To distinguish the low-grade NMIBC group from the control group, serum metabolic profiling differences between the low-grade NMIBC group and control group samples were also analyzed. Metabolite panels of L-octanoylcarnitine, PGF2a ethanolamide, and threoninyl-alanine showed good discrimination performance. The AUC of the panels was 0.999, and the sensitivity and specificity were 97.8% and 100%, respectively. Metabolomics analysis of serum samples can distinguish the NMIBC group from the control group, particularly the early-stage low-grade NMIBC group.

Prediction of recurrence of non-muscle invasive bladder cancer: The role of androgen receptor and miRNA-2909.

The exact role of androgen receptor (AR) and miR-2909 in non-muscle invasive bladder cancer (NMIBC) remains controversial. Our aim is to assess the relationship between NMIBC recurrences with AR expression and miRNA-2909. This prospective controlled cohort study included 99 male patients with NMIBC (Ta-T1) who were treated by transurethral resection and 50 male patients as healthy control. We quantified blood AR messenger RNA variants 1 and 2 (AR1, AR2) and plasma miRNA-2909 with real-time quantitative polymerase chain reaction and the full length AR (AR-FL) using enzyme-linked immunosorbent assay in serum samples. In addition, AR1 and AR2 expression in tumor as well as normal tissues were analyzed. Kaplan-Meier survival curves and multivariate Cox analysis were used to identify independent predictors of recurrence-free survival. In comparison to control group, blood AR1, AR2, and serum AR-FL expression were significantly lower in the NMIBC group compared to plasma miRNA-2909 expression that was significantly higher in the control group. Blood AR1, AR2, and serum AR-FL were significantly correlated with higher tumor stage (pT1) while plasma miRNA-2909 was not. The median survival time (months) was significantly better for higher blood AR1 (34 vs. 21; P = 0.03), lower plasma miRNA-2909 (29 vs. 8; P <0.001), lower AR2 in normal tissues (32 vs. 22; P = 0.007). On multivariate analysis, serum free testosterone (hazards ratio [HR]: 8.9; 95% CI: 1.8-45.1; P = 0.008), serum AR1 (HR: 0.5; 95% CI: 0.3-0.9; P = 0.02), and plasma miRNA-2909 (HR: 5.8; 95% CI: 2.3-14.7; P = 0.0002), and tissue AR2 (HR: 2.6; 95% CI: 1.4-4.7; P = 0.002) were independent predictor for NMIBC recurrence. Blood and tissue levels of AR expression have a potential significant effect on NMIBC recurrence. Further studies are recommended to establish its exact role.

Evaluation of a fractional-order calculus diffusion model and bi-parametric VI-RADS for staging and grading bladder urothelial carcinoma.

To evaluate the feasibility of high b-value diffusion-weighted imaging (DWI) for distinguishing non-muscle-invasive bladder cancer (NMIBC) from muscle-invasive bladder cancer (MIBC) and low- from high-grade bladder urothelial carcinoma using a fractional-order calculus (FROC) model as well as a combination of FROC DWI andbi-parametric Vesical Imaging-Reporting and Data System (VI-RADS). Fifty-eight participants with bladder urothelial carcinoma were included in this IRB-approved prospective study. Diffusion-weighted images, acquired with 16 b-values (0-3600 s/mm2), were analyzed using the FROC model. Three FROC parameters, D, β, and μ, were used for delineating NMIBC from MIBC and fortumor grading. A receiver operating characteristic (ROC) analysis was performed based on the individual FROC parameters and their combinations, followed by comparisons with apparent diffusion coefficient (ADC) and bi-parametric VI-RADS based on T2-weighted images and DWI. D and μ were significantly lower in the MIBC group than in the NMIBC group (p = 0.001 for each), and D, β, and μ all exhibited significantly lower values in the high- than in the low-grade tumors (p ≤ 0.011). The combination of D, β, and μ produced the highest specificity (85%), accuracy (78%), and the area under the ROC curve (AUC, 0.782) for distinguishing NMIBC and MIBC, and the best sensitivity (89%), specificity (86%), accuracy (88%), and AUC (0.892) for tumor grading, all of which outperformed the ADC. The combination of FROC parameters with bi-parametric VI-RADS improved the AUC from 0.859 to 0.931. High b-value DWI with a FROC model is useful in distinguishing NMIBC from MIBC and grading bladder tumors. • Diffusion parameters derived from a FROC diffusion model may differentiate NMIBC from MIBC and low- from high-grade bladder urothelial carcinomas. • Under the condition of a moderate sample size, higher AUCs were achieved by the FROC parameters D (0.842) and μ (0.857) than ADC (0.804) for bladder tumor grading with p ≤ 0.046. • The combination of the three diffusion parameters from the FROC model can improve the specificity over ADC (85% versus 67%, p = 0.031) for distinguishing NMIBC and MIBC and enhance the performance of bi-parametric VI-RADS.

Microfluidic Assaying of Circulating Tumor Cells and Its Application in Risk Stratification of Urothelial Bladder Cancer.

Bladder cancer is characterized by its frequent recurrence and progression. Effective treatment strategies need to be based on an accurate risk stratification, in which muscle invasiveness and tumor grade represent the two most important factors. Traditional imaging techniques provide preliminary information about muscle invasiveness but are lacking in terms of accuracy. Although as the gold standard, pathological biopsy is only available after the surgery and cannot be performed longitudinally for long-term surveillance. In this work, we developed a microfluidic approach that interrogates circulating tumor cells (CTCs) in the peripheral blood of bladder cancer patients to reflect the risk stratification of the disease. In a cohort of 48 bladder cancer patients comprising 33 non-muscle invasive bladder cancer (NMIBC) cases and 15 muscle invasive bladder cancer (MIBC) cases, the CTC count was found to be considerably higher in the MIBC group compared with the NMIBC group (4.67 vs. 1.88 CTCs/3 mL, P=0.019), and was significantly higher in high-grade bladder cancer patients verses low-grade bladder cancer patients (3.69 vs. 1.18 CTCs/3mL, P=0.024). This microfluidic assay of CTCs is believed to be a promising complementary tool for the risk stratification of bladder cancer.

Transcriptome analysis of low-risk and high-risk non-muscular invasive bladder cancer patients to reveal disease progression related genes.

483 Background: According to the EAU prognostic risk classes, non-muscle invasive bladder cancer (NMIBC) is divided into low, medium, and high risk. Patients with high-risk NMIBC (T1/Tis, with high grade/G3, or CIS) represent a challenging group as they are at greater risk of recurrence and progression.Our specific aim was to investigate the biomarkers associated with progression and recurrence in NMIBC. Methods: Tumor tissue were collected from 70 patients with bladder cancer, including high-risk NMIBC (n = 44),low-risk NMIBC (n = 10) and (n = 16) MIBC. mRNA sequenced using the ABclonal Whole RNA-seq Lib Prep kit.The differentially expressed genes were identified using DESeq2 and the analysis of the association between genes expression and patient survival with NMIBC cohorts data(Jakob et al., 2016). RNAseq data of human bladder cancer cell lines were achieved from the Cell Model Passports website (https://cellmodelpassports.sanger.ac.uk/). Results:A total of 456 genes are significantly high expressed in high-risk NMIBC group compared with the low-risk NMIBC group. Combined with MIBC expression data,we found 16 genes with consistently increasing expression from the low-risk NMIBC group to high-risk NMIBC group and MIBC group using a fold change of at least 2, and a false discovery rate (FDR) of 0.05.Among these genes,14 genes were also highly expressed in human bladder cancer cell lines. Survival analysis by Kaplan-Meier,we finally identified 13 high-expressed genes, including KRT6A、SPHK1、S100A9、SLC16A1、CDC25B、NELL2、PREX1、C15orf48、AKR1B10、CERCAM、PKMYT1、UCHL1、SLC16A1, were significantly associated with poor progression-free survival(p&lt;0.05). Conclusions: We identified a set of 13 genes that may predict the progression of NMIBC and may serve as molecular targets for NMIBC therapy. these study results need to be confirmed in larger research studies.

Do urinary mast cell mediators predict immune response to BCG in patients with primary high-grade non-muscle invasive bladder cancer?

Mast cells play a critical role in cancer-associated immunity. We aimed to determine the predictive value of urinary mast cell mediators in patients with non-muscle invasive bladder cancer (NMIBC) treated with Bacillus Calmette-Guérin (BCG) immunotherapy. In this prospective study, 19 patients who received immunotherapy because of NMIBC (Group 1) and 19 healthy participants (Group 2) were enrolled. Urine samples were collected to assay N-methylhistamine, histamine, and tryptase levels immediately before the first BCG instillation, immediately after the third and sixth instillations, and 4 weeks after the sixth instillation in Group 1 and at a single visit in Group 2. The changes in urinary markers because of BCC response, BCG instillation, and the presence of NMIBC were assessed. The average age was 56.1±10.5years in Group 1 and 52.6±9.7years in Group 2. Fourteen patients had high-grade Ta tumours and five had T1 tumours. While 12 patients had responded to the BCG, seven patients did not respond to the BCG. There was no correlation between mast cell mediators and BCG response. The N-methylhistamine and histamine levels significantly increased with the onset of immunotherapy, and N-methylhistamine levels significantly decreased when immunotherapy was terminated (P<.05). The pre-BCG estimated marginal mean values of N-methylhistamine were significantly higher in Group 1 than in Group 2 (P<.05). Our study is the first to identify the changes in mast cell mediators with the onset of immunotherapy and in presence of bladder cancer. However, these mediators cannot predict patients' response to immunotherapy.

Effects of P-MAPA as Immunomodulator on Markers for Energy Metabolism in Bladder Cancer

Introduction This study characterized and compared cellular energetic metabolism features in the treatment of chemically induced non-muscle invasive bladder cancer (NMIBC) in Fischer 344 rats that were submitted to intravesical immunotherapies with P-MAPA (Protein aggregate magnesium-ammonium phospholinoleate-palmitoleate anhydride) and Bacillus Calmette-Guérin (BCG). Methods Rat urinary bladder samples from Control, NMIBC (Cancer), NMIBC+BCG and NMIBC+P-MAPA groups were submitted to histopathological and western blotting analyses for the following proteins: GLUT 1, PFK, GAPDH, HADHSC, β-F1-ATPase, AMPK and mTOR. Results P-MAPA Intravesical treatment was effective in tumor regression and histologic recovery of the urinary bladder in the NMIBC. There was a significant increase in protein levels of GLUT 1, mTOR, GAPDH and HADHSC in the NMIBC+P-MAPA group. It was observed an increase in protein levels of PFK and β-F1-ATPase and a significant reduction in protein levels of AMPk in the NMIBC group. Conclusions Our results showed that immunotherapy with P-MAPA may be an alternative in the treatment of NMIBC, especially in cases where BCG therapy failure, as evidenced by the effect of P-MAPA on tumor regression.

MP73-05 BCG-UNRESPONSIVE NMIBC, COMPARISON OF 2 ND -LINE INTRAVESICAL INSTILLATION THERAPY

You have accessJournal of UrologyBladder Cancer: Non-invasive IV (MP73)1 Apr 2020MP73-05 BCG-UNRESPONSIVE NMIBC, COMPARISON OF 2ND-LINE INTRAVESICAL INSTILLATION THERAPY Gal Rinott Mizrahi*, Yuval Freifeld, Ido Orlin, Ilona Pilosov, Boris Friedman, and Yoram Dekel Gal Rinott Mizrahi*Gal Rinott Mizrahi* More articles by this author , Yuval FreifeldYuval Freifeld More articles by this author , Ido OrlinIdo Orlin More articles by this author , Ilona PilosovIlona Pilosov More articles by this author , Boris FriedmanBoris Friedman More articles by this author , and Yoram DekelYoram Dekel More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000000959.05AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Intravesical instillation of live attenuated bacillus Calmette-Guérin (BCG) is the gold standard treatment for intermediate and high-risk Non-Muscle-Invasive Bladder Cancer (NMIBC), with proven effects on reducing recurrence, progression, and death. However, it is not effective in all patients, and recurrence after adequate BCG therapy can lead to progression and to a life-threatening disease. For this reason, the current recommendation for BCG-unresponsive disease is early radical cystectomy. In practice, most patients at that point prefer less aggressive treatment or are un-fit for radical surgery and therefore are treated with 2nd-line intravesical instillations. The objective of the current study is to compare 2nd line treatments for BCG-unresponsive disease. METHODS: This is a retrospective study based on a single institutional database. The study population included patients with BCG-unresponsive disease treated in our department during 2008-2016. BCG-unresponsive disease was defined as persistence or progression within 12 months of adequate BCG therapy. For each patient data regarding 2nd line therapy and oncologic outcome was obtained. RESULTS: Of 315 patients treated with BCG during the defined period, 68 patients had BCG-unresponsive disease. Presentation of unresponsive disease was with muscle invasive/metastatic disease in 25% of patients and with NMIBC in 75% of patients (n=51). In the NMIBC group 6 patients had early cystectomy, 21 were treated with re-induction BCG and 23 were treated with Thermo-chemotherapy Mitomycin C (SYNERGO). Response to 2nd line treatment was observed in 33.3% and 39.1% in the BCG and SYNERGO groups respectively. Immediate progression to muscle invasive/metastatic disease was observed in 19% and 26.1% in the BCG and SYNERGO groups respectively. In each group 3 patients further progressed to metastatic disease during follow-up. CONCLUSIONS: We found a response rate to 2nd line intravesical instillation therapy in BCG-unresponsive disease of 35% with no significant difference between re-induction with BCG or SYNERGO. Progression to muscle invasive/metastatic disease was observed in 35% of patients with no significant difference between groups. These results emphasize the need to present to patients with BCG-unresponsive disease, the poor prognosis of their disease and encourage strongly towards early cystectomy. For patient who refuse or are un-fit, there is no significant differences between re-induction with BCG or SYNERGO. Source of Funding: non © 2020 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 203Issue Supplement 4April 2020Page: e1122-e1123 Advertisement Copyright & Permissions© 2020 by American Urological Association Education and Research, Inc.MetricsAuthor Information Gal Rinott Mizrahi* More articles by this author Yuval Freifeld More articles by this author Ido Orlin More articles by this author Ilona Pilosov More articles by this author Boris Friedman More articles by this author Yoram Dekel More articles by this author Expand All Advertisement PDF downloadLoading ...

MP61-13 DO URINARY-BASED MOLECULAR BIOMARKER HAVE THE ABILITY TO PREDICT THE GRADE OF NON-MUSCLE INVASIVE BLADDER CANCER GRADE?

INTRODUCTION AND OBJECTIVE: Micro-RNA155 (mir-155), MicroRNA-200b (mir-200b), human telomerase reverse transcriptase (hTERT), and E2F3 transcription factor had a role in bladder cancer pathogenesis. We aim to assess their role in diagnosis the non-muscle invasive bladder cancer (NMIBC) grade compared to histopathological results. METHODS: A registered prospective trial (NCT03591367) for 64 NMIBC patients. From voided urine samples, total RNA extract from sedimented urothelial cells were analysed by a reverse transcriptase-polymerase chain reaction assay for the presence of mir-155, mir-200b, hTERT and E2F3 transcript. A receiver operating characteristics (ROC) was plotted in order to choose the best cut-off point. Sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were calculated for each test. RESULTS: Patients demographic were comparable (table 1). The expression levels of the mir-155 didn’t show difference between high grade (HG) and low grade (LG) (p=0.06) (table 1). Both mir-155 and mir-200b had the lowest AUC (table 2). E2F3 and hTERT expression in urine was higher in HG NMIBC group than in LG NMIBC (P < 0.001) (table 1). The sensitivity, specificity, accuracy of E2F3 were 86%, 76.2%, and 82.8%, respectively. The sensitivity, specificity, accuracy of hTERT were 86%, 66.7%, and 79.7%, respectively (table 2 and figure 1). CONCLUSIONS: As molecular urinary biomarkers; E2F3 and hTERT have the highest potential for prediction of the grade of NMIBC to either low or high grade.Source of Funding: No

Gene mutation detection of urinary sediment cells for NMIBC early diagnose and prediction of NMIBC relapse after surgery.

Early diagnose of bladder cancer could lead to good prognosis and high 5-year-survival rate. Among bladder cancer, about 75% patients with were nonmuscle-invasive bladder cancer (NMIBC). Patients were painful and easily get infected during bladder cancer diagnosis, which mainly depends on invasive cystoscopy and low-sensitivity urine exfoliation cytology. Meanwhile, relapse after surgery was also becoming the major problem for patients. Exploring noninvasive, high-sensitivity, and painless method is very important and meaningful for NMIBC treatment.Firstly, we found potential related gene mutation sites for NMIBC by searching COSMIC database and related study. Urinary sediment cells of patients both in normal group (patients with benign) and NMIMC group were collected before and after operation for potential gene mutation site detecting. Meanwhile, the urinary sediment cells of relapse patients and good prognosis people in NMIBC group after surgery were also collected for further Gene mutation detection and NMIBC relapse after surgery prediction.Fourteen genes (152 mutation sites) were selected between 95 NMIBC patients and 67 control patients, which were FGFR3, TP53, PIK3CA, and others. Compared with control group, mutation ratio of above 14 genes was higher in NMIBC group. NMIBC diagnose model was established by 5 times cross-validation and had a good effects, which included the all mutation site in FGFR3, TP53, PIK3CA, ARID1A, STAG2, and KTM2D. On the contrary, the relapse rate was 30.5% among 95 patients for about 1.5-year follow-up time. Compared with control group, smoking rate and tumor grade were higher in relapse group. Meanwhile, mutation rate of FGFR3, TP53, PIK3CA, ERBB3, and TSC1 in relapse group were higher than that in normal group. According to the mutation sites of FGFR3, TP53, PIK3CA, and ERBB3 and the combination of urinary sediment cells genetic mutation and relapse status, a predicted model for NMIBC relapse was also established, which had 90% accuracy.The diagnosed NMIBC model (based on FGFR3, TP53, PIK3CA, ARID1A, STAG2, and KTM2D gene mutation) and predicted relapse model (based on FGFR3, TP53, PIK3CA, and ERBB3 gene mutation) possess high accuracy and would be applied in early diagnose and early predicting relapse of patients.

Role of OncoTherad immunotherapy in the regulation of toll-like receptors-mediated immune system and RANK/RANKL signaling: New therapeutic perspective for non-muscle invasive bladder cancer.

e16004 Background: The new modalities for treating patients with non-muscle invasive bladder cancer (NMIBC) for whom Bacillus Calmette-Guerin (BCG) has failed or is contraindicated are recently increasing due to the development of new drugs. In this scenario, a new perspective is represented by OncoTherad immunomodulator. OncoTherad is a nanostructured inorganic phosphate complex associated to glycosidic protein, developed by University of Campinas/ Brazil, which exhibits antitumor properties. This study detailed and characterized the therapeutic effects of OncoTherad based on activation of Toll-Like Receptors (TLRs) signaling pathways and regulation of receptor activator of nuclear factor kβ (RANK)/RANK ligand (RANKL) cytokine system in an animal model of NMIBC, as well as, compared these effects with BCG treatment. Methods: Fisher 344 female rats were submitted to NMIBC induction with N-methyl-N-nitrosourea (MNU). MNU treated animals were further divided into 3 groups (10 animals per group): the NMIBC group received 0.30 mL of saline solution; the NMIBC-BCG group received of 2 mg/mL of BCG; the NMIBC-OncoTherad group received 20 mg/Kg dose of OncoTherad. All animals were treated intravesically every other week for 6 weeks. Results: Our results demonstrated that OncoTherad intravesical immunotherapy led to distinct activation of TLRs 2 and 4-mediated innate immune system, resulting in increased interferons signaling pathway, which was more effective (80.0%) in the NMIBC treatment, when compared to BCG treatment (20.0%). Interferon signaling pathway activation induced by OncoTherad led to increase of iNOS expression, resulting in apoptosis and histopathological recovery. Also, OncoTherad immunotherapy decreased RANK/RANKL expression, resulting in reduced regulatory T (Treg) cells. Conclusions: The decreased of RANK/RANKL expression by OncoTherad was fundamental to up-regulation interferon signaling pathway and in suppresion of abnormal cell proliferation. Thus, OncoTherad immunotherapy could be considered an important therapeutic strategy for NMIBC, as well as, opens a new perspective for treatment of BCG-refractory or relapsed patients.

Upregulation of FGFR3 and HIF-1α expression in muscle invasive bladder cancer

BACKGROUND The major risks in patients diagnosed with non-muscle invasive bladder cancer (NMIBC)are recurrence, progression of muscle invasive bladder cancer (MIBC), and metastasis. Biological markers such as fibroblast growth factor receptor-3 (FGFR3) and hypoxia-inducible factor-1α (HIF-1α) are related to muscle invasiveness of bladder cancer. This study was aimed to analyze the expression of FGFR3 and HIF-1α to predict muscle invasiveness in bladder cancer patients.METHODS This was an observational study with a case-control design. Sixty patients with bladder cancer, who underwent histopathology examinations at the Department of Pathology, Faculty of Medicine, Universitas Sumatera Utara/H. Adam Malik Hospital from January 2012 to December 2015, were included in this study. Samples were then classified into 30 NMIBC and 30 MIBC groups. All samples were analyzed with an immunohistochemistry assay for FGFR3 and HIF-1α. H-scores were used to determine the relationships between each group.RESULTS FGFR3 was expressed in 29 (96.7%) patients of the NMIBC group, and 23 (76.7%) patients of the MIBC group (p=0.026, OR=8.8; 95% CI=1.01–76.96). HIF-1α was expressed in only 1 (3.33%) patient of the NMIBC group, and 15 (50%) patients of the MIBC group (p&lt;0.001, OR=29; 95% CI=3.49–241.13).CONCLUSIONS There was a difference in upregulation of FGFR3 and HIF-1α expression in both the NMIBC and MIBC groups.

A novel messenger RNA and long noncoding RNA signature associated with the progression of nonmuscle invasive bladder cancer.

To explore the molecular mechanism of nonmuscle invasive bladder cancer (NMIBC), matched normal, and cancer tissues of 10 NMIBC were examined for RNA sequencing. We profiled the messenger RNA (mRNA) and long noncoding RNA (lncRNA) expression of patients with NMIBC. Differentially expressed mRNAs and lncRNAs were screened between cancer and normal tissues and validated by quantitative polymerase chain reaction (qPCR), and lncRNA-mRNA-miRNA interaction network was constructed. A total of 91 upregulated and 190 downregulated genes and 34 upregulated and 58 downregulated lncRNAs were screened from the sequencing result. The differentially expressed mRNAs were enriched in focal adhesion, rap1 signaling pathway, Hippo signaling pathway, PI3K-Akt signaling pathway, extracellular matrix (ECM)-receptor interaction, Ras signaling pathway, and mitogen-activated protein kinases signaling pathway, of which some pathways were involved in the cancer development. In the RNA sequencing, KIT and laminin subunitγ γ3 (LAMC3) were significantly downregulated in the NMIBC group compared with the normal group. The results of quantitative reverse transcription PCR showed that the expression of LAMC3 and KIT were significantly decreased in the NMIBC group compared with the normal group. The lncRNA-mRNA-miRNA interaction network was constructed by Cytoscape software to further investigate the interaction correlations. The results implied that KIT and LAMC3 might regulate the lncRNAs (such as ENST00000445707, ENST00000501122, ENST00000505254, ENST00000528986, ENST00000557661, ENST00000602964, ENST00000614517, ENST00000620864, and ENST00000623414) by the miRNAs (such as hsa-let-7f-2-3p, hsa-miR-125a-3p, hsa-miR-134-3p, hsa-miR-191-5p, hsa-miR-210-5p, hsa-miR-30a-5p, hsa-miR-30d-5p, hsa-miR-30e-5p, hsa-miR-92a-2-5p, and hsa-miR-95-3p), and finally played a role in the development of NMIBC cancer. Altogether, our study preliminarily indicated that KIT and LAMC3 might play a crucial role in the development of NMIBC cancer via a complex mRNA-lncRNA-miRNA regulatory network.

Bladder Cancer-Specific Nuclear Matrix Proteins-4 May Be a Potential Biomarker for Non-Muscle-Invasive Bladder Cancer Detection.

Aims Bladder cancer–specific nuclear matrix protein-4 (BLCA-4) is a protein expressed mainly in bladder cancer tissues. Therefore, the aim of this study was to investigate its assisting diagnostic potential in non-muscle-invasive bladder cancer (NMIBC). Methods Twenty patients with NMIBC, 20 with benign prostatic hyperplasia (BPH), and 20 normal controls were included in this study. Blood and urine samples were collected from all patients. Moreover, cancer foci and adjacent tissue samples were collected from NMIBC patients, and normal bladder tissue samples were collected from patients with BPH. A competitive enzyme-linked immunosorbent assay was used to determine the BLCA-4 level in serum and urine, and immunohistochemistry was used to examine BLCA-4 expression in bladder cancer, adjacent, and normal tissues. Results Median urinary BLCA-4 levels in the NMIBC, BPH, and normal control groups were 0.759 ng/mL, 0.309 ng/mL, and 0.171 ng/mL, respectively. Urinary BLCA-4 level was significantly higher in the NMIBC group than in the other 2 groups (P < 0.01); meanwhile, the BPH group was higher than the normal control group (P < 0.05). Median serum BLCA-4 levels in the NMIBC, BPH, and normal control groups were 5.680 ng/mL, 5.928 ng/mL, and 5.473 ng/mL, respectively, showing no significant difference among groups (P > 0.05). Conclusion As a new marker of bladder cancer, urinary BLCA-4 level detection might apply for clinical diagnosis or postoperative monitoring for NMIBC.

Exploring the roles of urinary HAI-1, EpCAM & EGFR in bladder cancer prognosis & risk stratification.

ObjectivesTo investigate whether elevated urinary HAI-1, EpCAM and EGFR are independent prognostic biomarkers within non-muscle-invasive bladder cancer (NMIBC) patients, and have utility for risk stratification to facilitate treatment decisions.ResultsAfter accounting for EAU risk group in NMIBC patients, the risk of BC-specific death was 2.14 times higher (95% CI: 1.08 to 4.24) if HAI-1 was elevated and 2.04 times higher (95% CI: 1.02 to 4.07) if EpCAM was elevated. The majority of events occurred in the high-risk NMIBC group and this is where the biggest difference is seen in the survival curves when plotted for EAU risk groups separately. In MIBC patients, being elevated for any of the three biomarkers was significantly associated with BC-specific mortality after accounting for other risk factors, HR = 4.30 (95% CI: 1.85 to 10.03).Patients and MethodsUrinary levels of HAI-1, EpCAM and EGFR were measured by ELISA in 683 and 175 patients with newly-diagnosed NMIBC and MIBC, respectively, recruited to the Bladder Cancer Prognosis Programme. Associations between biomarkers and progression, BC-specific mortality and all-cause mortality were evaluated using univariable and multivariable Cox regression models, adjusted for European Association of Urology (EAU) NMIBC risk groups. The upper 25% of values for each biomarker within NMIBC patients were considered as elevated. Exploratory analyses in urine from MIBC patients were also undertaken.ConclusionUrinary HAI-1 and EpCAM are prognostic biomarkers for NMIBC patients. These biomarkers have potential to guide treatment decisions for high-risk NMIBC patients. Further analyses are required to define the roles of HAI-1, EpCAM and EGFR in MIBC patients.

Outcomes of subsequent non-muscle-invasive bladder cancer treated with intravesical Bacillus Calmette-Guérin after radical nephroureterectomy for upper urinary tract urothelial carcinoma.

To describe the clinicopathological characteristics and prognosis of subsequent non-muscle-invasive bladder cancer (NMIBC) after radical nephroureterectomy (RNU) for upper urinary tract urothelial carcinoma (UTUC), and particularly its response to intravesical Bacillus Calmette-Guérin (BCG). An observational study was conducted in 1463 patients with UTUC who had undergone RNU and in 1555 patients with primary NMIBC. Of the 1463 patients with UTUC, 256 (17%) subsequently developed NMIBC (UTUC-NMIBC group) and were available for the analysis. The clinicopathological background and outcomes, including intravesical recurrence-free survival and bladder progression-free survival, were compared between the patients with UTUC-NMIBC and the patients with primary NMIBC treated with intravesical BCG. Propensity score matching was performed to adjust for the potential differences in the backgrounds of the two groups. To validate the utility of the CUETO scoring model in the UTUC-NMIBC group, risk scores were calculated and compared with the published probabilities for recurrence and progression. Compared with the unadjusted primary NMIBC group (n = 352), the UTUC-NMIBC group (n = 75) were found to have a worse prognosis for intravesical recurrence and progression, before propensity score matching. After propensity score matching for potential confounding factors, however, a worse prognosis was observed only for intravesical recurrence. The validation test of the CUETO scoring model for the UTUC-NMIBC group showed a significant difference in the rate of intravesical recurrence and progression for the 0-4 and 5-6 score groups between the UTUC-NMIBC group and the CUETO risk table reference data. Compared with the primary NMIBC group, the UTUC-NMIBC group had a worse prognosis after intravesical BCG, especially with regard to intravesical recurrence. This suggests that patients with UTUC-NMIBC are inherently poor responders to BCG exposure. An optimal treatment strategy and risk scoring model to select patients for adjuvant intravesical BCG, chemotherapy or immediate radical cystectomy should be established.

Molecular Assessment of Non-Muscle Invasive and Muscle Invasive Bladder Tumors: Mapping of Putative Urothelial Stem Cells and Toll-Like Receptors (TLR) Signaling

Purpose: The main objectives of this study were to characterize and compare the urothelial stem cells (healthy and cancer cells) and TLRs features in the urinary bladder of men without lesionsand with non-muscle-invasive and muscle invasive urothelial tumors. Materials and Methods: Thirty samples of the urinary bladder of 50 to 80-year-old men, with and without diagnosis of malignant urothelial lesions were used. The 30 samples were divided into 3 groups (n = 10 per group): Normal Group; Non-Muscle Invasive Bladder Cancer Group; Muscle Invasive Bladder Cancer Group. The samples were histopathologically and immunohistochemically analyzed. The study was conducted at teaching Hospital of the University of Campinas (UNICAMP). Results: The CD44 and CD133 immunoreactivities were significantly intense in the muscle-invasive cancer group when compared to the other groups. The ABCG2 biomarker demonstrated intense immunoreactivities in both non-muscle and muscle invasive groups, and absent immunoreactivity in the normal group. All groups showed weak CD117 immunoreactivity. Putative Healthy Stem Cells (CD44/CD133/ CD117+) occurred in all groups. Putative Cancer Stem Cells (CD44/CD133/ABCG2+) only occurred in the non-muscle and muscle invasive cancer groups. TLR2 immunoreactivity was significantly lower in the non-muscle invasive cancer group and absent in the muscle invasive cancer group. TLR4 immunoreactivity was significantly lower in both cancer groups. Conclusions: This study leads us to the conclusion that putative cancer stem cell occurrence was sensitive to the decreased in TLR2 and TLR4 immunoreactivities. Also, TLR2 and TLR4 demonstrated their involvement in the regulation of the different biomarkers for putative healthy and cancer urothelial stem cells, probably acting as negative regulators of urothelial carcinogenesis. Taken together data obtained suggest that use of TLRs agonists could be a promising alternative for the treatment of non-muscle and muscle invasive bladder tumors.

Bladder cancer invasion predictability based on preoperative neutrophil–lymphocyte ratio

An increased pretreatment neutrophil-lymphocyte ratio (NLR) is associated with poor prognosis in colorectal, gastric, and ovarian cancer; malignant mesothelioma; and renal cell carcinoma. The present study aims to define the predictive value of preoperative peripheral blood count NLR in non-muscle-invasive bladder cancer (NMIBC) and muscle-invasive disease (MIBC) patients. There were in total 291 patients, 241 males and 50 females. Out of these, 156 male and 36 female patients were in the NMIBC group and 85 male and 14 female patients in the MIBC group. In the NMIBC group, 172 patients had low-grade and 20 high-grade papillary urothelial carcinoma. The mean age of the patients in the NMIBC group was 64 ± 13, ranging from 27 to 97. The mean age of the patients in MIBC group was 70.5 ± 10, ranging from 27 to 95. A statistically significant relation between patient ages and tumor invasiveness was determined (p = 0.023, 95 % confidence interval (CI) 63.3-66.7). The mean tumor size of the NMIBC group was 2.1 ± 1.09 (cm) (range 0.5-8), and of MIBC group 3.6 ± 1.5 (cm) (range 0.8-9). There was a statistically significant relation between the tumor size and invasiveness (p = 0.002, 95 % CI 2.8-4.4). In the NIMBC group, 149 (77.6 %) of them have NLR ≤ 2.5 and 43 (22.4 %) have NLR > 2.5. Also, in MIBC, 67 (67.7 %) of them have NLR ≤ 2.5 and 32 (32.3 %) have NLR > 2.5. The mean NLR in the NMIBC group was 2.4 ± 0.1 (range 0.08-6.49, 95 % CI 1.52-2.71) and in the MIBC 2.9 ± 0.2 (range 0.08-16.72, 95 % CI 1.67-2.97). In terms of NLR, there was a statistically significant difference between the NMIBC and MIBC groups (p = 0.028). Platelet-lymphocyte ratio (PLR) of the two groups was also analyzed. The PLR of the NMIBC group was 12.8 ± 15.1 (range 3.38-19.1) and of the MIBC 13.6 ± 8.78 (range 0.18-63), yet there was not any statistically significant difference (p = 0.810, 95 % CI 11.4-14.8) (Table 1). The correlation tests revealed a positive correlation between the age (r = 0.144, p = 0.024), tumor size (r = 0.193, p = 0.02), and tumor invasiveness NLR (r = 0.138, p = 0.031). NLR can be used to determine tumor invasiveness as a cost-effective, common, and simple biomarker in bladder cancer (BC).

Combined Chemohyperthermia: 10-Year Single Center Experience in 160 Patients with Nonmuscle Invasive Bladder Cancer

Nonmuscle invasive bladder cancer is characterized by a high recurrence rate. New adjuvant treatments are needed to decrease this high number ofrecurrences. We present the results of more than 10 years of experience with chemohyperthermia in patients with nonmuscle invasive bladder cancer. Using standardized medical record forms we prospectively collected patient and tumor characteristics of patients treated with chemohyperthermia between 2002 and 2013. Median followup was 75.6 months. Recurrence-free survival was the primary objective. The secondary objective was to observe recurrence-free survival differences in 1) the epirubicin group vs the mitomycin group and 2) the highly recurrent (greater than 2 recurrences in 24 months) nonmuscle invasive bladder cancer group vs the other groups. A total of 160 patients with nonmuscle invasive bladder cancer were included in study, including 20 (13%) treated with epirubicin and 129 (81%) previously treated with bacillus Calmette-Guérin. One and 2-year recurrence-free survival was 60% and 47%, respectively. Muscle invasive progression was seen in 4% of cases. Two-year recurrence-free survival in the epirubicin and mitomycin groups was 55% and 46%, respectively (p = 0.30). The highly recurrent nonmuscle invasive bladder cancer group had significant decreased recurrence-free survival compared to other groups (p <0.01). Patients treated with 2 or fewer vs greater than 2 transurethral bladder tumor resections before chemohyperthermia had higher recurrence-free survival (p = 0.01). On multivariable analysis the highly recurrent cancer criteria remained independently associated with decreased recurrence-free survival (HR 2.40, 95% CI 1.30-4.43, p = 0.01). Chemohyperthermia is an effective approach to nonmuscle invasive bladder cancer for which standard intravesical treatments fail. Patients with highly recurrent disease before chemohyperthermia have lower recurrence-free survival. Furthermore, recurrence-free survival appears to improve with earlier chemohyperthermia. No significant differences were observed between the 2 chemotherapy agents.