Faouzia Ajili

Prognostic Value of Tumor-associated Macrophages Count in Human Non-muscle-invasive Bladder Cancer Treated by BCG Immunotherapy

Purpose: Tumor-associated macrophages can regulate the growth of various cancers positively or negatively. Intravesical bacillus Calmette-Guerin instillation is now the gold standard treatment for bladder carcinoma in situ. The authors investigated the correlation between tumor-associated macrophages infiltrating bladder carcinoma in situ and the response to intravesical bacillus Calmette-Guerin therapy. Materials and methods: The authors examined paraffin-embedded tissues from 41 patients with bladder carcinoma in situ who received intravesical bacillus Calmette-Guerin therapy. Tumor-associated macrophages were immunohistochemically stained by anti-CD68 monoclonal antibody. Results: The median number of tumor-associated macrophages infiltrating among cancer cells and the number in the lamina propria were 4 and 24, respectively. Recurrent carcinoma in situ was found in 4.8% of cases with a lower cancer cell tumor-associated macrophage count but in 47.6% of those with a higher cancer cell tumor-associated macrophage count (less than 4 vs. 4 or greater). Recurrence was found in 31.8% of patients with a lower lamina propria tumor-associated macrophage count but in 21.1% of those with a higher lamina propria tumor-associated macrophage count (less than 25 vs. 25 or greater). The median ratio of tumor-associated macrophages among cancer cells vs. in the lamina propria was 0.2. Recurrence-free survival was significantly better in patients with a lower cancer cell tumor-associated macrophage count (p = .0002). Those with a lower cancer cell-to-lamina propria tumor-associated macrophage ratio had a higher recurrence-free rate (p < .0001). Multivariate analysis revealed that the cancer cell tumor-associated macrophage count and the cancer cell-to-lamina propria tumor-associated macrophage ratio can be prognostic factors for bladder carcinoma in situ. Conclusions: The count of tumor-associated macrophages infiltrating the cancer area is useful for predicting the response of bladder carcinoma in situ to intravesical bacillus Calmette-Guerin instillation before treatment initiation. Although on univariate analysis TAMs are associated with other poor prognosticators, on multivariate analysis, TAMs appear only to be associated with MI and VI. TAMs may play a significant role in the biology of tumor progression of endometrial adenocarcinoma, but do not appear to be independent prognostic indicators of patient’s survival.

Prognostic Impact of Angiogenesis in Nonmuscle Invasive Bladder Cancer as Defined by Microvessel Density after Immunohistochemical Staining for CD34

Bladder cancer is the second most common malignancy of the urogenital region. The majority of bladder cancer deaths occur as a consequence of metastatic disease. Microvessel density (MVD), a surrogate marker for angiogenesis, has been shown to be predictive of progression and poor prognosis. The aim of this study was to evaluate the predictive value and prognostic significance of angiogenesis in human non muscle invasive bladder cancer (NMIBC) treated by BCG immunotherapy. The frozen sections of 28 non muscle invasive bladder cancer specimens were stained with CD34 antibody to label the vascular endothelium using the standard streptavidin-biotin immunoperoxidase method. Angiogenic activity was measured using microvessel count determined by the expression of vascular markers CD34.The prognostic significance of tumor stage, grade, loci number, tumor size, age and CD34 expression in determining the risk for recurrence was studied with both univariate and multivariate methods of analysis. According to univariate analysis of the prognostic significance for tumor stage, grade, tumor size, loci number, age and CD34 expression, in patients with NMIBC, the pT1 stage and high grade seem to be associated in a statistically significant manner with higher risk for recurrence (P=0.004, P=0.004, respectively). In the other hand, multivariate Cox regression’s analysis showed that microvessel density and multiplicity were independent predictor of recurrence after BCG immunotherapy (p=0.016, p=0.032, respectively). This study provides strong evidence that CD34 MVD is associated with recurrence after BCG immunotherapy. Independent studies, however, will be required on larger cohort to validate these findings.

Prognostic Value of Bcl-2 and Bax Tumor Cell Expression in Patients with Non Muscle-Invasive Bladder Cancer Receiving Bacillus Calmette-Guerin Immunotherapy

Apoptosis is the distinctive form of programmed cell death that complements cell proliferation in maintaining normal tissue homeostasis. The significance of constitutive apoptosis in the recurrence of Non Muscle Invasive Bladder Cancer has yet to be investigated. The aim of this study is to investigate the prognostic significance of Bax and Bcl-2 in terms of recurrence after BCG immunotherapy. Immunohistochemical analysis was performed on frozen biopsies to evaluate bcl-2 and Bax proteins expression in 28 cases of NMIBC. All patients with confirmed NMIBC were treated with intravesical BCG-immunotherapy. The follow up was performed for 26 months. The correlation between clinicopathological, immunohistochemical data and the response to BCG therapy was performed. Univariate analysis showed that, PT1 stage, High grade and Bax expression increased significantly the risk of recurrence (P = 0.015, P = 0.015 and P= 0.034 respectively). In addition, multivariate analysis selected the model involving stage, age, Bax and Bcl-2 expression as the best independent variables of recurrence. In conclusion, the expression of Bcl-2 and Bax in NMIBC could have a prognostic value in assessing the risk of recurrence after BCG immunotherapy. These findings require further investigations on larger cohort in order to ascertain new molecular markers of the response to BCG immunotherapy.

Tumor multiplicity is an independent prognostic factor of non-muscle-invasive bladder cancer treated with Bacillus Calmette-Guerin immunotherapy.

Background: Bacillus Calmette-Guerin (BCG) immunotherapy is regarded as the current treatment of choice for non muscle invasive bladder cancer (NMIBC), though its efficacy is limited by high recurrence and progression rate. Identification of factor prognosticators that might be helpful in discriminating between responders and nonresponders to BCG treatment is therefore of major clinical importance. The aim of this study is to evaluate the prognostic factors of recurrence after intravesical adjuvant BCG immunotherapy in patients with NMIBC. Methods: we retrospectively reviewed the clinical and pathologic data of primary NMIBC from 112 patients who were treated with transurethral resection followed by BCG-immunotherapy. Time follow-up was 30 months. The prognostic significance of tumor stage, grade, multiplicity, age, sex and smoking in determining the risk for recurrence after BCG therapy was studied with both univariate and multivariate methods of analysis. Results: According to univariate analysis of the prognostic significance for tumor stage, grade, loci number, sex, age and smoking, the pT1 stage and multiplicity seem to be associated in a statistically significant manner with higher risk for recurrence (P = 0.009, P = 0.011, respectively). In the other hand, multivariate analysis showed that only multiplicity was an independent significant prognosticator. Conclusion: Significant independent predictor for recurrence was multiplicity which offers important clinical information and may be a useful tool in the selection of suitable candidates for BCG-immunotherapy.

The Efficiency of the EORTC Scoring System for the Prediction of Recurrence and Progression of Non-muscle-invasive Bladder Cancer Treated by Bacillus Calmette-Guerin Immunotherapy

Abstract The authors determined the recurrence and progression at 1 year in patients with non-muscle-invasive urothelial carcinoma who underwent transurethral resection of bladder tumor (TURBT) and compared the results with the calculated risk according to the European Organization of Research and Treatment of Cancer (EORTC). Between 2002 and 2011, a total of 112 patients with NMIBC were treated with transurethral resection of bladder cancer. According to the EORTC scoring system, the patients were categorized in terms of number of tumors, tumor size, prior recurrence rate, T category, carcinoma in situ, and pathologic grade, and the scores were summed. According to the summed scores, the recurrence group and the progression group were divided into 3 subgroups: low, intermediate, and high risk, respectively. The recurrence rate and progression rate of each group were compared with the EORTC risk tables. The mean patient age was 63.9 years (range: 25–85) at diagnosis. Seventy-eight patients (68.4%) had a recurrent disease, 53 (47.3%) had a tumor larger than 3 cm in diameter, 55 (49.1%) had multiple lesions, 3 (0.26%) had carcinoma in situ, 44(39.3%) had stage T1 lesions, and 20 (17.8%) had a high-grade disease. The recurrence rates were 0, 14.2, 31.25, and 85.71% in groups with the predicted EORTC risks of 15, 24, 38, and 61%, respectively. There were 3 patients (0.2%) with progression of the diseases. The EORTC model successfully stratified recurrence and progression risks in this cohort. However, the discriminative ability of the EORTC tables decreased in these patients for progression.

Impact of Smoking intensity on Outcomes of Patients with Non Muscle Invasive Bladder Cancer Treated by BCG Immunotherapy

Abstract Background: Cigarette smoking is a well-known risk factor of bladder carcinogenesis. The clinical impact of smoking on bladder cancer recurrence and response to BCG immunotherapy remains unclear. We sought to investigate the effect of smoking intensity on bladder cancer response to BCG therapy, and the interactions between smoking and clinicopathological factors on bladder cancer recurrence. Methods: Clinical information was obtained from 81 smokers patients (smokers at diagnosis) with NMIBC treated with transurethral resection of the bladder tumor followed by BCG immunotherapy. The distribution of smoking intensity on patient age (≥60 years or <60 years), gender, tumor grade, tumor stage, carcinoma in situ, multiplicity and tumor size was assessed. The effect of cigarette smoking on cancer recurrence was estimated using Cox proportional hazard models and Kaplan–Meier analysis. Results: The results showed that smoking intensity was significantly associated with response to BCG immunotherapy (p = 0.010). Univariate Cox regression analysis of clinicopathologic characteristics showed that PT1 stage, tumor size more than 3 cm and smoking intensity significantly increased the risk of recurrence (respectively, p = 0.006; p = 0.008 and p = 0.012). These results were confirmed by Kaplan–Meier survival curves. In addition, multivariate analysis using Cox regression selected the model involving stage, tumor size and smoking intensity as the quasi-independent predictor of recurrence.Conclusion: These findings suggest that cigarette smoking is an independent predictor for patients with NMIBC. Although the current evidence supports a positive link between smoking intensity and the risk of recurrence on NMIBC treated by BCG immunotherapy, additional studies, are needed before definitive conclusions can be drawn.

Prognostic Significance of Heat Shock Protein 90 in Non Muscle Invasive Bladder Cancer Treated by BCG Immunotherapy

Background: The heat shock protein 90 (HSP90) is overexpressed and highly associated with poor prognosis in many malignancies. However, the role of HSP90 in Non muscle invasive bladder cancer (NMIBC) has not been thoroughly elucidated. The aim of this study is to investigate the relationship of HSP90 expression with the response to BCG immunotherapy of NMIBC. Methods: HSP90 expression was evaluated by immunohistochemistry in 43NMIBC samples. The relationships between HSP90 and the response to BCG immunotherapy were analyzed. The results were correlated with clinicopathological characteristics using univariate and multivariate analyses. Results: Univariate Cox regression analysis of baseline characteristics and HSP90 expression showed that no significant association was found with BCG immunotherapy response (p=0.894). In the other hand, multivariate Cox regression analysis showed that HSP90 protein is not an independent factor of tumor response after BCG immunotherapy (p=0.884). Conclusion: This study demonstrates that HSP90 could not be a useful prognostic marker for BCG treatment in NMIBC. We emphasize that this was a preliminary study and therefore further confirmation on a larger set of tissues is necessary.

Prognostic Value of Artificial Neural Network in Predicting Bladder Cancer Recurrence after BCG Immunotherapy

Background: Artificial neural network (ANN) has been used in medicine to predict either the treatment or the investigative outcomes. The aim of this study was to validate the use of ANN models for predicting recurrence in non muscle invasive bladder cancer (NMIBC) treated by Bacillus Calmette Guerin (BCG) immunotherapy. Materials and Methods: In this study, we developed a Multilayer Percepteron (MLP) based ANN to detect recurrence in NMIBC through the analysis of histopathologic data. The study includes 308 patients (mean age, 63.92 years; range, 31-92 years) who were treated with transurethral resection followed by BCG-immunotherapy. Time follow-up was 30 months. Results: In the test group, 39 out of 40 cases were correctly classified by the MLP base neural network with an optimum Mse error (0.02634). Only one case was classified as false positive, with no false negative results. The sensitivity, specificity, positive predictive and negative predictive values calculated from the output data were 96.66%, 100%, 100% and 90.9% respectively. Network can predict the outcome of 79% (34*100/35) of patients in the testing data set correctly. Conclusion: The proposed algorithm produced high sensitivity and specificity in predicting the recurrence in NMIBC after BCG immunotherapy compared to conventional statistical analysis. Therefore the use of ANNs will increasingly become the method of choice to calibrate complex medical models.

The Efficacy of Intravesical Bacillus Calmette-Guerin in the Treatment of Patients with pT1 Stage Non-muscle-invasive Bladder Cancer

Abstract Background: pT1 bladder urothelial carcinomas represent a heterogeneous group of tumors with different biologic behaviors, and identifying the subset of tumors that carries a high risk of disease recurrence and progression is therefore important. Induction and maintenance intravesical Bacillus Calmette-Guerin (BCG) has been proven to reduce tumour recurrence and progression. However, no markers are available to predict BCG response. The aim of this study is to evaluate the prognostic factors of stage in predicting recurrence after intravesical adjuvant BCG immunotherapy in patients with NMIBC. Methods: we retrospectively reviewed the clinical and pathologic data of primary NMIBC from 45 patients who were treated with transurethral resection followed by BCG-immunotherapy. Time follow-up was 30 months. The prognostic significance of clinicopathologics characteristics in determining the risk for recurrence after BCG therapy was studied with both univariate and multivariate methods of analysis. Results: univariate Cox regression analysis of clinicopathologic characteristics revealed that the rate of recurrence was statistically associated with tumor stage. Indeed, a significant concordance was noted between the EORTC s predicted risks and the actuarial recurrence rate of NMIBC at one year. On the other hand, multivariate analysis using Cox regression based on the AIC criteria and biological considerations, selected the score of recurrence as independent predictor of recurrence. Conclusion: The conventional clinicopathological factors used in EORTC model are relevant for the assessment of the outcome of pT1 stage bladder tumors treated by BCG immunotherapy. Management of pT1 bladder cancer patients remains one of the most difficult problems in urologic practice. At this time the decision to preserve the bladder or to perform a cystectomy depends on a number of clinicopathologic parameters, but none are able to sufficiently identify patients for the appropriate therapeutic modality. Additional studies using a more large scale of patients will be required to confirm our findings.

The Impact of Age and Clinical Factors in Non-muscle-invasive Bladder Cancer Treated with Bacillus Calmette Guerin Therapy

Abstract Background: Bladder cancer is a disease of older persons, the incidence of which is expected to increase as the population ages. Prognostic factors for local recurrence for patients with non-muscle invasive bladder cancer have not been fully established. The aim of our study was to determine the influence of age on the outcomes of non muscle invasive bladder (NMIBC) cancer treated with intravesical Bacillus Calmette–Guerin (BCG) therapy. Methods: We retrospectively reviewed the clinical and pathologic data of primary NMIBC from 112 patients who were treated with transurethral resection followed by BCG-immunotherapy. Time follow-up was 30 months. Clinocopathologic characteristics and response to BCG therapy were correlated with age using univariate and multivariate methods of analysis. Results: Univariate analysis showed that age analyzed as a categorical variable was not associated with other clinicopathological characteristics. On the other hand, multivariate analysis showed that only multiplicity, stage and tumor size were independent significant prognosticators. Conclusions: The results of our study have shown that aging has no impact on the outcomes of high-risk NMIBC treated by BCG immunotherapy.