Tumor Infiltrating Neutrophils Are Enriched in Basal-Type Urothelial Bladder Cancer.

BACKGROUND: Urothelial bladder cancers (UBCs) are distinct in two main molecular subtypes, namely basal and luminal type. Subtypes are also diverse in term of immune contexture, providing a rationale for patient selection to immunotherapy. METHODS: By digital microscopy analysis of a muscle-invasive BC (MIBC) cohort, we explored the density and clinical significance of CD66b+ tumor-associated-neutrophils (TAN) and CD3+ T cells. Bioinformatics analysis of UBC datasets and gene expression analysis of UBC cell lines were additionally performed. RESULTS: Basal type BC contained a significantly higher density of CD66b+ TAN compared to the luminal type. This finding was validated on TCGA, GSE32894 and GSE124305 datasets by computing a neutrophil signature. Of note, basal-type MIBC display a significantly higher level of chemokines (CKs) attracting neutrophils. Moreover, pro-inflammatory stimuli significantly up-regulate CXCL1, CXCL2 and CXCL8 in 5637 and RT4 UBC cell lines and induce neutrophil chemotaxis. In term of survival, a high density of T celsl and TAN was significantly associated to a better outcome, with TAN density showing a more limited statistical power and following a non-linear predicting model. CONCLUSIONS: TAN are recruited in basal type MIBC by pro-inflammatory CKs. This finding establishes a groundwork for a better understanding of the UBC immunity and its relevance.

How to improve patient selection for neoadjuvant chemotherapy in bladder cancer patients candidate for radical cystectomy and pelvic lymph node dissection.

PURPOSE: To improve patient selection for neoadjuvant chemotherapy (NAC) before radical cystectomy (RC) in bladder cancer patients (BCa). METHODS: Retrospective evaluation of 1057 patients with cT2-4N0M0 BCa treated with RC and pelvic lymph node dissection between 1990 and 2018 at 3 referral centers. Adverse pathologic features (APF) were defined as pT3-pT4/pN + disease at RC. A regression tree model (CART) was used to assess preoperative risk group classes. A multivariable logistic regression (MVA) was performed to identify predictors of APF at RC. RESULTS: Median age was 70 years and most of the patients were men (83%). Of the 1057 patients included in our study, 688 (65%) had APF. CART analysis was able to stratify patients into 3 risk groups: low (cT2 and single disease, odds ratio [OR] 0.62), intermediate (cT2 and multiple disease, OR 1.08), and high (cT3-cT4, OR 1.28). On MVA APF were associated with variant histology (odds ratio [OR] 3.97, 95% confidence interval [CI] 1.46-10.83, p = 0.007), multifocality at TUR (OR 2.56, CI 1.27-5.17, p = 0.09), completeness of resection (OR 0.47, CI 0.23-0.96, p = 0.04) and clinical extravesical disease (OR 3.42, CI 1.63-7.14, p = 0.001). CONCLUSION: We defined three pre-operative risk classes. Our results indicate that patients with a cT3-T4 disease are those who might benefit more from NAC whereas those with T2 single disease should be those to whom NAC probably shouldn't be proposed. Given the high rate of understaging in BCa patients, NAC can be proposed in selected cases of cT2/multifocal disease.

Hyper-Activation of STAT3 Sustains Progression of Non-Papillary Basal-Type Bladder Cancer via FOSL1 Regulome.

Urothelial bladder cancer (UBC) are classified into luminal and basal subtypes showing distinct molecular features and clinical behaviour. Recent in silico data have proposed the activation on the Signal Transducer and Activator of Transcription 3 (STAT3) as relevant transcription factor in UBC. To answer this question, we have combined the retrospective analysis of clinical samples, functional assays on cell lines, interrogation of public UBC datasets and a murine model of basal-type UBC. Immunohistochemistry on a retrospective UBC cohort uncovered that STAT3 Y705 phosphorylation (pSTAT3) is significantly increased in infiltrating basal-type UBC compared to luminal UBC. In vitro, STAT3 silencing in UBC cell lines significantly reduced tumor cell viability and invasion. Gene expression profile of UBC cell lines combined with the analysis of the Cancer Genome Atlas (TCGA) and GSE32894 UBC datasets showed that increased expression of a set of STAT3 targets predicts basal-type, propensity to local progression and worse prognosis. MYC and FOSL1 represent relevant STAT3 downstream targets, as validated by their co-localization in pSTAT3+ UBC cancer cells. These findings were largely reproduced in the BBN-induced murine model of basal-type UBC. Of note, FOSL1 protein resulted strongly expressed in the non-papillary UBC pathway and FOSL1-regulated transcripts were significantly enriched in the transition from NMIBC to MIBC, as indicated by the interrogation of the GSE32894 dataset. The blockade of the STAT3 pathway might represent a novel treatment option for these neoplasms. Monitoring pSTAT3 and the downstream targets, particularly FOSL1, could provide meaningful levels of UBC stratification.

The impact of completeness of last transurethral resection of bladder tumors on the outcomes of radical cystectomy.

PURPOSE: To evaluate the role of a complete transurethral resection of bladder tumors (c-TURBT) on oncological outcomes after radical cystectomy (RC) and its relationship with adverse pathological features. METHODS: We retrospectively analyzed data of 727 patients treated with RC and bilateral pelvic lymph node dissection at three tertiary referral centers. Possible c-TURBT was reported by the treating surgeon. Multivariable Cox regression analyses were used to assess the relationship of c-TURBT and survival outcomes after surgery in 1:1 propensity score-matched cohort adjusted for age and gender. Moreover, multivariable logistic regression (MVA) was built to predict the relationship between c-TURBT and pT3-T4 stages at RC, lymph node invasion (LNI) and positive soft tissue surgical margin (STSM). RESULTS: A total of 433 (60%) patients received a c-TURBT. 3.0% of patients with a c-TURBT achieved a pT0-pTa-pTis status vs. 2.0% of patients with incomplete TURBT. At multivariable Cox regression analyses, c-TURBT was not associated with survival outcomes. At MVA, incompleteness of TURBT was significantly associated with a pT3-T4 stage [odds ratio (OR) 8.04, 95% confidence interval (CI) 2.33-27.67, p = 0.001]. No significant association was found between c-TURBT, LNI and STSM. CONCLUSION: We found a low rate of achievement of pT0 stage at RC. An incomplete TURBT before RC represented a predictor of pT3-T4 stages, but no effect of a c-TURBT was shown on survival outcomes. Given the current inadequacy of clinical staging strategies with more than 50% of extravesical disease being under-staged, our results could improve patients selection for NAC, driving the decision-making in doubtful cases.

Prognostic role of delay before radical cystectomy: retrospective analysis of a single-centre cohort with 376 patients.

BACKGROUND: According to International Guidelines radical cystectomy (RC) should not be delayed over 90 days to prevent the risk of intercurrent progression and worse survival. Nevertheless, such a recommendation relies on a few retrospective studies reaching non-univocal conclusions. Aim of the present study was to investigate if the latency between diagnosis and cystectomy (LDC) is related to prognosis after RC. METHODS: Retrospective analysis of database collecting complete information on patients undergone RC at single institution since 2004. The cases with an LDC <15 or >360 days or submitted to neo-adjuvant chemotherapy or with distant metastasis were excluded. Uni- and multivariate analyses assessed the relationship between LDC upstaging, progression-free and overall survival. RESULTS: The data of 376 patients were analyzed; mean/median LDC was 83/76 days and 124 patients (33%) had LDC>90 days. LDC was shorter in younger patients with first diagnosis of more advanced BC at clinical staging; accordingly, patients with LDC<90 days had more advanced disease also at final pathology. Prevalence of pathological upstaging was 37%; in case of upstaging LDC was 75 days vs. 72 days (P=0.4629). Multivariable regression models adjusted for pathological local and lymph nodal stage showed that LDC, continuous or dichotomized at 30/60/90/120/180/240 days was not related to progression-free or overall survival. The retrospective design of study is the main limitation of the study. CONCLUSIONS: In our experience nor the risk of upstaging, neither survival after RC were related to LDC. Even if these results should not discourage any effort to perform surgery expeditiously, the window of opportunity for RC might not be delimited by a predetermined threshold.

Features of Ipsilateral Renal Recurrences After Partial Nephrectomy: A Proposal of a Pathogenetic Classification.

BACKGROUND: Poor data are reported on the pathogenesis of ipsilateral relapse (IR) after partial nephrectomy (PN). The objective of this study was to investigate features of IR after PN with the intention to propose a pathogenetic classification. MATERIALS AND METHODS: Retrospective consultation of an institutional database that stores the data of 683 patients submitted to PN since 1993. The clinical, radiological, and follow-up data of the cases submitted to salvage nephrectomy due to an IR were analyzed. The slides of the sections from the tumor-parenchyma interface of PN and the bed of resection from the specimen of nephrectomy were reviewed. RESULTS: Eighteen patients were submitted to salvage nephrectomy for an IR. In 12 cases the IR harbored into the site of PN and a mixture of cancer cells and granulomatous reaction was found at the resection bed (IR type A). In the remaining 6, in microscopy of the resection bed was found only fibrosis: 3 of these cases had a clear-cell renal cell carcinoma (RCC) with diffuse microvascular embolization and the relapse in the same portion of the kidney of the primary tumor (IR type B); the other 3 had a non-clear-cell RCC and the primary and relapsing tumors were located in distinct portions of the kidney (IR type C). Six patients (4 IR type A, 2 type B) had a further progression and 5 of them died due to RCC. CONCLUSION: More frequently an IR is due to the incomplete resection of the primary tumor (IR type A), in a minority of the cases to the local spread of the tumor by microvascular embolization (IR type B), or true multifocality (IR type C). The prognosis of IR not due to multifocality (type A and B) is poor, despite salvage nephrectomy.

Features, risk factors and clinical outcome of "very late" recurrences after surgery for localized renal carcinoma: A retrospective evaluation of a cohort with a minimum of 10 years of follow up.

OBJECTIVE: To evaluate the features and the predictors of "very late" recurrences after surgery for localized renal cell carcinoma. METHODS: Since 1983, an institutional database with data of more than 2300 consecutive patients treated for renal cancer has been prospectively maintained. Patients N0 /Nx M0 followed for a minimum of 10 years without recurrences were retrieved. The site, time and treatment of recurrences observed afterwards were recorded, and the predictors were investigated by Cox regression analysis. RESULTS: A total of 554 patients (231 women, 323 men; age 59.3 ± 11.6 years) followed for a mean/median time of 15.1/13.6 years (range 10.0-34.1 years) were analyzed. A recurrence was observed in 26 patients (4.6%) after a mean/median interval of 13.3/12.3 years (range 10.5-30.2 years). The pathological stage 2/3 was the only independent predictor of recurrence (P = 0.003), and it was related also to the latency of recurrence (mean/median latency 15.4/14.0, 11.4/10.8 and 12.5/12.0 years, respectively, for stage 1, 2 and 3; P < 0.005 for stage 1 vs stage 2 or 3). The contralateral kidney was the most frequent site of relapse in patients with stage pT1, whereas multiple sites were more frequent for stage pT2 and pT3. CONCLUSIONS: The risk of a "very late" recurrence of renal cancer is approximately 5%, and it depends on the pathological stage. For stage pT1, the kidney/s should be surveilled for indefinite time, preferably by ultrasound to reduce the X-ray exposition; for stage pT2 and pT3, the abdomen and the lungs should be monitored, by computed tomography scan during the first years, and then by abdominal ultrasound and chest X-ray.

Comparison of Perioperative Morbidity of Radical Cystectomy With Neobladder Versus Ileal Conduit: A Matched Pair Analysis of 170 Patients.

BACKGROUND: The objective of this study was to evaluate if use of an ileal conduit (IC) versus a neobladder (NB) during radical cystectomy (RC) can play a role in the morbidity of the surgical procedure. PATIENTS AND METHODS: Since 2001 our institution has prospectively maintained a database of more than 500 patients who have undergone RC. The records of 258 patients who received an RC and IC and 121 who received an RC and NB were reviewed for the present study. Using a binary logistic regression model a propensity score was generated for the following factors: sex, age, smoking habit, Charlson score, American Society of Anesthesiology score, preoperative hydronephrosis, and pathologic T stage. Two groups of patients with similar propensity scores were created with a ratio of 1:1, one group who received an IC and another who received an NB. The following features were compared between the 2 groups: blood loss, intraoperative complication rate, time to oral intake resumption, postoperative complication rate (overall, Clavien classification 1-2, Clavien classification 3-5, related to RC, related to urinary reconstruction). RESULTS: In total, 170 patients were selected; 85 with NB, 85 with IC, and the 2 groups were well matched according to the desired features. No differences were noted for all of the compared features. CONCLUSION: The morbidity of RC does not seem to be affected by the choice of IC versus NB.

[Differences in morbidity of neobladder versus ileal conduit: a matched pair analysis on 134 patients]. / Differenza di morbidità fra neovescica ortotopica econdotto ileale. Matched pair analysis su 134 pazienti.

OBJECTIVE: The objective of this study is to evaluate whether performing an ileal conduit (IC) versus a neobladder(NB) during radical cystectomy (RC) can play a role on the morbidity of the surgical procedure. MATERIALS AND METHODS: At our institution since 2001, a database collecting the data of more than 450 patients who have undergone RC is perspectively maintained.The records of 246 patients submitted to RC and IC and 120 to RC and NB have been reviewed for the present study. By a binary logistic regression model, a propensity score was generated joining these factors: gender,age, smoking habit, Charlson' comorbidity score, platelet antiaggregants or oral anticoagulant therapy, ASA(American Society of Anesthesiologists) score, hydronephrosis, pathologic T stage, and lymphadenectomy. A group of patients submitted to IC and another to NB having the closer propensity score were created with a matching ratio of 1:1. The following features were compared between the two groups: operative time, blood loss,intraoperative complication rate, and postoperative complications rate (overall, clavien 1-2, clavien ≥, related toRC, relate to urinary reconstruction).Results: One hundred thirty-four patients were selected, 67 submitted to NB, 67 to IC, well matched according to the desired features. No differences were noted for all the compared features.Conclusion: The morbidity of RC does not seem to be dependent on the choice to perform IC versus NB.

[Differences in morbidity of neobladder versus ileal conduit: a matched pair analysis on 134 patients]. / Differenza di morbidità fra neovescica ortotopica econdotto ileale. Matched pair analysis su 134 pazienti.

OBJECTIVE: The objective of this study is to evaluate whether performing an ileal conduit (IC) versus a neobladder(NB) during radical cystectomy (RC) can play a role on the morbidity of the surgical procedure. MATERIALS AND METHODS: At our institution since 2001, a database collecting the data of more than 450 patients who have undergone RC is perspectively maintained.The records of 246 patients submitted to RC and IC and 120 to RC and NB have been reviewed for the present study. By a binary logistic regression model, a propensity score was generated joining these factors: gender,age, smoking habit, Charlson' comorbidity score, platelet antiaggregants or oral anticoagulant therapy, ASA(American Society of Anesthesiologists) score, hydronephrosis, pathologic T stage, and lymphadenectomy. A group of patients submitted to IC and another to NB having the closer propensity score were created with a matching ratio of 1:1. The following features were compared between the two groups: operative time, blood loss,intraoperative complication rate, and postoperative complications rate (overall, clavien 1-2, clavien ≥, related toRC, relate to urinary reconstruction).Results: One hundred thirty-four patients were selected, 67 submitted to NB, 67 to IC, well matched according to the desired features. No differences were noted for all the compared features.Conclusion: The morbidity of RC does not seem to be dependent on the choice to perform IC versus NB.