Intratumoral CCR5+ neutrophils identify immunogenic subtype muscle-invasive bladder cancer with favorable prognosis and therapeutic responses.

Our previous studies revealed tumor-infiltrating neutrophils (TINs) played dichotomous roles in different cancers, indicating diverse TINs subtypes might orchestrate anti-tumor immunity or immune evasion, respectively. This study aimed to investigate the clinical significance and immune characteristics of CCR5+TINs in muscle-invasive bladder cancer (MIBC). Two hundred and fifty-seven MIBC patients from two clinical centers and 95 fresh MIBC samples were included. CCR5+TINs were stained by immunohistochemistry, and the relationship between patients' clinic-pathological features and prognosis was evaluated, respectively. Immunohistochemistry and flow cytometry were applied to assess the immune features of CCR5+TINs and their correlations with other immune cells. In vitro study was conducted to estimate immune characteristics of CCR5+TINs and their predictive potential for pembrolizumab therapeutic response. In the two MIBC cohorts, we found that high CCR5+TINs infiltration could predict better overall survival (OS, P= .032, 0.039) and recurrence-free survival (RFS, P= .001, 0.006) and be associated with survival benefit from adjuvant chemotherapy (ACT, P< .001 for OS and P= .022 for RFS, respectively) in merely pT2N0 MIBC. Maraviroc could partly reduce IFN-γ secretion by CCR5+TINs (P< .001). CCR5+TINs correlated with higher expression of effector molecules within CD8+T cells. Notably, pembrolizumab treatment could only elevate the apoptosis status of tumor cells in the CCR5+TINs high subgroup (P < .001), other than CCR5+TINs low subgroup (P= .481). Our results indicate that CCR5+TINs could prime anti-tumor immune response through autonomous IFN-γ release, thus leading to favorable prognosis and superior therapeutic response to ACT and immunotherapy in MIBC.

Responses of bladder smooth muscle to the stretch go through extracellular signal-regulated kinase (ERK)/p90 ribosomal S6 protein kinase (p90RSK)/Nuclear factor-κB (NF-κB) Pathway.

AIMS: The present study was designed to study changes and its potential mechanisms in human bladder smooth muscle subjected to stretch. METHODS: Bioinformatics analyses including differential expression analysis, overrepresentation enrichment analysis, principal component analysis, and weighted gene coexpression network analysis were used to analyze a microarray dataset (GSE47080) of partial bladder outlet obstruction (pBOO) in rat to find the potential changes of gene expressions. Bladder from pBOO model and human bladder smooth muscle cells (HBSMCs) subjected to sustained prolonged stretch were collected for Western blot analysis, real-time polymerase chain reaction, and fluorescence analysis to verify the changes of gene expressions and preliminarily study the potential role of signaling pathway regulation in treatment of pBOO. RESULTS: The bioinformatics analysis showed that chronic obstruction activated mitogen-activated protein kinase pathway and changed cytoskeleton structure in bladder smooth muscle. In in vivo experiments in mice, pBOO was verified by cystometry. Partial BOO activated the extracellular signal-regulated kinase (ERK)/p90 ribosomal S6 protein kinase (p90RSK)/nuclear factor-κB (NF-κB) signaling pathway in DM. The messenger RNA (mRNA) expressions of contractile phenotypic proteins increased after pBOO. In in vitro experiments of HBSMCs, mechanical stretch activated ERK/p90RSK/NF-κB in HBSMCs in a time-dependent manner. The mRNA expressions of α-smooth muscle actin and SM22 also increased and filamentous actin (F-actin) polymerization was enhanced as well. Inhibition of ERK/p90RSK/NF-κB pathway reversed mechanical stretch-induced changes of contractile phenotypic expression and F-action polymerization. CONCLUSIONS: Continuous stretch increases expressions of contractile phenotypic proteins and promotes the polymerization of F-actin. This process partially goes through ERK/p90RSK/NF-κB pathway.

Tumor infiltrating mast cells determine oncogenic HIF-2α-conferred immune evasion in clear cell renal cell carcinoma.

PURPOSE: Hypoxia-inducible factor 2α (HIF-2α) overexpression leads to activation of angiogenic pathways. However, little is known about the association between HIF-2α expression and anti-tumor immunity in clear cell renal cell carcinoma (ccRCC). We aimed to explore how HIF-2α influenced the microenvironment and the underlying mechanisms. EXPERIMENTAL DESIGN: We immunohistochemically evaluated immune cells infiltrations and prognostic value of HIF-2α expression in a retrospective Zhongshan Hospital cohort of 280 ccRCC patients. Fresh tumor samples, non-tumor tissues and autologous peripheral blood for RT-PCR, ELISA and flow cytometry analyses were collected from patients who underwent nephrectomy in Zhongshan Hospital from September 2017 to April 2018. The TCGA KIRC cohort and SATO cohort were assessed to support our findings. RESULTS: We demonstrated that ccRCC patients with HIF-2αhigh tumors exhibited reduced overall survival (p = 0.025) and recurrence-free survival (p < 0.001). Functions of CD8+ T cells were impaired in HIF-2αhigh patients. In ccRCC patients, HIF-2α induced the expression of stem cell factor (SCF), which served as chemoattractant for mast cells. Tumor infiltrating mast cells impaired anti-tumor immunity partly by secreting IL-10 and TGF-ß. HIF-2α mRNA level adversely associated with immunostimulatory genes expression in KIRC and SATO cohorts. CONCLUSIONS: HIF-2α contributed to evasion of anti-tumor immunity via SCF secretion and subsequent recruitment of mast cells in ccRCC patients.

Development and prospective validation of a novel weighted quantitative scoring system aimed at predicting the pathological features of cystic renal masses.

OBJECTIVES: To develop and prospectively validate a novel weighted quantitative scoring system based on CT findings, namely, the renal cyst index (RCI), aimed at preoperatively predicting the pathological features of cystic renal masses (CRMs). METHODS: The RCI was based on four critical features of CRMs: the cyst wall, septal, nodule, and cyst contents. These parameters were scored with 1, 2, or 3 points. Weight coefficients for these parameters were determined by the multivariable logistic regression. The odds ratio (OR) and 95% confidence interval (95% CI) were used to summarise the results. The RCI was defined as the sum of these four weight coefficients. Malignancy risk prediction models were built based on the retrospective evaluation of 441 patients. We also compared the prediction ability of the RCI with the Bosniak classification in the 441 patients and applied these novel models to 152 masses resected in our institution to prospectively validate the efficiency of the RCI. RESULTS: The wall point (OR = 5.71 [95% CI = 1.734-18.808, p = 0.004, point = 2], OR = 12.665 [95% CI = 3.750-42.770, p < 0.001, point = 3]), septal point (OR = 3.325 [95% CI = 1.272-8.692, p = 0.014, point = 3]), nodule point (OR = 4.588 [95% CI = 1.429-14.729, p < 0.001, point = 2], OR = 17.032 [95% CI = 5.017-57.820, p = 0.010, point = 3]), content point (OR = 22.822 [95% CI = 1.041-495.995, p = 0.047, point = 2], OR = 2.723 [95% CI = 1.296-10.696, p = 0.015, point = 3]), and RCI (OR = 1.247 [95% CI = 1.197-1.299, p < 0.001]) were significantly associated with malignancy. Masses with an RCI < 6 were regarded as benign masses; masses with an RCI ≥ 10 were regarded as malignant masses. The malignancy risk of masses with an RCI > 6 but < 10 were determined by a nomogram. The prediction ability of the RCI was significantly superior to the Bosniak classification for Bosniak IIF and III masses (AUC: 0.912 vs. 0.753, p = 0.001). The RCI also accurately predicted the pathological features of 152 masses. CONCLUSION: The RCI is a reliable quantitative scoring system in predicting the malignancy risk of CRMs, and it outperformed the Bosniak classification system in some ways. KEY POINTS: • The renal cyst index (RCI) is a useful weighted quantitative classification system based on CT findings for diagnosing cystic renal masses. • The RCI outperforms the Bosniak classification system in some ways, especially for Bosniak IIF and III masses. • Masses with an RCI < 6 can be regarded as a simple cyst, while those with an RCI > 10 can be regarded as malignant masses.

Short interval of biopsy to robotic-assisted laparoscopic radical prostatectomy does not render any adverse effects on the perioperative outcomes.

The aim of our study was to investigate the effects of prostate biopsy on perioperative outcomes of robotic-assisted laparoscopic prostatectomy (RALP).A total of 181 patients who underwent the RALP in our institution have been retrospectively reviewed, patients were divided into different groups according to the interval of biopsy to RALP and core numbers of biopsy. Perioperative outcomes including estimated blood loss (EBL), operative time (OT), surgical margin status, postoperative drainage, hospital stay, and perioperative complications were served as endpoints.Interval of biopsy to RALP was not significantly correlated with any perioperative outcomes, while the biopsy core numbers had significant correlation with the EBL. In logistic regression analysis, the biopsy core numbers were associated with higher risk of positive surgical margins. Body mass index (BMI) was also a significant factor related to OT.Delay of the RALP after biopsy was not applicable in the era of RALP and surgeons could be more freely in selecting the time of RALP. Besides, further studies should focus on how to improve the diagnostic efficiency of prostate cancer without increasing the incidence of surgical complications.

An optimized transurethral catheterization cystometry in mice and comparison with classic suprapubic catheterization cystometry.

AIMS: The aim of this study is to establish an optimized, minimally invasive transurethral catheterization cystometry (TUCC) and a novel urethral pressure profile (UPP) measurement for mice. METHODS: The optimized TUCC and the UPP measurement were first established. This optimized TUCC was then performed in 16 anesthetized female mice and compared with the suprapubic catheterization cystometry (SCC) in parallel after suprapubic catheters implantation (SCI; on zero, third, and seventh day, respectively). Finally, the optimized TUCC and novel UPP measurement were applied to investigate in another eight mice of partial bladder outlet obstruction (pBOO) model. The urodynamic parameters including micturition pressure (MP), basal pressure (BP), threshold pressure (TP), bladder capacity (BC), micturition volume (MV), residual urine (RV), bladder compliance (COM), maximum urethral pressure (MUP), bladder pressure curve and UPP were recorded. Statistical cross-comparisons of parameters for two kinds of cystometries and pBOO model were performed. RESULTS: Compared with the optimized TUCC before SCI, the MV, RV, BC, and MP decreased significantly on the seventh day after SCI (270.4-132.5 µL, 46.13-20.09 µL, 316.4-152.5 µL, 30.01-24.34 cmH2 0, respectively). After SCI, the BP, MP, TP, MV, RV, BC, and COM showed no significant difference between the TUCC and SCC at the same time point. The MUP increased significantly after pBOO operation (19.1-46.6 cmH2 0, P < 0.05). CONCLUSIONS: The minimally invasive TUCC along with UPP measurement could be widely applied to study the bladder function of mice as a feasible, repeatable, and accessible method.

The zero ischemia index (ZII): a novel criterion for predicting complexity and outcomes of off-clamp partial nephrectomy.

PURPOSES: Although several anatomical classification systems that aimed to standardize the description of renal tumors were previously reported, a special classification system is required to help predict the complexity and perioperative outcomes of off-clamp nephron-sparing surgery (NSS). We developed a novel criterion-zero ischemia index (ZII), aiming to help predict the perioperative outcomes after off-clamp NSS and guide patient selection. METHODS: We retrospectively evaluated 149 patients between June 2009 and July 2014 in our institution who underwent off-clamp NSS with available computed tomography images. ZII was defined as the product of the tumor diameter and depth within renal parenchyma. ZII was then analyzed to investigate its association with perioperative outcomes. A specific ZII score was further selected to best guide patient selection in off-clamp NSS. RESULTS: ZII was significantly associated with estimated blood loss >500 mL (OR 1.270, 95% CI 1.036-1.557, p = 0.021), operative time >2 h (OR 1.286, 95% CI 1.051-1.573, p = 0.014), surgical complications (OR 1.251, 95% CI 1.035-1.511, p = 0.020), overall complications (OR 1.208, 95% CI 1.016-1.436, p = 0.032), and >10% decrease in estimated glomerular filtration rate (OR 1.362, 95% CI 1.045-1.776, p = 0.022). Patients with ZII > 6 may bear a higher risk of hemorrhage, perioperative complications, and a longer operative time, if they underwent an off-clamp NSS, compared to those with ZII ≤ 6. CONCLUSIONS: The ZII is a novel and readily measurable criterion which can help predict renal complexity of tumor and risk of perioperative outcomes after off-clamp NSS. ZII = 6 is established as a preliminary threshold for patient selection of off-clamp NSS. A more robust criterion is to be validated with more samples in a prospective study.

Number of Renal Columns Invaded by Tumor: A Novel Parameter for Predicting Complexity and Outcomes of Off-Clamp Open Partial Nephrectomy.

BACKGROUND: The number of renal columns invaded by tumor (NRC) can determine the tumor complexity and perioperative outcomes of off-clamp open partial nephrectomy (OPN). We aimed to propose a novel and simple scoring system and examine the relationship between NRC and perioperative outcomes after off-clamp OPN. STUDY DESIGN: We retrospectively evaluated 202 patients with contrast-enhanced CT imaging who underwent off-clamp OPN between January 2008 and November 2014. The number of renal columns invaded by tumor was correlated to baseline demographics and perioperative outcomes as a categorical variable using multivariable logistic regression analysis. RESULTS: The mean tumor size was 2.5 cm (±1.3 cm). The number of renal columns invaded by tumor had the highest correlation coefficients with estimated blood loss (coefficient = 0.485; p < 0.001), operative time (coefficient = 0.310; p < 0.001), postoperative drainage (coefficient = 0.307; p < 0.001), and hospital length of stay (coefficient = 0.144; p = 0.041). Multivariable logistic regression demonstrated NRC to be an independent predictor of estimated blood loss ≥500 mL, postoperative renal function, operative time, and surgical complications. This predictive ability of NRC was superior to the R.E.N.A.L. (radius exophyic/endophytic nearness anterior/posterior location) score and PADUA (preoperative aspects and dimensions used for an anatomical) score. Spearman correlation coefficient of NRC calculations between the 2 observers was 0.941 (p < 0.001). CONCLUSIONS: The number of renal columns invaded by tumor is a novel, intuitive, and practical parameter that could be used to quantify renal tumor complexity and predict the risk of perioperative outcomes after off-clamp OPN. Specifically, NRC correlates with estimated blood loss, operative time, surgical complications, renal function, and conversion to on-clamp. In the future, NRC can be integrated with other parameters, such as tumor location, to assemble a new scoring system.

Mini-flank supra-12th rib incision for open partial nephrectomy for renal tumor with RENAL nephrometry score ≥10: an innovation of traditional open surgery.

The skill of supra-12th rib mini-flank approach for open partial nephrectomy (MI-OPN) provides an advanced operative method for renal tumor. Compared with laparoscopic and robotic surgery, it may be a feasible selection for the complex renal tumors. We describe our techniques and results of MI-OPN in complex renal tumors with high RENAL nephrometry score (RENAL nephrometry score ≥10). Fifty-five patients diagnosed with renal tumors between January 2009 and July 2013 were included in this study. Eligibility criteria comprised of patients with complex renal tumor (RENAL score ≥10) being candidates for partial nephrectomy (PN). All patients received MI-OPN and all surgeries were performed by a single urologist. The preoperative workup comprised of medical history, physical examination, and routine laboratory tests. Serum creatinine was recorded preoperatively and 2 to 3 months after operation. Operative time, ischemia time, blood loss, operative and postoperative complications, renal function, and pathology parameters were recorded. MI-OPN was successfully performed in all cases. Mean tumor size was 4.7 cm (range: 2.5-8.1). Mean warm ischemia time was 28.1 minutes (range: 21-39), mean operative time was 105 minutes (range: 70-150) and mean estimated blood loss was 68 mL (range: 10-400). Mean postoperative hospital stay was 6.5 days (range: 5-12). Postoperative complications were found in 3 patients (5.5%). The mean pre- and postoperative serum creatinine levels were 76.2 µmol/L (range: 47-132) and 87.1 µmol/L (range: 61-189) with significant difference (P = 0.004). The mean pre- and postoperative estimated glomerular filtration rate (eGFR) were 91.5 (range: 34-133) and 82.5 (range: 22-126.5), respectively with significant difference (P = 0.024). In an average follow-up of 19.9 months (range: 8-50), no local recurrence or systemic progression occurred. In conclusion, MI-OPN can combine the benefits of both minimal invasive and traditional open partial nephrectomy (OPN) techniques with a smaller incision. It is an innovation of traditional OPN and suitable for the complex renal tumors with high RENAL nephrometry score safely and effectively.