Kazumasa Torimoto

Collagen type IV alpha 1 (COL4A1) and collagen type XIII alpha 1 (COL13A1) produced in cancer cells promote tumor budding at the invasion front in human urothelial carcinoma of the bladder

Current knowledge of the molecular mechanism driving tumor budding is limited. Here, we focused on elucidating the detailed mechanism underlying tumor budding in urothelial cancer of the bladder. Invasive urothelial cancer was pathologically classified into three groups as follows: nodular, trabecular, and infiltrative (tumor budding). Pathohistological analysis of the orthotopic tumor model revealed that human urothelial cancer cell lines MGH-U3, UM-UC-14, and UM-UC-3 displayed typical nodular, trabecular, and infiltrative patterns, respectively. Based on the results of comprehensive gene expression analysis using microarray (25 K Human Oligo chip), we identified two collagens, COL4A1 and COL13A1, which may contribute to the formation of the infiltrative pattern. Visualization of protein interaction networks revealed that proteins associated with connective tissue disorders, epithelial-mesenchymal transition, growth hormone, and estrogen were pivotal factors in tumor cells. To evaluate the invasion pattern of tumor cells in vitro, 3-D collective cell invasion assay using Matrigel was performed. Invadopodial formation was evaluated using Gelatin Invadopodia Assay. Knockdown of collagens with siRNA led to dramatic changes in invasion patterns and a decrease in invasion capability through decreased invadopodia. The in vivo orthotopic experimental model of bladder tumors showed that intravesical treatment with siRNA targeting COL4A1 and COL13A1 inhibited the formation of the infiltrative pattern. COL4A1 and COL13A1 production by cancer cells plays a pivotal role in tumor invasion through the induction of tumor budding. Blocking of these collagens may be an attractive therapeutic approach for treatment of human urothelial cancer of the bladder.

Two kinds of urinary continence reflexes during abrupt elevation of intravesical pressure in rats.

Urethral closure mechanisms during abrupt elevation of intravesical pressure (Pves) were investigated. Middle urethral contractile responses were observed during sneezing and passive Pves rise for 120 sec. Both responses were decreased when somatic nerves innervating the external urethral sphincter and the pelvic floor muscles were transected. In contrast, while transection of both pelvic and hypogastric nerves had no effect during sneezing, urethral responses during passive Pves rise were totally abolished or partially reduced after transection of pelvic or hypogastric nerves, respectively. Electrical stimulation of the abdominal muscles for 1 sec elevated Pves, and Pves inducing fluid leakage from the urethral orifice was lowered after somatic or pelvic nerves were cut, while hypogastric nerve transection showed smaller effects. These results indicate that at least two kinds of urinary continence reflexes close the middle urethra; one is preprogrammed to close irrespective of bladder afferent activity, the other is triggered by bladder afferent activity. During momentary stress events lasting within 1 sec, the striated muscle mainly contributes to urethral closure, while during events lasting a relatively long period, like passive Pves rise for 120 sec, both striated and smooth muscles are related to the prevention of urinary incontinence.

Integrative Assessment of Pretreatment Inflammation-, Nutrition-, and Muscle-Based Prognostic Markers in Patients with Muscle-Invasive Bladder Cancer Undergoing Radical Cystectomy

Objective: The present study evaluated the clinical relevance of an integrative preoperative assessment of inflammation-, nutrition-, and muscle-based markers for patients with muscle-invasive bladder cancer (MIBC) undergoing curative radical cystectomy (RC). Methods: The analysis enrolled 117 patients and the variables included age, body mass index (BMI), neutrophil-to-lymphocyte ratio, monocyte-to-lymphocyte ratio, platelet-to-lymphocyte ratio, modified Glasgow Prognostic Score (mGPS), prognostic nutritional index (PNI), Controlling Nutritional Status score, psoas muscle index (PMI), and peak expiratory flow (PEF). The correlations among the variables were evaluated and their prognostic values after RC were tested. Results: Three inflammation markers (ratios of blood cell counts) were positively correlated (p < 0.0001). The PNI and the BMI were positively correlated (p = 0.04), although they were inversely correlated with the three inflammation markers (p < 0.0001). Age was not significantly correlated with the inflammation markers and PMI, although older age was associated with lower PNI and lower PEF. The disease-specific survival was independently predicted by T4 tumor, positive N status, and decreased PNI. Overall survival was independently predicted by T4 tumor, mGPS, and pretreatment sarcopenia status. Conclusions: The inflammation-, nutrition-, and muscle-based markers would be useful risk assessment tools for MIBC.

Regulatory T Cells and Tumor-Associated Macrophages in the Tumor Microenvironment in Non-Muscle Invasive Bladder Cancer Treated with Intravesical Bacille Calmette-Guérin: A Long-Term Follow-Up Study of a Japanese Cohort

The clinical significance of regulatory T cells (Treg) and tumor-associated macrophages (TAM) in the tumor microenvironment of human bladder cancer remains unclear. The aim of this study is to explore their relevance to oncological features in non-muscle invasive bladder cancer (NMIBC). We carried out immunohistochemical analysis of forkhead box P3 (FOXP3, Treg maker), CD204 (TAM marker), and interleukin-6 (IL6) using surgical specimens obtained from 154 NMIBC patients. The Treg and TAM counts surrounding the cancer lesion and IL6-positive cancer cell counts were evaluated against clinicopathological variables. We focused on the ability of the Treg and TAM counts around the cancer lesion to predict outcomes after adjuvant intravesical Bacille Calmette–Guérin (BCG) treatment. High Treg counts were associated with female patients, older age, T1 category, and high tumor grade. TAM count was significantly correlated with Treg count and with IL6-positive cancer cell count. In our analysis of 71 patients treated with BCG, high counts of Treg and TAM were associated with shorter recurrence-free survival, and the former was an independent predictor of recurrence. Poor response to intravesical BCG was associated with Treg and TAM in the tumor microenvironment. Disrupting the immune network can be a supplementary therapeutic approach for NMIBC patients receiving intravesical BCG.

Lower Urinary Tract: Diabetes Mellitus-focused on Recent Experimental Results

Historically, it has been widely believed that urologic and neuropathic problems occur as late complications in the disease process of diabetes mellitus (DM). However, there is recent evidence that diabetic neuropathy can occur early in the disease process, with urologic manifestation. In the present article we focus on the complications of DM in the lower urinary tract (diabetic cystopathy and urethropathy).

Oncological outcome, complications, lower urinary tract symptoms, and health-related quality of life after low-dose-rate salvage brachytherapy for recurrent prostate cancer following primary radiotherapy: a report of 8 cases

Purpose We evaluated our experience with low-dose-rate salvage brachytherapy for local recurrence after primary prostate radiotherapy, and described the changes in lower urinary tract symptoms and health-related quality of life. Material and methods Between 2011 and 2016, eight men with local recurrence after primary prostate radiotherapy underwent iodine-125 salvage brachytherapy with a prescribed dose of 110 or 145 Gy. Recurrence-free survival was evaluated with a post-treatment prostate-specific antigen profile. The toxicity and changes in lower urinary tract symptoms and health-related quality of life during the follow-up were evaluated on the Common Terminology Criteria for Adverse Events version 4.0, International Prostate Symptom Score, Short Form-8, and Expanded Prostate Cancer Index Composite, respectively. Results The median follow-up was 12.2 months (range, 8.3-71.9) after salvage brachytherapy. Of all eight patients, two (25%) experienced treatment failure, one of whom developed left seminal vesicle recurrence 36 months after salvage brachytherapy for the right seminal vesicle recurrence, while the other developed bone metastases after 6 months. The International Prostate Symptom Scores peaked at 3 months, and returned to baseline by 6 months. The scores of all domains of health-related quality of life remained unchanged during the 12-month follow-up after salvage brachytherapy. Early grade ≤ 2 genitourinary toxicity was observed in five patients (63%), and late grade 2 gastrointestinal toxicity in one patient (13%) having persistent diarrhea. No patient required intermittent catheterization and no grade 3 or greater toxicity occurred during follow-up. Conclusions The present study is our experiment of eight patients undergoing salvage brachytherapy, suggesting that this modality is noninvasive, safe, and an effective salvage local treatment in selected patients. To our knowledge, this is the first study to evaluate lower urinary tract symptoms and health-related quality of life in the post-treatment period in prostate cancer patients.

URETHROPLASTY FOR COMPLICATED ANTERIOR URETHRAL STRICTURES

(Objectives) To compare efficacy and outcome of urethroplasty for complicated anterior urethral strictures. (Methods) Twelve patients, included 3 boys, with anterior urethral stricture underwent urethroplasty after the failure of either urethral dilatation or internal urethrotomy. We evaluated pre- and post-operative Q max and surgical outcome. (Results) Four patients were treated with end-to-end anastomosis, included a case of bulbar urethral elongation simultaneously, one patient was treated with augmented anastomotic urethroplasty, three patients were treated with onlay urethroplasty with prepucial flap, one patient was treated with tubed urethroplasty with prepucial flap (Ducket procedure) and three patients were treated with onlay urethroplasty with buccal mucosal graft. Postoperative Qmax improved in all patients without major complications and recurrence during follow-up periods ranging from 17 to 102 months (mean 55 months). (Conclusions) Urethroplasty is an effective therapeutic procedure for complicated anterior urethral stricture.

Relationship between tissue composition and biomechanics of the urinary bladder: effects of spinal cord injury on bladder tissue

Our previous studies have demonstrated that spinal cord injury and the associated urinary tract dysfunctions lead to significant changes in the bladder wall tissue mechanical behaviors. The present study made the first attempt to link biomechanical property changes of the bladder wall tissue and compositional changes at the molecular-level.

Effects of Diabetes Mellitus on the Biomechanical Properties of the Female Rat Urethra in the Passive State

Patients with diabetes mellitus (DM) suffer impaired lower urinary tract dysfunctions. The purpose of the current study was to evaluate the effects of DM on the passive biomechanical properties of the female rat urethra. DM was induced by injection of streptozotocin. Urethras were excised and mounted in an ex-vivo testing system. EDTA was added to the bath to inactivate smooth muscle. Continuous outer diameter measurements were made at proximal, middle, and distal portions of the urethra with a laser micrometer during stepwise increases of static, intraurethral pressure (0 to 20 mmHg). Compliance and beta stiffness were calculated from measured data. Healthy urethras served as controls. Statistical comparisons were made via ANOVA. The control tissue was most compliant proximally and decreased significantly along the length. This compliance gradient vanished with DM. A significant decrease in compliance and increase in beta stiffness was noted for 10 wk DM compared to controls. These findings suggest that DM has a large effect on the biomechanical properties of the urethra.Copyright

Effects of diabetes mellitus on the biomechanical properties of the female rat urethra

The effects of diabetes mellitus (DM) on bladder function are well described, but little is known of effects on the outlet function of the urethra. In the present study, biomechanical properties of urethras from DM and healthy female rats were compared. At 3, 5, and 10 weeks following streptozotocin induction of DM, urethras were excised from anesthetized rats maintaining in vivo length, mounted onto tees at in vivo length in an established ex vivo vascular testing system, and maintained in an oxygenated circulating bath of physiological media at 37/spl deg/C. Stepwise intraurethral pressure increments from 0-20 mmHg were applied and diameters were simultaneously recorded at proximal, mid, and distal positions. Urethras from age-matched rats served as controls. There was a proximal-distal decreasing gradient in compliance in normal urethras, and a progressive increase in beta stiffness from 3-10 weeks following DM in the mid and proximal regions to values indistinguishable from the distal. Increasing urethral stiffness in DM results in a collapse of the compliance gradient leading to increased outlet resistance for a bladder in which contractility is already compromised. Thus, DM effects on the urethra interact with those on the bladder to further compromise lower urinary tract function.