Normal Bladder Mucosa Research Articles

A four-gene model for prognostic prediction in bladder urothelial carcinoma

BackgroundBladder urothelial carcinoma (BLCA) is one of the most common malignant tumors in urinary system worldwide. High possibility of recurrence and progression leads to poor prognosis, revealing the significant role of long-term postoperative monitoring to the patients. However, effective noninvasive diagnosis is currently limited. Materials and methodsDifferentially expressed genes (DEGs) were analyzed using R-packages. Functional enrichment analyses were performed on Metascape. The prognostic model was established by multi-step cox regression and evaluated by survival plots and receiver operating characteristic (ROC) curves. The nomogram was then constructed based on three identified prognostic factors. C-index and calibration curves were calculated to testify the capacity for predicting survival possibility of BLCA patients. The transcription levels of model genes were further verified in a gemcitabine-resistant bladder cancer cell line T24 (TGR) by quantitative real-time PCR (qRT-PCR). Results360 genes were differentially expressed between BLCA and normal bladder mucosae simultaneously in three GEO datasets, of which 59 were up-regulated and 301 were down-regulated. 159 prognostic genes were obtained from DEGs. Lasso and multivariate cox regression were conducted in sequence and the prognostic model was eventually optimized to four genes (EHBP1, RHOJ, FASN, STXBP6). Survival analyses demonstrated that the overall survival (OS) of patients in high-risk group was significantly shorter than that in low-risk group. The area under the curve (AUC) values of 3–5 years survival were basically above 0.7. Moreover, cox regression analyses showed that age, T stage and risk score were independent indicators for BLCA prognosis. For further clinical application, a nomogram was then constructed by integrating these factors. The C-index (0.72, CI 95 %, 0.669–0.775) and calibration curves demonstrated the good performance of nomogram. Importantly, the mRNA level of model genes was significantly up-regulated in TGR compared to T24, indicating a better prediction for chemotherapy-resistant BLCA patients. ConclusionCollectively, our findings suggest a novel four-gene predictive model for BLCA prognosis. It is expected to provide a valuable reference for prognostic evaluation and treatment in BLCA patients.

The value of ATAD3A as a potential biomarker for bladder cancer.

Bladder cancer (BCa) is a highly malignant tumor, and if left untreated, it can develop severe hematuria and tumor metastasis, thereby endangering the patient's life. The purpose of this paper was to detect the expression of ATAD3A in BCa and research the relationship between ATAD3A and pathological features of bladder cancer and the prognosis of patients. First, the expression of ATAD3A in BCa and normal bladder tissues was analyzed based on the UALCAN and Oncomine public databases. Second, 491 cases of surgically resected bladder cancer specimens and 110 cases of normal adjacent tissues were immunohistochemically stained. The expression of ATAD3A was quantified, and the value and prognosis of ATAD3A as a biomarker of BCa were evaluated. The expression of ATAD3A in bladder cancer tissues was higher than that in normal bladder mucosa. High expression of ATAD3A was correlated with patient age, tumor size, number of tumors, distant metastasis, lymph node metastasis, lymphovascular invasion, and TNM stage (p < 0.05). Overexpression of ATAD3A is closely related to cancer patient survival. The mean survival time of bladder cancer patients with high ATAD3A expression was shorter than those with low ATAD3A levels. According to the relative comparing result, the high ATAD3A expression herald reduced overall survival in BCa patients. The abnormal overexpression of ATAD3A may be related to the initiation and progress of bladder cancer. The upregulation of ATAD3A can be used as an effective indicator to diagnose bladder cancer and predict tumor progression. Furthermore, the combination of information from public databases and the results of clinical sample analysis can help us better understand the mechanism of action of molecular oncogenes in bladder cancer.

The classification of the bladder cancer based on Vision Transformers (ViT)

Bladder cancer is a prevalent malignancy with diverse subtypes, including invasive and non-invasive tissue. Accurate classification of these subtypes is crucial for personalized treatment and prognosis. In this paper, we present a comprehensive study on the classification of bladder cancer into into three classes, two of them are the malignant set as non invasive type and invasive type and one set is the normal bladder mucosa to be used as stander measurement for computer deep learning. We utilized a dataset containing histopathological images of bladder tissue samples, split into a training set (70%), a validation set (15%), and a test set (15%). Four different deep-learning architectures were evaluated for their performance in classifying bladder cancer, EfficientNetB2, InceptionResNetV2, InceptionV3, and ResNet50V2. Additionally, we explored the potential of Vision Transformers with two different configurations, ViT_B32 and ViT_B16, for this classification task. Our experimental results revealed significant variations in the models’ accuracies for classifying bladder cancer. The highest accuracy was achieved using the InceptionResNetV2 model, with an impressive accuracy of 98.73%. Vision Transformers also showed promising results, with ViT_B32 achieving an accuracy of 99.49%, and ViT_B16 achieving an accuracy of 99.23%. EfficientNetB2 and ResNet50V2 also exhibited competitive performances, achieving accuracies of 95.43% and 93%, respectively. In conclusion, our study demonstrates that deep learning models, particularly Vision Transformers (ViT_B32 and ViT_B16), can effectively classify bladder cancer into its three classes with high accuracy. These findings have potential implications for aiding clinical decision-making and improving patient outcomes in the field of oncology.

MRNA expression of CRF family members in urothelial bladder cancer.

The corticotropin-releasing factor (CRF) gene family includes the three urocortins (UCN1, 2 and 3) and the two receptors (CRFR1 and 2), which play a significant role in the physiology of various organs. The expression of the CRF family of genes and its receptors are shown to participate in the pathogenesis of inflammation and even tumorigenesis. However, data regarding the human urinary tract, especially the bladder, are scarce. To the best of our knowledge, no studies are currently available on the CRF system and bladder cancer. The primary goal of the present study was to investigate the mRNA expression of the CRF family members in bladder cancer. The secondary aim was to analyze the differences with the expression of the same mRNAs in normal bladders. From August 2018 to July 2021, 43 recruited patients were divided into three groups. Group A included healthy patients, group B included patients with bladder cancer and group C included patients with a history of cancer from whom samples were taken from the normal bladder mucosa. Detection of mRNA of the CRF family of genes was performed using reverse transcription-quantitative PCR. The mRNA of the three urocortins, CRF and the two receptors were predominantly expressed in all three groups of patients. Statistical analysis using the Kruskal-Wallis test showed that UCN1 was downregulated in patients with bladder cancer and those with possible cancer compared with the healthy group (mean rank group A=24.3 vs. mean rank group B=12.58; P=0.006) and (mean rank group A=24.3 vs. mean rank group C=8.88; P=0.001). The present experiments showed that mRNA of the CRF family of genes was amplified in normal and cancer bladder tissues. Downregulation of the UCN1 gene may be associated with bladder cancer, contributing to the prognosis, diagnosis or therapy of urothelial malignancies.

TGFB Induced Factor Homeobox 2 Induces Deterioration of Bladder Carcinoma via Activating CD2 Cytoplasmic Tail Binding Protein 2

Bladder carcinoma is a complex and aggressive malignancy with limited treatment options. In this study, we aimed to investigate the expression pattern of TGIF2 in bladder carcinoma and its clinical significance, as well as its functional role and interaction with CD2BP2 in disease progression. Through quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis, we found that TGIF2 was highly expressed in bladder carcinoma tissues compared to normal bladder mucosa. Furthermore, elevated TGIF2 levels were associated with advanced tumor stage and larger tumor size, indicating its potential as a prognostic marker in bladder carcinoma. Using knockdown models in bladder carcinoma cell lines (253j and J82), we observed that the inhibition of TGIF2 resulted in decreased proliferation and migration rates, suggesting a critical role of TGIF2 in promoting these malignant phenotypes. Additionally, our dual-luciferase reporter assay revealed a direct interaction between TGIF2 and CD2BP2, with CD2BP2 being upregulated in bladder carcinoma tissues and positively correlated with TGIF2 expression. Notably, the overexpression of CD2BP2 reversed the suppressed malignant phenotypes caused by TGIF2 knockdown. Collectively, our findings highlight the abundant expression of TGIF2 in bladder carcinoma tissues and its association with malignant characteristics. We demonstrate that TGIF2 promotes proliferative and metastatic capacities in bladder carcinoma by positively regulating CD2BP2. These insights provide a basis for further investigations into the potential of TGIF2 and CD2BP2 as therapeutic targets and prognostic markers in bladder carcinoma management.

Efficacy of allogeneic platelet growth factors in actinic cystitis: The resolution of trouble?

BackgroundActinic cystitis is a severe complication after radiotherapy for prostate cancer. It is a chronic inflammatory process that leads to an alteration of bladder mucosa with formation of petechiae and subsequently hematuria. Actinic cystitis responds poorly to medical treatment, with a heavy burden on patients’ quality of life. Patients with refractory hematuria may undergo cystectomy in the attempt to control bleeding. We conducted a prospective study to evaluate the effectiveness of the allogeneic platelet growth factors for actinic cystitis. Methods and MaterialNine patients with actinic cystitis were enrolled in this study. The primary outcome measures were the effects of the platelet growth factors on the injury of the bladder mucosa. The secondary outcome was the change in quality of life ResultsA total of 9 patients, mean age 68 (range 59–81) underwent a therapeutic program of bladder instillation with allogeneic platelets growth factors for 3 months. Of the 9 patients, all (100 %) had complete resolution of hematuria and urinary symptoms. After three months cystoscopy showed regeneration of the normal bladder mucosa. Biopsies allowed histological confirmation of the finding. DiscussionThe instillation of allogeneic platelet growth factors in actinic cystitis is a new treatment that in this setting of patients appears promising in promoting a resolution of urinary symptoms, hematuria and avoiding a disabling surgery such as cystectomy.

Single-cell RNA sequencing reveals the immune microenvironment and signaling networks in cystitis glandularis.

Cystitis glandularis (CG) is a rare chronic bladder hyperplastic disease that mainly manifests by recurrent frequent urination, dysuria and gross hematuria. The current lack of unified diagnosis and treatment criteria makes it essential to comprehensively describe the inflammatory immune environment in CG research. Here, we performed scRNA-sequencing in CG patients for the first time, in which four inflamed tissues as well as three surrounding normal bladder mucosa tissues were included. Specifically, we isolated 18,869 cells to conduct bioinformatic analysis and performed immunofluorescence experiments. Our genetic results demonstrate that CG does not have the classic chromosomal variation observed in bladder tumors, reveal the specific effects of TNF in KRT15 epithelial cells, and identify a new population of PIGR epithelial cells with high immunogenicity. In addition, we confirmed the activation difference of various kinds of T cells during chronic bladder inflammation and discovered a new group of CD27-Switch memory B cells expressing a variety of immunoglobulins. CG was regarded as a rare disease and its basic study is still weak.Our study reveals, for the first time, the different kinds of cell subgroups in CG and provides the necessary basis for the clinical treatment of cystitis glandularis. Besides, our study significantly advances the research on cystitis glandularis at the cellular level and provides a theoretical basis for the future treatment of cystitis glandularis.

Clinical significance of galectin-3 expression in urinary bladder carcinoma

To uncover the clinical significance of galectin-3 in the evolution of urinary bladder cancer by defining galectin-3 expression and examining the relationship between its expression in a group of urothelial carcinomas versus normal tissues along with clinicopathological factors. This retrospective study included histopathological reports and archival blocks and slides of all patients with urinary bladder cancer treated at King Abdulaziz University Hospital (Jeddah, Saudi Arabia). An anti-galectin-3 monoclonal antibody was used for immunohistochemical staining of tissue microarray slides comprising 128 cases of urothelium carcinoma and 24 specimens of normal bladder mucosa. Galectin-3 was downregulated during transformation, with positive expression found in 50 (39%) urinary bladder neoplasms, of which 33 (66%) showed weak immunostaining. All positively-stained malignant tumor and normal bladder mucosa samples showed cytoplasmic staining; a few samples also showed nuclear staining. No correlation was noted between galectin-3 and histotype, grade, stage, muscularis propria invasion, lymph node invasion, vascular invasion, or metastasis. A Cox proportional hazards model and Kaplan-Meier survival curves did not show differences in survival on the basis of galectin-3 expression. Galectin-3 is down-regulated in bladder cancer but is not a helpful marker for the diagnosis or prognosis of urinary bladder cancer.

Acquired childhood bladder melanosis

BackgroundBladder melanosis is rare. It has previously been described only in the fifth decade of life or beyond; it has not been described in childhood. Previous descriptions have generally been case reports, and so the natural history is poorly understood. Urinary symptoms present at the time of cystoscopy have frequently been attributed to bladder melanosis. A possible suggested aetiology is aberrant migration of melanocyte migration during embryogenesis.Case presentationWe present the first case of bladder melanosis in a child. He had been under our care since the age of 5 years with urinary incontinence and at that time, had undergone cystoscopy demonstrating normal bladder mucosa. A diagnosis of idiopathic detrusor overactivity with underactive voiding had been made. After other unsuccessful treatments, intravesical botulinum toxin was proposed. At the age of 13, repeat cystoscopy prior to botulinum toxin, demonstrated widespread pigmented areas in the bladder mucosa. Histology showed bladder melanosis. Our finding is important for several reasons. This is the first reported case of bladder melanosis to affect a child. The previous normal cystoscopy in our patient would refute the explanation that bladder melanosis is a congenital condition. Furthermore, the development of melanosis on the background of stable symptoms raises the possibility that the condition may be asymptomatic.ConclusionsThis unique finding of bladder melanosis in a child has provided further insight into this rare and poorly understood condition.

MP59-13 BLADDER TUMOR ACCURATE RADIOFREQUENCY RESECTION: 38 CASES REPORTS

You have accessJournal of UrologyCME1 May 2022MP59-13 BLADDER TUMOR ACCURATE RADIOFREQUENCY RESECTION: 38 CASES REPORTS Shengkun Sun, Guangfu Chen, Weijun Fu, and Xu Zhang Shengkun SunShengkun Sun More articles by this author , Guangfu ChenGuangfu Chen More articles by this author , Weijun FuWeijun Fu More articles by this author , and Xu ZhangXu Zhang More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002642.13AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Bladder cancer is the fourth most frequently diagnosed malignancy in males and the 12th most common malignancy in females. Transurethral resection of bladder tumor (TURBT) remains the mainstay of diagnosis and initial treatment of bladder neoplasms. TURBT is performed conventionally with an electrocautery loop. The piecemeal resection specimens are inappropriate for tumor pathological evaluation because of poor anatomic orientation and evaluation of the pathological depth is not necessarily accurate. Also, tumor cells are scattered as the tumor is fragmented so that the possibility of implantation cannot be excluded. The objective of this study is to optimize the surgical method of transurethral bladder tumor resection. METHODS: 38 patients with non-muscle invasive bladder cancer received transurethral accurate resection of the tumor with a novel β electrode with a radiofrequency energy platform. Normal bladder mucosa 0.5 to 1 cm from the tumor base was marked with the tip of β electrode with an electrocautery energy. The needle of the β electrode was insert into the submucosa, and tissue of the bladder wall was pulled towards the midline of the bladder, followed by electro-cutting. Tumor blood vessels were cauterized with the needle tip. After en bloc resection of the tumor bulk, a novel β basket, which could be opened and trapped the tumor in bladder, was introduced into the bladder to retrieve the tumor en bloc or sub-en bloc. RESULTS: Tumors exceeded T2 stage or tumors with a maximum diameter of less than 1 cm were excluded. 38 patients underwent accurate transurethral radiofrequency resection of bladder tumors, and a total of 47 tumors were removed. The maximum diameter of the tumor is 2.4cm (range from 1 to 5cm). Tumors less than 3cm could be retrieved through the resection sheath or be taken out with the β basket. 12 tumors with a maximum diameter of >3cm were divided into 2-3 large pieces by the β basket and then taken out. During the operation, there was no obturator nerve reflex, no bladder perforation or urine extravasation. The intraoperative bleeding was less than 2ml in every patient. The bladder was flushed with normal saline for 24 hours post operation. Postoperative pathology showed Ta in 27cases and T1 in 11 cases. No second TURBT was performed in this group. No operative complications occurred. All patients received intravesical irrigation of BCG or chemotherapy agent periodically. 2 patients had tumor recurrence in the bladder during the mean follow-up of 8.7 months (range from 3 to 11 months). CONCLUSIONS: A novel β electrode was used to remove the tumor during accurate transurethral radiofrequency resection of bladder tumors. There are several advantages of this novel method. First, the blood supply of the tumor was occluded to avoid bleeding. Second, the obturator nerve reflex could be prevented completely and local nerve blockage was not necessary. Third, accurate radiofrequency resection of bladder tumor improved the resection quality and the en bloc resection helped make accurate TNM stage which was significant for postoperative treatment strategy. Source of Funding: None © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e1007 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Shengkun Sun More articles by this author Guangfu Chen More articles by this author Weijun Fu More articles by this author Xu Zhang More articles by this author Expand All Advertisement PDF DownloadLoading ...

No Added Benefit of Systematic Bladder Biopsies after BCG Induction in High-Risk Non-Muscle-Invasive Bladder Cancer Optimally Managed

Objectives: To evaluate the added detection benefit of routine transurethral biopsies after an induction course of intravesical Bacillus Calmette-Guérin (BCG) for high-grade NMIBC. Methods: We retrospectively included all patients who underwent white and blue light TURB, subsequent adjuvant induction scheme of BCG, followed by post-instillation biopsies. We excluded patients in whom a prior BCG induction course failed those with synchronous upper tract urothelial carcinoma, and those who did not receive the full 6-week course. A total of 62 patients met inclusion criteria and their records were analyzed. Cystoscopy findings, urine cytology and pathological results of transurethral biopsy were evaluated. Sensitivity, specificity, positive predictive value, negative predictive value, positive and negative likelihood ratios and diagnostic accuracy were calculated for cystoscopy and cytology separately and in combination. Results: After BCG, a total of 342 bladder biopsy were taken with a median of 5 (IQR 2) per case. The biopsy was positive in 13 patients (21%). The median number of positive biopsies was 2 (IQR 1) and the total number of positive biopsies was 27 (8%). The biopsy was positive at the initial tumor site in 8 patients (61.5%). No complications of Clavien III to V were reported. Out of 62 patients who underwent systematic transurethral biopsy, only 3 (5%) presented complications from grade I to II. We found a significant association of positive biopsy with positive urinary cytology (p &lt;0.001) and the presence of erythematous lesions (p &lt;0.02) or visible tumors (p &lt;0.001) at cystoscopy. The pre-BCG stage T (p = 0.32), the presence of CIS (p = 0.71), the multifocal tumor (p = 0.41) and the size (p = 0.52) were not associated with a positive biopsy occurrence. For normal bladder mucosa the predictive value of negative biopsy in cases with negative vs positive cytology was 98% vs 79% (p 0.001). Conclusion: Routine transurethral bladder biopsies after a BCG induction course are not mandatory in the modern era of NMIBC management.

MP10-03 SUPER TRANSURETHRAL BLADDER TUMOR RESECTION

You have accessJournal of UrologySurgical Technology & Simulation: Instrumentation & Technology I (MP10)1 Sep 2021MP10-03 SUPER TRANSURETHRAL BLADDER TUMOR RESECTION Shengkun Sun, Yong Xu, Weijun Fu, Guangfu Chen, and Xu Zhang Shengkun SunShengkun Sun More articles by this author , Yong XuYong Xu More articles by this author , Weijun FuWeijun Fu More articles by this author , Guangfu ChenGuangfu Chen More articles by this author , and Xu ZhangXu Zhang More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000001983.03AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: The existing TURBT technology has a history of more than 80 years. The fragmented resection of tumors are inherent defects of traditional TURBT. The objective of this study was to explore a safe and effective method of TURBT. We performed a novel transurethral bladder tumor resection under air cystoscope. Since the depth of resection is limitless under pneumocystoscope, and there is no previous report, we named this novel operation as “Super Transurethral Resection of Bladder Tumor” (STURBT). METHODS: The operation was performed under general or epidural anesthesia. The Super Scope (S-Scope) was inserted through the urethra, and the bladder was filled with carbon dioxide, and the pressure was set as 12 mmHg. The novel S-Scope has a channel to accommodate conventional laparoscopic surgical instruments, such as supersonic scalpel, bipolar scissors, forceps, and suctions. First the normal bladder mucosa 1 cm from the tumor was marked with a supersonic scalpel. Blunt separation and sharp resection were performed from periphery to the central base of the tumor to achieve en bloc resection. The β basket, a bladder tumor retrieving device, was inserted through the channel, and the tumor was removed in whole or by dividing into 2 to 3 parts. The urethral stent was placed, and the bladder was flushed with normal saline. RESULTS: 5 patients underwent STURBT, and 6 tumors were resected with the largest diameter of 3∼5cm. The average resection time was 28±16.4 minutes, and the tumor was retrieved en bloc or sub-en bloc. There was no obturator nerve reflex or bleeding during operation. Pathological diagnosis showed 1 case of pheochromocytoma and 4 cases of high grade urothelial carcinoma with stage T1∼T2a. Bladder muscle layer was visible in all cases. 3 cases of bladder cancer with muscle invasion received adjuvant intravenous chemotherapy. 4 cases with high grade urothelial carcinoma also received intravesical irragation of BCG periodically. Cystoscopy 3 months post-operation showed well healing of the bladder. There was no tumor recurrence during a mean follow-up of 11.8 (3-18) months. CONCLUSIONS: STURBT is a novel surgical method to treat bladder tumor, which has several advantages over traditional TURBT. No fluid irrigation was needed, and no urine extravasation developed. The supersonic scalpel will not stimulate the obturator nerve. The en bloc tumor resection decreased tumor dissemination and an accurate pathological TNM stage could be obtained. S-Scope assisted STURBT could make full use of the current laparoscopic surgical instruments, which had obvious advantages in transurethral treatment of bladder tumors. Source of Funding: None © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e169-e169 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Shengkun Sun More articles by this author Yong Xu More articles by this author Weijun Fu More articles by this author Guangfu Chen More articles by this author Xu Zhang More articles by this author Expand All Advertisement Loading ...

Integrative Analysis Identified MCT4 as an Independent Prognostic Factor for Bladder Cancer.

BackgroundBladder cancer is the 10th most common cancer and most common urothelial malignancy worldwide. Prognostic biomarkers for bladder cancer patients are required for individualized treatment. Monocarboxylate transporter 4 (MCT4), encoded by SLC16A3 gene, is a potential biomarker for bladder cancer because of its crucial role in the lactate efflux in the aerobic glycolysis process. We aimed to study the association between MCT4 expression and the overall survival (OS) of bladder cancer patients.MethodsThe published single-cell RNA sequencing data of 49,869 bladder cancer cells and 15,827 normal bladder mucosa cells and The Cancer Genome Atlas (TCGA) bladder cancer cohort data were used to explore the mRNA expression of SLC16A3 in bladder cancer. Eighty-nine consecutive bladder cancer patients who had undergone radical cystectomy were enrolled as a validation cohort. The expression of MCT4 proteins in bladder cancer specimens was detected using immunohistochemistry staining. The Kaplan–Meier survival analysis and Cox regression were performed to analyze the association between MCT4 protein expression and OS in bladder cancer patients.ResultsSLC16A3 mRNA was upregulated in bladder cancer cells. The upregulated genes in SLC16A3-positive epithelial cells were enriched in the glycolysis process pathway and monocarboxylic acid metabolic process pathway. Patients with high SLC16A3 mRNA expression showed significantly poor OS (p = 0.016). High MCT4 protein expression was also found to be an independent predictor for poor OS in bladder cancer patients (HR: 2.462; 95% CI: 1.202~5.042, p = 0.014). A nomogram was built based on the results of the multivariate Cox analysis.ConclusionBladder cancer with high SLC16A3 mRNA expression has a poor OS. High MCT4 protein expression is an independent prognostic factor for bladder cancer patients who had undergone radical cystectomy.

Gas6 expression and Tyrosine kinase Axl Sky receptors: Their relation with tumor stage and grade in patients with bladder cancer.

It has been shown that the dysregulation of tyrosine kinase Axl receptor and its ligand growth arrest-specific gene (Gas6) are associated with poor prognosis in various types of tumors but there is not enough study about their importance in bladder cancer (BC). We evaluated the relation of Gas6 gene expression and tyrosine- kinase Axl and Sky (Tyro 3) receptors with tumor stage and grade in patients with BC. The study group consists of 55 patients whose transurethral resection of bladder (TUR-B) has been performed due to BC and the control group consists of 12 patients with normal bladder mucosa. In tissues mRNAs of Gas6, Axl, and Sky receptors were examined by quantitative (Real-Time) PCR (qPCR). Protein expression was measured by immunohistochemistry. Plasma Gas6 protein levels were compared with control group by ELISA method. Patients with BC were grouped as Ta low (n=17), Ta high (n=5), T1 low (n=9), T1 high (n=8) and T2 (n=16) according to their TUR-B pathologies. The qPCR analysis showed that the expression of Gas6 gene and Axl receptor is higher in the tumor-positive group and the immune-histochemical showed that the bladder samples of the tumor-positive group stained significantly positive. When the patients are grouped according to the TUR-B pathologies, a statistical significant difference was observed among groups in the qPCR analysis ratios of Gas6 gene and Axl receptor by (p < 0.05) but no significance was found for Sky receptor (p > 0.05). When Gas6 protein levels in plasma samples were compared by ELISA method, a statistical significance was determined among groups (p = 0.001). Our findings indicate that mRNAs of Gas6 and Axl receptor are closely related to tumor stage and grade in patients with BC. Further studies are needed for understanding the role of Gas6 and its receptors on the neoplastic transformation in terms of novel biomarkers and potential therapeutic targets.

Mullerianosis of the urinary bladder; A case report.

IntroductionMullerianosis is a sporadic condition of the urinary bladder. The study aims to present a case of mullerianosis with a brief literature review.Case presentationA 52-year-old lady presented with hematuria for one-year duration. A general urine examination showed microscopic hematuria. Abdominal and transvaginal ultrasonography revealed a soft tissue mass (13 * 10 mm) within the base of the urinary bladder. Cystoscopy showed 1.0 * 1.3 cm benign-looking mass bulging into the posterior bladder wall covered by a normal bladder mucosa. Transurethral resection was done. The histopathological examination was consistent with mullerianosis.DiscussionTwo hypotheses have been proposed; the first one (implantation theory) believing that at the time of pelvic surgery and caesarian delivery Mullerian tissues become implanted inside the bladder. The second theory (metaplastic theory) holds the probability of differentiation of Mullerian epithelium to endometrial, endocervical, and tubal components and proliferation in the bladder.Conclusionmullerianosis is a very infrequent entity that mainly affects the urinary bladder and is mainly reported in females during the reproductive period, medical and surgical treatment are both regarded as the management options.

Detection and Quantification of the E6 Oncogene in Bovine Papillomavirus Types 2 and 13 From Urinary Bladder Lesions of Cattle.

Bovine papillomavirus types 2 and 13 can induce tumors in both the cutaneous and mucosal epithelia of cattle. These viral types are associated with the development of benign cutaneous papillomas and malignant lesions in the urinary bladders of cattle, with the latter being known as bovine enzootic hematuria. Among the viral oncoproteins encoded by Deltapapillomavirus DNA, the E6 oncoprotein has an important role in cell proliferation and might be related to cancer initiation and promotion. The aim of this study was to present a standardized SYBR Green-based quantitative PCR for detection and quantification of the bovine papillomavirus 2 and 13 E6 oncogenes in urinary bladder samples from cattle. Twenty-four urinary bladders from cattle displaying tumors (n = 12) and normal bladder mucosa (n = 12) were tested by quantitative PCR. Of the 12 urinary bladders with tumors, six presented bovine papillomavirus 2 DNA concentrations ranging from 1.05 × 104 to 9.53 × 103 copies/μL, while two had bovine papillomavirus 13 DNA amplified at concentrations of 1.30 × 104 to 1.23 × 104 copies/μL. The healthy bladder mucosa samples were negative for both bovine papillomaviruses. Once the results were confirmed by conventional PCR and direct sequencing, the quantitative PCR assay developed in this study was shown to be a sensitive and specific tool for detecting and quantifying the E6 ORF of bovine papillomavirus 2 and 13 in a variety of clinical samples. Our findings of identification of bovine papillomavirus 2 and 13 DNA in urothelial tumors from cattle suffering from bovine enzootic hematuria agree with data from previous studies, representing the first detection of bovine papillomavirus 13 DNA in malignant bladder lesions of cattle from Brazil.

Identification of potential genes in upper tract urothelial carcinoma using next-generation sequencing with bioinformatics and in vitro analyses.

BackgroundWe aimed to identify prognostic biomarkers of upper tract urothelial carcinomas (UTUCs), including microRNAs (miRNAs) and genes which account for only 5% to 10% of all urothelial carcinomas (UCs). In Taiwan, this figure is markedly higher, where it can reach up to 30% of UC cases.Materials and MethodsUsing next-generation sequencing (NGS), we analyzed two pairs of renal pelvis tumors and adjacent normal urothelial tissues to screen miRNAs and messenger RNAs. By combining bioinformatics analysis from miRmap, Gene Expression Omnibus (GEO), and Oncomine and Ingenuity® Pathway Analysis databases, we identified candidate genes. To search for upstream miRNAs with exact target binding sites, we used miRmap, TargetScan, and miRDB to enforce evidence. Then, we clarified gene and protein expression through an in vitro study using western blot analysis and quantitative real-time reverse transcriptase-PCR.ResultsInteractions between selected target genes obtained using the NGS and miRmap methods were assessed through a Venn diagram analysis. Six potential genes, namely, PDE5A, RECK, ZEB2, NCALD, PLCXD3 and CYBRD1 showed significant differences. Further analysis of gene expression from the GEO dataset indicated lower expression of PDE5A, RECK, ZEB2, and CYBRD1 in bladder cancer tissue than in normal bladder mucosa, which indicated that PDE5A, RECK, ZEB2, and CYBRD1 may act as tumor suppressors in UTUC. In addition, we compared the expression of these genes in various UC cell lines (RT4, BFTC905, J82, T24, UMUC3, 5637, BFTC 909, UMUC14) and found decreased expression of PDE5A in muscle-invasive UC cells compared with the RT4 cell line. Furthermore, by using paired UTUC and normal tissues from 20 patients, lower PDE5A expression was also demonstrated in tumor specimens.ConclusionsOur findings suggest these candidate genes may play some roles in UTUC progression. We propose that these markers may be potential targets clarified by in vitro and in vivo experiments. PDE5A also potentially presents tumor suppressor genes, as identified by comparing the expression between normal and tumor specimens.

Differential Expression Analysis for the Identification of Survival Associated Genes in Primary Bladder Cancer using Microarray Data

Bladder cancer (BC) is a highly malignant and malevolent tumor whose incidence and mortality worldwide remains high. This paper aimed to identify the differentially expressed genes (DEGs) between normal and primary bladder cancer samples, which might be of prognostic value, and evaluate their association to clinical outcomes (survival) in patients with primary bladder cancer. A sample of 256 gene expression profiles of which 10 were normal bladder mucosae whereas 165 were primary bladder cancer tissues were downloaded from the gene expression omnibus (GEO). The limma package in R was used to perform screening for DEGs after variance reduction through quantile normalization and log2 transformation. Gene Ontology (GO), KEGG enrichment analysis, and Network analysis were performed to determine the functions of these DEGs. Cox proportional hazard regression model was then used to determine the association between the top selected DEGs and overall primary bladder cancer survival. Selected genes were then encoded as either low, middle or high expression and Kaplan-Meier plots then used to visualize the effect of these expression levels for the selected five genes on bladder cancer survival probability. This study identified a total of 5, 979 DEGs where 2, 878 were upregulated, 3, 101 were downregulated and 37, 169 were not significantly expressed differently. Cox proportional hazard stepwise regression models revealed a number of DEGs that have a statistically significant association to bladder cancer. The selected five DEGs associated with survival were EDNRB (HR=1.9858, 95%CI 1.4965–2.6350), COL14A1 (HR=1.5335, 95%C1 1.1670–2.0151), MOXD1 (HR=1.4878, 95%CI 1.1070–1.9996), FAM107A (HR=1.4970, 95%CI 1.1162–2.0077), and REM1 (HR=1.5999, 95%CI 1.2422–2.0605). After encoding these genes as either low, middle, or high expression, high expression of both EDNRB, COL14A1, FAM107A, and REM1 were found to result into statistically significant reductions in survival probabilities of patients (p0.05). The genes found in this study could provide useful insight in the understanding of the association between the identified DEGs and primary bladder cancer survival, which can further inform future studies and clinical therapies.

C1QTNF6 Overexpression Acts as a Predictor of Poor Prognosis in Bladder Cancer Patients.

Background Bladder cancer is one of the most common urinary malignancies. This study is aimed at providing some promising molecular biomarkers for bladder cancer (BC) by investigating the correlation between C1QTNF6 expression and clinical characteristics as well as prognosis in patients with bladder cancer. Methods Sequencing profiles of C1QTNF6 mRNA in BC patients were collected to evaluate the distinctive gene expression, between normal bladder mucosa and BC, according to the TCGA and GEO databases. The association between C1QTNF6 expression and the clinical features as well as the disease prognosis was evaluated using two independent cohorts. The expression of C1QTNF6 in normal bladder and BC cells was examined by western blotting and PCR, so the underlying molecular mechanism could be further investigated. Results C1QTNF6 mRNA levels were found to be differentially expressed in two independent public cohorts, including the TCGA database and GSE13507 dataset from GEO. The protein and RNA levels of C1QTNF6 in BC cells were both elevated when compared to normal bladder cell lines. High C1QTNF6 expression was detected in advanced T/M stages, pathological grade, and AJCC stage when compared to the low C1QTNF6 expression group. The underlying mechanism related to this differential expression could be explained by cell migration and invasion assays, where bladder cancer cells 5637 and T24 had a significant reduction on migration and invasion ability upon knockdown of C1QTNF6 expression. The low C1QTNF6 expression group presented a more prominent OS advantage over the high-expression group in both TCGA and GSE13507 cohorts. Moreover, the protein content in tissues was further validated using the HPA database and TMA. Survival analyses also indicated that the high C1QTNF6 expression group had an unfavorable OS when compared to the low-expression group. Conclusions High C1QTNF6 expression may serve as a predictor of poor prognosis in bladder cancer patients, and the underlying mechanism is possibly associated with changes on cancer cell migration and invasion ability.

Abstract PR05: Similarities in molecular subtypes and subtype immune patterns between naturally occurring canine and human invasive bladder cancer

Abstract Relevant animal models are needed to study treatment strategies related to the molecular subtypes of bladder cancer and the immune response between these subtypes. The purpose of the study was to characterize the molecular subtypes and immune features in naturally occurring canine invasive urothelial carcinoma (InvUC). Canine InvUC is already known to mimic the human condition in pathologic features, metastatic behavior, and chemotherapy response. Molecular subtyping could further increase the utilization of the canine model. RNA-seq data from canine InvUC (n=56) and normal canine bladder mucosa (n=4) were analyzed using Strand NGS (Strand Genomics, San Francisco, CA). Data were normalized (DESeq, TMM), and DEGs selected (2FC, pcorr &amp;lt;0.05) using DESeq2 and edge R, respectively. Following unsupervised clustering that identified two groups of tumors, supervised clustering was performed in which the data were interrogated using a panel of genes known to classify human bladder cancer subtypes (TCGA), as well as multiple gene panels that classify human bladder cancer as immune hot (infiltrated) or immune cold (noninfiltrated, immune-suppressor features). Infiltrating immune cells were observed in histologic sections with CD3 immunohistochemistry. Distinct luminal (n=28) and basal (n=28) subtypes were identified in the canine InvUC samples, with subclassifications emerging in both subtypes. There was anticipated heterogeneity between the samples, but of the 28 basal tumors, all had hot immune features, with 10 being exceptionally hot. Of the luminal tumors, 22 of 28 were notably immune cold. In conclusion, canine InvUC mimics human invasive bladder cancer in regard to molecular subtypes and immune patterns within subtypes. This expands the value of the canine bladder cancer model and offers opportunities to study strategies for individualized therapy and to develop strategies to enhance the tumor immune environment in order to improve treatment outcomes. Support: NIH/NCI P30CA023168 Supp (Knapp, Ratliff). This abstract is also being presented as Poster A26. Citation Format: Deepika Dhawan, Breann C. Sommer, José A. Ramos-Vara, Noah M. Hahn, Deborah W. Knapp. Similarities in molecular subtypes and subtype immune patterns between naturally occurring canine and human invasive bladder cancer [abstract]. In: Proceedings of the AACR Special Conference on Bladder Cancer: Transforming the Field; 2019 May 18-21; Denver, CO. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(15_Suppl):Abstract nr PR05.