Non Invasive Bladder Cancer Research Articles

Machine Learning-Based Detection of Bladder Cancer by Urine cfDNA Fragmentation Hotspots that Capture Cancer-Associated Molecular Features.

cfDNA fragmentomics-based liquid biopsy is a potential option for noninvasive bladder cancer (BLCA) detection that remains an unmet clinical need. We assessed the diagnostic performance of cfDNA hotspot-driven machine-learning models in a cohort of 55 BLCA patients, 51 subjects with benign conditions, and 11 healthy volunteers. We further performed functional bioinformatics analysis for biological understanding and interpretation of the tool's diagnostic capability. Urinary cfDNA hotspots-based machine-learning model enabled effective BLCA detection, achieving high performance (area under curve 0.96) and an 87% sensitivity at 100% specificity. It outperformed models using other cfDNA-derived features. In stage-stratified analysis, the sensitivity at 100% specificity of the urine hotspots-based model was 71% and 92% for early (low-grade Ta and T1) and advanced (high-grade T1 and muscle-invasive) disease, respectively. Biologically, cfDNA hotspots effectively retrieved regulatory elements and were correlated with the cell of origin. Urine cfDNA hotspots specifically captured BLCA-related molecular features, including key functional pathways, chromosome loci associated with BLCA risk as identified in genome-wide association studies, or presenting frequent somatic alterations in BLCA tumors, and the transcription factor regulatory landscape. Our findings support the applicability of urine cfDNA fragmentation hotspots for noninvasive BLCA diagnosis, as well as for future translational study regarding its molecular pathology and heterogeneity.

Noninvasive Bladder Cancer Treated with Bacillus Calmette-Guérin in Kidney Transplant Recipients

Noninvasive Bladder Cancer Treated with Bacillus Calmette-Guérin in Kidney Transplant Recipients

Nanomedicine in Bladder Cancer Therapy.

Bladder cancer (BC) is one of the most common malignant neoplasms of the genitourinary system. Traditional BC therapies include chemotherapy, targeted therapy, and immunotherapy. However, limitations such as lack of specificity, cytotoxicity, and multidrug resistance pose serious challenges to the benefits of BC therapies. Consequently, current studies focus on the search for new therapeutic solutions. In recent years, there has been a growing interest in using nanotechnology in the treatment of both non-invasive (NMIBC) and invasive bladder cancer (MIBC). Nanotechnology is based on the use of both organic molecules (chitosan, liposomes) and inorganic molecules (superparamagnetic iron oxide nanoparticles) as carriers of active substances. The main aim of such molecules is the targeted transport and prolonged retention of the drug in the target tissue, which increases the therapeutic efficacy of the active substance. This review discusses the numerous types of nanoparticles (including chitosan, polymeric nanoparticles, liposomes, and protein nanoparticles), targeting mechanisms, and approved nanotherapeutics with oncological implications in cancer treatment. We also present nanoformulation applications in phototherapy, gene therapy, and immunotherapy. Moreover, we summarise the current perspectives, advantages, and challenges in clinical translation.

Comparison of Tissue and Urine Microbiota in Male, Intervention Naive Patients with and without Non-Invasive Bladder Cancer

<bold></bold> Introduction: To investigate the presence of dysbiosis in patients with naive bladder cancer. Methods: Twelve male patients with non-invasive bladder cancer and twelve age-matched healthy males had midstream urine and tissue samples taken. A history of endourological interventions was determined as an exclusion criterion, ensuring that the study was designed solely with naïve participants. The bacterial 16s ribosomal RNA V3-V4 regions were used to examine urine and tissue samples. We compared the microbiota composition of the bladder cancer and control groups. Results: Escherichia Shigella (p < 0.001), Staphylococcus (p < 0.001), Delftia (p < 0.001), Acinetobacter (p < 0.001), Corynebacterium (p < 0.001), and Enhydrobacter (p < 0.001) were abundant in bladder cancer tissue samples. Escherichia Shigella (p < 0.001), Ureaplasma (p < 0.001), Lactobacillus (p = 0.005), Stenotrophomonas (p < 0.001), Streptococcus (p < 0.001), Corynebacterium (p < 0.001), and Prevotella (p = 0.039) were abundant in bladder cancer urine samples. Midstream urine has a sensitivity of 83% for detecting dysbiotic bacteria in cancer tissue. Conclusions: Our research is the first microbiota study of bladder cancer done with naive patients who have never had an endourological intervention. Escherichia Shigella, Staphylococcus, Acinetobacter, Enhydrobacter, Delftia, Corynebacterium, and Pseudomonas were detected as dysbiotic bacteria in bladder cancer. The sensitivity of the midstream urine sample in detecting dysbiosis in tissue is 83%.

Vigor of bi-parametric MRI with MR segmentation unity in valuation of UB neoplasm mural invasion

BackgroundThe urinary bladder neoplasm is considered the second most frequent kind of neoplasm globally, after prostate cancer. Bladder neoplasm clinical management strategies and prognosis depend on the extent of loco-regional disease. MRI is increasingly being employed for preoperative, local staging of BC. The aim of this study was to expose the mastery of bi-parametric MRI (MRDWI and T2WI conjunction) with signal intensity-based MR segmentation in evaluation of UB neoplasm mural invasion if unaccompanied by contrast-enhanced MRI with special concern to Vesical Imaging Reporting and Data System (VIRADS) score taking the histopathological diagnosis as a reference.ResultsThis prospective study was conducted on 99 patients (with 100 UB lesions), who were referred from the urology department with bladder neoplasm suspicion. The mean age of the patients was 64.8 years. About 66.6% of the cases were smokers. The most common pathology of the UB neoplastic lesions was urothelial carcinoma. ADC in muscle-invasive bladder cancer (MIBC) was significantly lower than in non-invasive bladder cancer (NMIBC), with the minimum to maximum value of the ADC in all cases about 0.01–2.0. Also, we have done DWI at different b values, where b 1000 turned out to be the most accurate in detection of muscle invasion. Sensitivity, specificity, PPV, NPV and accuracy were calculated for T2WI only VIRADS, DWI only VIRADS, bi-parametric MR (T2WI and DWI)-based VIRADS, signal intensity-based MR segmentation VIRADS and for bi-parametric and MR segmentation conjunction-based VIRADS. The diagnostic performance for the combination of the bi-parametric MRI and signal intensity-based MR segmentations attained the highest values in sensitivity, specificity, PPV and NPV as well as accuracy.ConclusionsBi-parametric MRI (MRDWI with T2WI) and signal intensity MR segmentation conjunction have proven to be efficacious in accurately determining the UB neoplasm mural invasion allowing for the dispensability of CE-MRI in the event of contrast contraindications, unavailability or even its high cost.

Exploring a Desirable Quadr-mRNAs Panel for Non-invasive and Ultrasensitive Bladder Cancer Diagnosis: In-silico and Clinical Studies.

Most patients with non-muscle invasive bladder cancer (NMIBC) have a high direction for recurrence and disease progression, which remains a significant unresolved challenge in bladder cancer patients. Therefore, a constant search is necessary for identifying appropriate and reliable biomarkers for early diagnosis of NMIBC. The current study has aimed to search for valuable diagnostic biomarkers in the tissue and urine specimens of NMIBC patients. The changes of twelve candidate mRNAs in a screening phase (40 tissue samples of NMIBC patients and their corresponding 40 urine specimens) and a subsequent independent validation phase (40 urine specimens) were estimated using real-time polymerase chain reaction (RT-qPCR). The receiver operating characteristic (ROC) analysis was executed to determine the potential diagnostic values of mRNAs. The mRNA levels of seven candidate genes were markedly higher in tissue specimens relative to their neighboring tissues. Among them, four mRNAs, including ERBB2, CCND1, MKI67, and MAGEA6, were differentially expressed in urine samples of NMIBC patients relative to control subjects. Further, the expression of these four mRNAs was validated in the validation step. Combining these biomarkers showed better diagnostic performance than single biomarkers in the urine sample for non-invasive NMIBC detection. The combination of these mRNAs and cytology enhanced the sensitivity of cytology from 37% to 87%. Our findings suggested that a four-mRNA panel may be promising in the non-invasive diagnosis of NMIBC, which deserves further investigation.

The Role of Semaphorin 6D (Sema6D) in Non-Muscle-Invasive Bladder Cancer-A Preliminary Study on Human Plasma and Urine.

The incidence of bladder cancer worldwide in the last three decades has been increasing in both men and women. So far, there is no established non-invasive bladder cancer biomarker in daily clinical practice. Semaphorin 6D (sema6D) is a transmembrane protein that belongs to the class VI semaphorins. The aim of this study was to evaluate for the first time the potential role of sema6D in bladder cancer. The study group consisted of 40 patients with non-muscle-invasive bladder cancer (NMIBC) and the control group of 20 patients without malignancies. There was a statistically significantly higher urinary sema6D concentration in patients than controls (p < 0.05) but no significant difference in plasma 6D. There were no statistically significant differences in urinary or plasma concentration of sema6D between low- or high-grade cancer and according to the tumor stage in TNM classification. There was a statistically significant negative correlation between plasma sema6D and age of patients (R = -0.6; p = 0.019). Plasma sema6D does not seem to be useful in the clinical practice at this point. However, the urinary sema6D concentration could potentially serve as a marker of NMIBC used for diagnostic purposes, monitoring, and early relapse detection or the assessment of the treatment efficacy. Urinary sema6D is probably not associated with the grading or staging of NMIBC, so it cannot be used for the prediction of disease prognosis.

Insight into Fructose-to-Sucrose Ratio as the Potential Target of Urinalysis in Bladder Cancer.

Bladder cancer usually has been diagnosed in elderly patients as it stays asymptomatic until it presents. Current detection methods for bladder cancer cannot be considered as an adequate screening strategy due to their high invasiveness and low sensitivity. However, there remains uncertainty about targets with high sensitivity and specificity for non-invasive bladder cancer examination. Our study aims to investigate the actionable non-invasive screening biomarkers in bladder cancer. Here, we employed scRNA-seq to explore the crucial biological processes for bladder cancer development. We then utilized bidirectional Mendelian randomization (MR) analysis to explore the bidirectional causal relationship between ATP-associated metabolites in urine and bladder cancer. Lastly, we used a BBN-induced mouse model of bladder cancer to validate the crucial gene identified by scRNA-seq and MR analysis. We found that (1) the ATP metabolism process plays a critical role in bladder cancer development; (2) there is a bidirectional and negative causal relationship between fructose-to-sucrose ratio in urine and the risk of bladder cancer; and (3) the higher expression of TPI1, a critical gene in the fructose metabolism pathway, was validated in BBN-induced bladder tumors. Our results reveal that fructose-to-sucrose ratio can serve as a potential target of urinalysis in bladder cancer.

Objective Evaluation of Gaze Location Patterns Using Eye Tracking During Cystoscopy and Artificial Intelligence-Assisted Lesion Detection.

Background: The diagnostic accuracy of cystoscopy varies according to the knowledge and experience of the performing physician. In this study, we evaluated the difference in cystoscopic gaze location patterns between medical students and urologists and assessed the differences in their eye movements when simultaneously observing conventional cystoscopic images and images with lesions detected by artificial intelligence (AI). Methodology: Eye-tracking measurements were performed, and observation patterns of participants (24 medical students and 10 urologists) viewing images from routine cystoscopic videos were analyzed. The cystoscopic video was captured preoperatively in a case of initial-onset noninvasive bladder cancer with three low-lying papillary tumors in the posterior, anterior, and neck areas (urothelial carcinoma, high grade, and pTa). The viewpoint coordinates and stop times during observation were obtained using a noncontact type of gaze tracking and gaze measurement system for screen-based gaze tracking. In addition, observation patterns of medical students and urologists during parallel observation of conventional cystoscopic videos and AI-assisted lesion detection videos were compared. Results: Compared with medical students, urologists exhibited a significantly higher degree of stationary gaze entropy when viewing cystoscopic images (p < 0.05), suggesting that urologists with expertise in identifying lesions efficiently observed a broader range of bladder mucosal surfaces on the screen, presumably with the conscious intent of identifying pathologic changes. When the participants observed conventional and AI-assisted lesion detection images side by side, contrary to urologists, medical students showed a higher proportion of attention directed toward AI-detected lesion images. Conclusion: Eye-tracking measurements during cystoscopic image assessment revealed that experienced specialists efficiently observed a wide range of video screens during cystoscopy. In addition, this study revealed how lesion images detected by AI are viewed. Observation patterns of observers' gaze may have implications for assessing and improving proficiency and serving educational purposes. To the best of our knowledge, this is the first study to utilize eye tracking in cystoscopy. University of Tsukuba Hospital, clinical research reference number R02-122.

Abstract LB437: Development of a bladder cancer-on-a-chip assay to assess the invasive potential of bladder cancer cells

Abstract Goals: The primary goal of this study was to develop a 3D functional bioassay to assess the relative invasiveness of bladder cancer cells. This model will support the testing of migrastatic drugs and investigation of the molecular mechanisms which allow for bladder cancer cell invasion to occur. Methods: Our model uses an organ-on-a-chip platform to assess bladder cancer cell invasiveness. This platform supports 3D culture of cells as well as pharmacokinetic studies. Our chips were designed to have a center channel to hold cells supported in Matrigel®, 2 adjacent side channels filled with Matrigel® only, and 2 outer media channels. Total cell and Matrigel® side channel length is 3 cm, channel height is 150 µm, and each channel width is 1 mm. Four bladder cancer cell lines (MB49, T24, J82, RT4) and cells isolated from a mouse bladder were used to develop our model. Cells were mixed with 1:1 media and Matrigel® and a total of 5,000 cells/µl introduced into the center channel. The 2 adjacent side channels were then filled with Matrigel®. The upper media channel was filled with 10% FBS cell culture media while the lower media channel was filled with 1% charcoal stripped cell culture media to thereby create a nutrient and growth factor gradient. Invasion of cells into the Matrigel®-containing side channels was assessed every 24 hours. Using the ImageXpress machine (Molecular Dynamics), we measured the furthest distance travelled by the cells. The chips were then fixed in 4% paraformaldehyde and stained using FITC-phalloidin and DAPI. Results: J82 and T24 cell lines, derived from patients with muscle invasive bladder cancer, and the MB49 cell line, a highly invasive mouse bladder cancer cell line, started invading into the Matrigel® side channels within 24 hours. T24, J82, and MB49 cells moved a maximum of 110, 87, and 170 µm by 48 hours, respectively. In contrast, RT4 cells, derived from a patient with non-invasive bladder cancer, took 48 hours to invade the Matrigel® channels and only invaded a maximum of 3 µm. Cells isolated from a mouse bladder did not invade into the Matrigel® side channels but survived within the chips for over 20 days. Staining of chips with DAPI and FITC-phalloidin allowed for clear visualization of cells and the actin cytoskeleton and demonstrated that immunostaining of cells within chips is possible. Conclusions: Our chip model allows for studying the invasiveness of bladder cancer cells and immunostaining experiments which can be used to assess the molecular mechanisms which drive invasion. It can also support the growth and assessment of primary cells. This model has the potential to improve bladder cancer patient treatments and outcomes by providing a physiologically relevant system to test new and existing drug combinations and to better understand the molecular mechanisms which drive bladder cancer cell invasion. Citation Format: Desiree Ewell, Nita Vue, Sakib Moinuddin, Tanoy Sarkar, Fakhrul Ahsan, Ruth L. Vinall. Development of a bladder cancer-on-a-chip assay to assess the invasive potential of bladder cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB437.

Diabetes and the risk of bladder cancer subtypes in men and women: results from the Netherlands Cohort Study

Meta-analyses have shown modest positive associations between diabetes mellitus (DM) and bladder cancer risk, but results are heterogeneous. This might be due to lack of distinction between bladder cancer subtypes, between sexes, and possibly between Type 2 and Type 1 DM (T2DM and T1DM). The relationship of T2DM (and secondarily T1DM) characteristics with risk of bladder cancer subtypes (invasive versus noninvasive) was investigated in the Netherlands Cohort Study. In 1986, 120,852 men and women aged 55–69 years provided information on DM and lifestyle data. After 20.3 years of follow-up, multivariable case-cohort analyses were based on 1020 invasive and 1088 noninvasive bladder cancer cases, and 4267 subcohort members with complete data on DM and confounders. While T2DM was not associated with noninvasive bladder cancer, it was statistically significantly associated with invasive bladder cancer risk: the multivariable-adjusted was HR = 1.57 (95% CI 1.04–2.37), comparing participants with T2DM versus without DM. The association was only significant in women, and women showed a stronger association [HR = 2.19 (95% CI 1.10–4.34)] between T2DM and invasive bladder cancer than men [HR = 1.42 (95% CI 0.88–2.30)]; interaction by sex was nonsignificant. Associations were stronger positive in those whose age at diagnosis of T2DM was 55+ years, and in those diagnosed with T2DM less than five years before baseline. T2DM participants using antidiabetic medication had higher invasive bladder cancer risk than those without DM. Exploratory age-sex-adjusted analyses suggested a positive association between T1DM and invasive bladder cancer, but this was based on few cases. These findings suggest that T2DM and possibly T1DM are positively associated with invasive bladder cancer risk.

Targeting cancer-derived extracellular vesicles by combining CD147 inhibition with tissue factor pathway inhibitor for the management of urothelial cancer cells

BackgroundExtracellular vesicles (EVs), including microvesicles, hold promise for the management of bladder urothelial carcinoma (BLCA), particularly because of their utility in identifying therapeutic targets and their diagnostic potential using easily accessible urine samples. Among the transmembrane glycoproteins highly enriched in cancer-derived EVs, tissue factor (TF) and CD147 have been implicated in promoting tumor progression. In this in vitro study, we explored a novel approach to impede cancer cell migration and metastasis by simultaneously targeting these molecules on urothelial cancer-derived EVs.MethodsCell culture supernatants from invasive and non-invasive bladder cancer cell lines and urine samples from patients with BLCA were collected. Large, microvesicle-like EVs were isolated using sequential centrifugation and characterized by electron microscopy, nanoparticle tracking analysis, and flow cytometry. The impact of urinary or cell supernatant-derived EVs on cellular phenotypes was evaluated using cell-based assays following combined treatment with a specific CD147 inhibitor alone or in combination with a tissue factor pathway inhibitor (TFPI), an endogenous anticoagulant protein that can be released by low-molecular-weight heparins.ResultsWe observed that EVs obtained from the urine samples of patients with muscle-invasive BLCA and from the aggressive bladder cancer cell line J82 exhibited higher TF activity and CD147 expression levels than did their non-invasive counterparts. The shedding of GFP-tagged CD147 into isolated vesicles demonstrated that the vesicles originated from plasma cell membranes. EVs originating from invasive cancer cells were found to trigger migration, secretion of matrix metalloproteinases (MMPs), and invasion. The same induction of MMP activity was replicated using EVs obtained from urine samples of patients with invasive BLCA. EVs derived from cancer cell clones overexpressing TF and CD147 were produced in higher quantities and exhibited a higher invasive potential than those from control cancer cells. TFPI interfered with the effect when used in conjunction with the CD147 inhibitor, further suppressing homotypic EV-induced migration, MMP production, and invasion.ConclusionsOur findings suggest that combining a CD147 inhibitor with low molecular weight heparins to induce TFPI release may be a promising therapeutic approach for urothelial cancer management. This combination can potentially suppress the tumor-promoting actions of cancer-derived microvesicle-like EVs, including collective matrix invasion.Graphical

The Value of Negative Urinary Dipstick Tests for Haematuria in Patients Undergoing Surveillance for Low-grade Ta Urothelial Cancer: A Two-stage Prospective Clinical Study in 524 Patients

Background and objectiveThe risk of first recurrence beyond 5 yr for patients with low-grade (LG) Ta non–muscle-invasive bladder cancer (NMIBC) is low enough to consider discontinuing cystoscopic surveillance at that point. However, a positive urinary dipstick test for haematuria (UDH) during and beyond the period of cystoscopic surveillance can disrupt plans to cease surveillance because the association between UDH positivity and recurrence in LG Ta NMIBC is unknown. In a two-stage study, we evaluated this association and explored the role of UDH negativity in predicting the absence of recurrence. MethodsBecause of previously demonstrated changes in recurrence patterns over time, two prospective cohorts were assessed: an “exploratory” cohort (January 2007–March 2008) and a “validation” cohort (November 2017–August 2018). UDH was performed before flexible cystoscopy. Patient, operative, and surveillance data have been recorded prospectively using standard pro forma sheets since 1978 in our institution. Only patients with primary LG Ta pTa NMIBC were included for analysis. Key findings and limitationsWe assessed 231 patients in the exploratory group and 293 in the validation group. The proportion of smokers (67% vs 70%; p = 0.5) and mean follow-up (72.2 vs 79.9 mo; p = 0.2) were similar between the groups. The recurrence rate was higher in the exploratory group (19% vs 11%; p = 0.009), as was the UDH positivity rate (37% vs 11%; p < 0.001). The specificity and negative predictive value were 64% and 83% in the exploratory group, and 90% and 90%, respectively, in the validation group. These values increased further for the subgroup with solitary primary tumours the subgroup without recurrence for 3 yr. Conclusions and clinical implicationsUDH negativity has a high probability of being associated with the absence of recurrence in small LG Ta NMIBC and could be an inexpensive adjunct during surveillance. Ongoing validation, which started in 2019, is being performed in a now-nationalised Scottish protocol in which UDH replaces cystoscopy in years 2 and 4 for patients in the low-risk group. Patient summaryWe investigated the accuracy of a dipstick test for blood in the urine for patients undergoing surveillance for low-grade noninvasive bladder cancer. We found that a negative dipstick test result was highly associated with the absence of tumour recurrence, particularly for patients with the lowest risk. These findings have been introduced into a national protocol designed to reduce the frequency of telescopic inspection of the bladder during surveillance to reduce the burden for patients.

Mathematical model of MMC chemotherapy for non-invasive bladder cancer treatment.

Mitomycin-C (MMC) chemotherapy is a well-established anti-cancer treatment for non-muscle-invasive bladder cancer (NMIBC). However, despite comprehensive biological research, the complete mechanism of action and an ideal regimen of MMC have not been elucidated. In this study, we present a theoretical investigation of NMIBC growth and its treatment by continuous administration of MMC chemotherapy. Using temporal ordinary differential equations (ODEs) to describe cell populations and drug molecules, we formulated the first mathematical model of tumor-immune interactions in the treatment of MMC for NMIBC, based on biological sources. Several hypothetical scenarios for NMIBC under the assumption that tumor size correlates with cell count are presented, depicting the evolution of tumors classified as small, medium, and large. These scenarios align qualitatively with clinical observations of lower recurrence rates for tumor size ≤ 30[mm] with MMC treatment, demonstrating that cure appears up to a theoretical x[mm] tumor size threshold, given specific parameters within a feasible biological range. The unique use of mole units allows to introduce a new method for theoretical pre-treatment assessments by determining MMC drug doses required for a cure. In this way, our approach provides initial steps toward personalized MMC chemotherapy for NMIBC patients, offering the possibility of new insights and potentially holding the key to unlocking some of its mysteries.

Prognostic value of a panel of immunohistochemical markers for determining the risk of recurrence and progression of non-invasive bladder cancer

Background. There is a huge number of algorithms for the diagnosis and prognosis of the clinical course of non-invasive bladder cancer (NIBC). They are based on both clinical and morphological features and data from immunohistochemical (IHC) studies in which a large number of markers, including p53, Ki-67, VEGF, E-cadherin, MMP, bcl, etc., were used. Of scientific and practical interest is a comprehensive assessment of the prognostic value of a number of the main IHC markers used for the diagnosis and prognosis of bladder cancer, the impact of epithelial-mesenchymal transition (EMT) processes and the immune response on the risk of recurrence and tumor progression. Purpose. To study the prognostic value of the panel of IHC markers for determining the risk of recurrence and progression of NIBC. Materials and Methods. We studied surgically removed stage T1 malignant bladder tumors that spread to the submucosal tissue without invasion into the muscle tissue in 42 patients. Recurrence was understood as tumor development within 5 years after surgical treatment. The term «progression» meant the growth of the tumor in the muscle tissue, in other words, transformation of the tumor from non-muscle-invasive to muscle-invasive, which is of great prognostic significance for this type of cancer. IHC study was performed using primary ready-to-use monoclonal antibodies produced by DAKO (Denmark) for p53, p63, Ki67, E-cadherin, N-cadherin, CK7, CK20, Vimentin (VimentinV9), MMP-9, TGF- β1, VEGF, CD34+, CD3+, CD4+, CD8+, CD20+, CD68+, collagen IV. Epithelial markers (E-cadherin, cytokeratins 20 (CK20) and 7 (CK7)) and mesenchymal markers (N-cadherin and vimentin) were used to study EMT processes. Statistical processing of the research results was performed using the Statistica 6.0 package. Results and Discussion. It was found that the following markers may have prognostic value for the recurrence of NIBC: p53, CD3+, CD8+, CD68+; for recurrence with progression: Ki-67 and MMP-9. As differentiation of NIBC decreased, we observed increased expression of vimentin, Ki-67, MMP-9, VEGF, CD3+, CD8+, CD68+, p53, CD 34 and decreased expression of E-cadherin, CK20, collagen IV, p63. We established a relationship between the clinical behavior of NIBC and EMT processes. Thus, stage 1 EMT was characteristic of cancer with recurrence but without progression (p &lt; 0.05), and stage 2 EMT was characteristic of NIBC with recurrence and progression (p &lt; 0.05). The association of EMT with immune cell infiltration of NIBC was proven. The emergence of EMT in NIBC is associated with an increase in the intensity of infiltration by CD68+ macrophages (p &lt; 0.01), CD3+-T-lymphocytes (p &lt; 0.05), including both CD8+ (p &lt; 0.01) and CD4+ (p &lt; 0.05) cells. Conclusions. Non-progressive cancer with recurrence is characterized by stage 1 EMT of the tumor cells (p &lt; 0.05), and progressive NIBC with recurrence is characterized by stage 2 EMT (p &lt; 0.05). The association of EMT with immune cell infiltration of NIBC was proven. The emergence of EMT in NIBC is associated with an increase in the intensity of infiltration by CD68+ macrophages (p &lt; 0.01), CD3+ T-lymphocytes (p &lt; 0.05), including both CD8+ (p &lt; 0.01) and CD4+ (p &lt; 0.05) cells.

Proteomics Profiling of Bladder Cancer Tissues from Early to Advanced Stages Reveals NNMT and GALK1 as Biomarkers for Early Detection and Prognosis of BCa.

The high recurrence rate and invasive diagnostic and monitoring methods in bladder cancer (BCa) clinical management require the development of new non-invasive molecular tools for early detection, particularly for low-grade and low-stage BCa as well as for risk stratification. By using an in-solution digestion method and label-free data-independent LC-MS/MS coupled with ion mobility, we profiled the BCa tissues from initiation to advanced stages and confidently identified and quantified 1619 proteins (≥2 peptides). A statistically significant difference in abundance (Anova ≤ 0.05) showed 494 proteins. Significant correlation with stage with steady up or down with BCa stages showed 15 proteins. Testing of NNMT, GALK1, and HTRA1 in urine samples showed excellent diagnostic potential for NNMT and GALK1 with AUC of 1.000 (95% CI: 1.000-1.000; p < 0.0001) and 0.801 (95% CI: 0.655-0.947; p < 0.0001), respectively. NNMT and GALK1 also showed very good potential in discriminating non-invasive low-grade from invasive high-grade BCa with AUC of 0.763 (95% CI: 0.606-0.921; p = 0.001) and 0.801 (95% CI: 0.653-0.950; p < 0.0001), respectively. The combination of NNMT and GALK1 increased prognostic accuracy (AUC = 0.813). Our results broaden the range of potential novel candidates for non-invasive BCa diagnosis and prognosis.

Single-Dose Post-Office Fulguration Mitomycin C Instillation Appears to Improve Recurrence-Free Survival in Patients With Low-Grade Noninvasive Bladder Cancer.

Perioperative instillation of mitomycin-C (MMC) has shown effectiveness in reducing the recurrence of low-grade non-muscle invasive bladder cancer (NMIBC). Data is lacking about the impact of single-dose MMC following office fulguration of low-grade urothelial carcinoma. We compared the outcomes of small-volume low-grade recurrent NMIBC in patients treated with office-fulguration - with and without an immediate single-dose instillation of MMC. A retrospective analysis of medical records of patients with recurrent small-volume (≤1 cm) low-grade papillary urothelial cancer who underwent fulguration in a single institution between January 2017 and April 2021 either with or without instillation of post-fulguration MMC (40mg/50 mL). The primary outcome was recurrence-free survival (RFS). Of 108 patients (27% women) who underwent fulguration, 41% received intravesical MMC. The treatment and control groups had similar sex ratio, mean age, mass size, tumor multifocality and or tumor grade. Median RFS was 20 months (95% CI 4-36) in the MMC group and 9 months (95% CI 5-13) in the control group (P=.038). Multivariate Cox regression analysis showed that MMC instillation was associated with longer RFS (OR=0.552, 95% CI 0.320-0.955, P=.034) and multifocality was associated with shorter RFS (OR=1.866, 1.078-3.229, P=.026). A higher rate of grade 1-2 adverse events was observed in the MMC group (18.2%) vs. the control (6.8%, P=.048). No complications grade 3 or higher were observed. A single dose of MMC instilled after office fulguration is associated with longer RFS compared to patients who did not receive MMC after the procedure, with no associated high-grade complications.

Cluster of differentiation 147 (CD147) as potential membrane protein biomarker for bladder cancer cells

Studies reveal that alterations in membrane protein (MP) patterns are associated with underlying drug resistance to chemotherapy. Therefore, the tryptic-digested MPs from the bladder cancer cell line were subjected to global proteomics using LC-MS/MS to identify the highly expressed potential MPs in bladder cancer cells. Our findings revealed the identification of MP biomarkers, CD147, and caveolin-1. Immunocytochemistry analysis confirmed the presence of CD147 on the cell membrane, while caveolin-1 showed positive signals without apparent staining on the membrane, suggesting its existence in multiple locations. Western blot analysis confirmed the higher expression of CD147 in non-invasive (RT 112) and metastatic (UM-UC-13) bladder cancer cells compared to invasive bladder cancer cells (5637 and J82), suggesting its potential as an MP biomarker for both of the former subtypes. The identified MPs could be used as drug therapy targets aimed at improving drug sensitivity and enhancing treatment outcomes in bladder cancer patients. SignificanceIdentification of the membrane proteins associated with bladder cancer recurrence is crucial to understanding the mechanisms underlying the drug resistance to chemotherapy.

Enzyme-free nucleic acid and luminescent material cascade amplifications enable urinary dual miRNAs assay for non-invasive bladder cancer diagnosis

We presented a facile biosensing strategy for the detection of dual microRNAs (miRNA-21 and miRNA-126) in urine samples from bladder cancer (BC) patients. The fluorescent homogeneous detection of miRNAs was achieved by elaborate parallel catalytic hairpin assembly (CHA) integrated with material cascade amplifications. The continuous release of G-quadruplex (G4) sequences and Hg2+ was attributed to the specific recognition between the miRNAs and the corresponding hairpin oligonucleotides. Assisted by the streptavidin-(T-Hg2+-T-biotin)4 structures, more Hg2+ emission was obtained, and the spatial site block effect was used to order its distribution, significantly improving the selective recognition performance of cadmium telluride quantum dots (CdTe QDs) (1.4–2.9 times). Furthermore, the peroxidase-like activity of G4-hemin was used to catalyze the oxidation of dopamine (DA) to fluorescence-enhanced polydopamine (PDA). The generated PDA signal and the Hg2+-quenched CdTe QDs signal completed the synchronous detection to achieve a high-sensitivity analysis of the dual miRNAs. The limit of detection (LOD) of both miRNAs was as low as the fM level. Eventually, 56 clinical urine samples were quantified. The results showed satisfying specificity (100%) and sensitivity (92%) with a strong clinical agreement, including computed tomography (CT) and pathology findings, indicating a rapid non-invasive solution for early diagnosis of BC.

Vesical Imaging-Reporting and Data System (VI-RADS): Inter-Observer Agreement and Impact of Reader's Experience

Background The Vesical Imaging Reporting and Data System (VI-RADS) is based on multiparametric MRI (mpMRI), to provide accurate information to differentiate between muscleinvasive bladder cancer (MIBC) and non-muscle –invasive bladder cancer (NMIBC). Aim of the Work To evaluate inter-observer agreement according to observer experience and diagnostic performance of the new scoring system based on magnetic resonance imaging called Vesical Imaging-Reporting and Data System (VI-RADS) to differentiate between invasive and non-invasive bladder cancer. Patients and Methods This study included 38 patients who were diagnosed with Transitional Cell Carcinoma (TCC) who underwent multiparametric Magnetic Resonance Imaging (mpMRI) at the department of Radiodiagnosis in Ain-Shams University from April 2021 till October 2021. Three radiologists with different levels of experience reviewed and scored all MRI examinations according to VI-RADS, compared them with the histopathological findings and inter-observer agreement was assessed. Results This study included 38 patients; the patients’ age ranged from 47 to 83 years old with mean age 62.26 (+8.18 SD). The total number of lesions in all cases were 43; 12 of them (27.9 %) were non-muscle-invasive urothelial carcinoma and 31 (72.1%) were muscle invasive. Using a VI-RADS score of 4 or greater as the cutoff value, the sensitivity, specificity, accuracy, PPV, and NPV for reader 1 were 96.8%, 83.3 %, 93%, 93.8%, and 90.9%, and for reader 2 were 93.5 %, 91.7%, 93%, 96.7% and 84.6%, and finally for reader 3 were 96.8%, 83.3%, 93 %, 93.8% and 90.9%. The agreement was excellent between each reader with the pathology and the readers with each other (Kappa over 0.75). Conclusion The Vesical Imaging-Reporting and Data System (VI-RADS) is an effective comprehensive scoring system, with satisfactory sensitivity, specificity, and diagnostic value for detecting detrusor muscle invasion. We can recommend VI-RADS scoring for use in the prediction of detrusor muscle invasion in patients with bladder cancer, but further multicenter validation studies containing multiple magnetic field strength devices also must be conducted.