Urothelial Tumors Research Articles

VISIOCYT1 clinical trial: Artificial intelligence for the diagnosis of bladder urothelial lesions.

e16558 Background: Voided urinary cytology is a non-invasive test, but it suffers from a lack of sensitivity (Se), particularly in low-grade urothelial lesions, and it remains pathologist-dependent. In order to improve the performance of urinary cytopathology, the VitaDX company (France) developed a medical device which uses whole-slide digitalization and artificial intelligence algorithms to identify tumor cells. The device allows complete analysis of morphological characters of voided urothelial cells, such as the shape, size and color of the nuclei. Methods: VISIOCYT1 is a prospective, multicenter clinical trial involving 319 patients divided into two groups: 1) patients with non-muscle invasive bladder tumors (NMIBC) confirmed by histology, and 2) control patients with negative urinary cytology and cystoscopy. The primary objective of the study was to evaluate the Se of the VisioCyt test. Specificity (Sp) and comparison of Se and Sp of conventional urinary cytology vs. the VisioCyt test were also calculated. Results: A total of 319 patients were included (170 in the NIMBC group vs. 149 controls). Overall Se was 80.9% and 45.9% for VisioCyt test and conventional urinary cytology, respectively (p = .002). Negative cytology being an inclusion criterion, a 100% Sp was attributed to the control group. It was calculated at 61.8% for the VisioCyt test. Concerning grade, the VisioCyt test allowed higher Se to be obtained in the low grade tumor category (66.7% vs. 26.1% for conventional urinary cytology, Mc Nemar test, p < .0001). A higher Se was also observed in high grade tumors: 93.7% vs. 62.8% (p < .0001). Conclusions: The VisioCyt test greatly improves the performance of urinary cytopathology in the diagnosis of urothelial bladder tumors, particularly in low grade, pTa lesions. Its implementation could dramatically reduce the need for cystoscopy in patients followed after conservative treatment of bladder cancer. Clinical trial information: NCT02966691. [Table: see text]

Phase 1a/b open-label study of IK-175, an oral AHR inhibitor, alone and in combination with nivolumab in patients with locally advanced or metastatic solid tumors and urothelial carcinoma.

TPS3169 Background: Aryl Hydrocarbon Receptor (AHR) is a ligand-activated transcription factor that regulates the activity of multiple innate and adaptive immune cells. Kynurenine, generated from tryptophan by IDO1 and TDO2, is a ligand that binds AHR and leads to a net immunosuppressive tumor microenvironment, making AHR an attractive therapeutic target in multiple cancer types. IK-175 is a selective, small molecule inhibitor of AHR being developed as an oral agent. In preclinical mouse tumor models, IK-175 demonstrates antitumor activity as a single agent or in combination with checkpoint inhibitors. AHR immunohistochemistry (IHC) tumor microarray analysis across 15 different tumor types revealed that bladder cancer has the highest level of AHR protein expression and nuclear localization indicative of ligand-bound and active AHR signaling. Therefore, nuclear AHR in urothelial carcinoma tumors is being investigated for potential predictive clinical benefit with IK-175. Methods: This is a first-in-human, phase 1a/b, open-label, multicenter, dose-escalation and expansion study of IK-175 as a single agent and in combination with nivolumab. The primary objectives are to determine the maximum tolerated dose (MTD) and/or maximum administered dose (MAD), identify the recommended phase 2 dose (RP2D), and evaluate the safety and tolerability of IK-175 alone and in combination with nivolumab. Secondary objectives are to evaluate the pharmacokinetics (PK) of IK-175, evaluate pharmacodynamic (PD) immune effects, and assess preliminary antitumor activity. Key exploratory objectives are to evaluate the PD effects on peripheral immune cells and target gene expression, to assess candidate baseline biomarkers, and correlative analyses of tumor AHR nuclear localization with clinical response. The study is exploring tumor AHR nuclear localization by IHC as a predictive biomarker in patients with urothelial carcinoma. A minimum of 10 patients with a positive AHR nuclear localization test (cutoff for positive AHR is 65% tumor cells positive for 2+/3+ nuclear AHR by a validated IHC assay) will be enrolled in the combination arm. IK-175 is administered daily in 21 or 28 day-cycles as a single agent, and in combination with nivolumab (480 mg q4w on Day 1 of every cycle), in adult patients with advanced solid tumors (dose escalation) and urothelial carcinoma (dose expansion). Key eligibility criteria include patients with histologically confirmed solid tumors (including urothelial carcinoma) who have locally recurrent or metastatic disease that have progressed on or following all standard of care therapies deemed appropriate by the treating physician including prior checkpoint inhibitors. Estimated enrollment is 93 patients; the study began in January 2020 and is ongoing. Clinical trial information: NCT04200963.

Genomic and immunologic profiles of concurrent RB1 and CDKN1A/p21(WAF1) truncating mutations (RW+) in bladder cancer.

4571 Background: p53 target and cell cycle inhibitor CDKN1A/p21(WAF1) was initially not found to be mutated in cancer. TCGA analysis identified CDKN1A mutations are present but rare with frequencies of < 1%, but enrich in bladder cancer (̃8%). Truncating WAF1 mutations are associated with sensitivity to cisplatin and are associated with truncating Rb mutations in bladder cancer (RW+). We hypothesized RW+ bladder cancers may represent a unique subgroup with sensitivity to therapeutics. Methods: A total of 1104 urothelial tumors underwent molecular profiling at Caris Life Sciences (Phoenix, AZ) utilizing NGS of DNA (592 Gene Panel, NextSeq, or WES, NovaSeq) and RNA (NovaSeq, WTS). Wilcoxon, Fisher’s exact were used for statistical significance (p value without and q value with multi comparison correction). Immune cell fraction (QuanTIseq) and pathway analysis (ssGSEA) were assessed by mRNA analysis. Immune epitope prediction was performed using the NetMHCpan v4.0 method in the Immune Epitope Database. Results: Concurrent truncating mutation (frameshift, nonsense) for RB1 and WAF1 were detected in 47 tumors (RW+, 4.25%) and tumors with wild-type status for both RB1 and WAF1 genes were classified as RW- group (54.08%). Tumors harboring only one RB1 or WAF1 mutation were excluded for further analysis. When compared to RW- group, RW+ tumors showed lower mutation rate of TP53 (54.5% vs 80.9%, q < 0.05), ARID1A (23.5% vs 38.3%, p < 0.05), and PIK3CA (18.4% vs 31.9%, p < 0.05). Interestingly, RW+ was mutually exclusive with FGFR3 mutation (18.0% vs 0%, p < 0.05). We further evaluated RNA expression of DNA repair and checkpoint arrest pathways. Notably, E2F pathway (Normalized Enrichment Scores, NES: 0.89 vs 0.86, q < 0.01) and DNA G2M checkpoint (NES: 0.89 vs 0.86, q < 0.01) were found to be the most enriched in RW+ with respect to RW- group. In addition, mRNA levels of FANCC/A, CHEK1, WEE1, CDC25A/C, PALB2 and BRCA1/2 were found to be overexpressed in RW+ group (q < 0.05). RW+ tumors also displayed a distinct immunological profile: They were associated with higher PD-L1 status (63.8% vs 37.3%, q < 0.01), higher median TMB (11 mut/Mb vs 8 mut/Mb, q < 0.01) and with less frequent loss of heterozygosity for HLA-DPA1 (51.1% vs 66.7%, p < 0.05), with more high-binding-affinity neoantigen load (4.78 vs 3.89, p < 0.05) to MHC proteins, consistent with the significantly more myeloid dendritic cells in in RW+ group (0.3 vs 0.04, q < 0.001). Conclusions: Concurrent truncating mutation in RB1 and WAF1 (RW+) bladder carcinomas have fewer p53, ARID1A, and PIK3CA mutations but are enriched for E2F targets, G2/M checkpoint genes, FANCC/A, CHEK1, WEE1, CDC25A/C, PALB2 and BRCA1/2 and have a distinct immunological profile. The findings suggest therapeutic strategies for RW+ bladder cancers including Chk1/Wee1, PARP inhibitors, -/+ immunotherapy that may impact on clinical outcomes.

Novel use of ctDNA to identify muscle-invasive and non-organ-confined upper tract urothelial carcinoma.

4587 Background: Upper tract urothelial carcinoma (UTUC) is an aggressive cancer for which use of neoadjuvant chemotherapy (NAC) is limited by suboptimal clinical staging prior to nephroureterectomy. Detection of circulating tumor DNA (ctDNA) is associated with locally advanced and metastatic urothelial carcinoma of the bladder and may help identify UTUC patients who would benefit from NAC. Here we examine the feasibility and utility of plasma ctDNA in the diagnosis of non-organ confined high-risk UTUC. Methods: Patients with high-grade cTa-T2 UTUC without radiographic evidence of metastatic disease undergoing up-front radical nephroureterectomy (RNU) were prospectively accrued for pre- and post-operative plasma collection. Blood was collected preoperatively on the day of surgery, and plasma and buffy coat were processed for extraction of cell-free DNA and genomic DNA, respectively. FFPE tumor samples from RNU were used for tissue genomic DNA extraction. Targeted next-generation sequencing (NGS) was used for variant profiling. ctDNA positivity was defined as the presence of plasma cell-free DNA variants concordant with tissue-based variants. Results: NGS analyses of matched FFPE and plasma samples were successfully performed for all 19 accrued UTUC patients. Alterations in the TERT promoter (74%), TP53 (58%), FGFR3 (53%), myc amplification (53%), and ATM (42%) were demonstrated in urothelial tumor tissue. Matched plasma ctDNA showed prevalent alterations in the TERT promoter (42%), TP53 (42%), PIK3CA (37%), ATM (32%) and CD274 (26%). Nine patients (47%) had detectable plasma ctDNA mutations concordant with tumor-specific variants using the targeted NGS panel. All patients with detectable preoperative ctDNA had advanced staging (≥pT2 or ≥pN1) and lymphovascular invasion on final pathology, resulting in a 90% sensitivity. The panel was 100% specific with no patients with pTis, pTa, pT1 and pN0 having detectable concordant ctDNA mutations. Concordant plasma ctDNA was detected in four of nine patients postoperatively. Two of three (67%) who developed metastatic disease had detectable ctDNA while neither of the two who developed non-muscle-invasive bladder recurrences did. Conclusions: Prospective ctDNA analysis using a targeted NGS panel can be used to predict muscle-invasive and non-organ-confined UTUC preoperatively. Detectable postoperative ctDNA may indicate residual disease and predate clinical recurrence.

Overexpression of long intergenic noncoding RNAs in bladder cancer: A new insight to cancer diagnosis

There is increasing evidence that show long noncoding RNAs including long intergenic noncoding RNAs (lincRNA) play a pivotal regulatory role in the biological processes. Differential expression of lincRNAs can be utilized for cancer diagnosis, prognosis, and targeted therapy. Little is known about their expressions in urothelial tumors. Concerning the potential role of lincRNAs in cancer development, we aimed to investigate the expression levels of LINC00958 and DNM3OS in bladder cancer. Fifty tumor and 50 adjacent non-tumor tissue samples along with their clinicopathological parameters were obtained from bladder cancer patients. Expressions of LINC00958 and DNM3OS were analyzed by Real-time PCR. ROC curve analysis was used to evaluate the diagnostic power of LINC0095 and DNM3OS for BC. Expression level of LINC00958 was considerably increased in cancerous tissues (P < 0.001) and in correlation with cigarette smoking (P = 0.043). DNM3OS expression was higher in the tumor tissues than normal tissues (P < 0.001) and showed a significant association with age (P = 0.038). By using the ROC curve, the diagnostic power of LINC00958 and DNM3OS transcript levels in bladder cancer were estimated to be 87% and 75%, respectively. Our findings offer some important intuitions into the oncogenic role of LINC00958 and DNM3OS in bladder cancer and suggest that they can be candidate biomarkers and may provide new approaches for the diagnosis and therapy if being validated in a larger sample size of clinical samples as well as functional studies.

Urinary bladder tumour in a 10 years old child: A case report on urothelial papilloma

The suspicion and diagnosis of urothelial neoplasm are considered a rare commodity in the pediatric age group presenting with hematuria. A 10 years old patient presented to out-patient department with lower pain abdomen and hematuria for 2 months. Routine examination and ultrasonography, followed by urethrocystoscopy, revealed a solitary intravesical lesion in close approximation to the right ureteric orifice. Patient underwent transurethral resection of the bladder mass with histopathology confirming urothelial papilloma. This benign tumour is extremely rare in childhood with much to decipher of its etiology and further management.

Large-scale human tissue analysis identifies Uroplakin 1b as a putative diagnostic marker in surgical pathology

Uroplakin 1B (Upk1b) stabilizes epithelial cells lining the bladder lumen to prevent rupturing during bladder distension. Little is known about Upk1b expression in other normal and malignant tissues. To comprehensively evaluate the potential diagnostic and prognostic utility of Upk1b expression analysis, a tissue microarray containing 14,061 samples from 127 different tumor types and subtypes and 608 samples of 76 different normal tissue types was analyzed by immunohistochemistry. Upk1b immunostaining was found in 61 (48%) different tumor types including 50 (39%) with at least one moderately positive and 39 tumor types (31%) with at least one strongly positive tumor. Highest positivity rates were found in urothelial neoplasms (58-95%), Brenner tumors of the ovary (92%), epithelioid mesothelioma (87%), serous carcinoma of the ovary (58%) and the endometrium (53%) as well as in squamous cell carcinoma of the head and neck (18-37%), lung (39%), and esophagus (26%). In urothelial carcinoma, low Upk1b expression was linked to high grade and invasive tumor growth (P<.0001 each) and nodal metastasis (P=.0006). Our data suggest diagnostic applications of Upk1b immunohistochemistry in panels for the distinction of malignant mesothelioma from adenocarcinoma of the lung, urothelial carcinoma from prostatic adenocarcinoma in the bladder, or pancreaticobiliary and gastroesophageal from colorectal adenocarcinoma.

PD12-11 CASTRATION IMPACT ON PATTERN RECOGNITION RECEPTORS (PRRS) BOOSTS TOLL-LIKE RECEPTOR-4 EXPRESSION IN AN IMMUNOCOMPETENT BLADDER CANCER MALE MODEL

You have accessJournal of UrologyCME1 May 2022PD12-11 CASTRATION IMPACT ON PATTERN RECOGNITION RECEPTORS (PRRS) BOOSTS TOLL-LIKE RECEPTOR-4 EXPRESSION IN AN IMMUNOCOMPETENT BLADDER CANCER MALE MODEL Leonardo Reis Leonardo ReisLeonardo Reis More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002538.11AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Among diverse Pattern Recognition Receptors (PRRs), the Toll-Like Receptor-4 (TLR-4) is the key urothelial trigger for innate immune response and plays a central role as Bacillus Calmette Guerin (BCG) main target. Also, its gene polymorphism and expression are associated to bladder cancer (BCa) prognosis. Androgen activation promotes immunotolerance, playing an immunoregulatory role, though by unknown mechanisms. METHODS: We described an efficient male immunocompetent BCa animal model. Intact (sham) versus castrated male Fisher-344 rats were evaluated in 2 scenarios: A) Carcinogenesis: After randomization to SHAM (n=5) and Castration (n=5), carcinogenesis was induced by four intravesical doses of 1.5 mg/kg n-methyl-n-nitrosourea (MNU) every 15 days. B) Treatment: After ultrasonographic confirmed MNU induced papillary BCa on week 8, rats were randomized to SHAM (n=5) and Castration (N=5) and offered 6 weekly intravesical treatment of 106 CFU of BCG in 0.2 ml saline. After 15 weeks of protocol, animals were euthanized and the urinary bladders submitted to histopathology, immunohistochemistry (IHC), and Western blotting (WB). Urothelial cell proliferation was measured by Ki-67 IHC and TLR-4 expression quantified by IHC and WB. RESULTS: Castration induced higher TLR-4 urothelial expression (p=.007) and anticarcinogenic effect with less urothelial tumors (60 vs. 80%) and lower urothelial cell proliferation compared to intact animals (p=.008). In the intravesical BCG treatment setting, castration has potentialized the BCG activation of TLR-4 (p=.007) with no residual in situ carcinoma compared to intact animals, suggesting the potential to amplify the immune initiation phase activated by BCG. CONCLUSIONS: To our knowledge, this is the first description of TLR-4 urothelial expression hormonal modulation. The described male immunocompetent animal model will help to improve the knowledge of urothelial cancer gender diversities and PRRs modulations with treatment implications. Source of Funding: none © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e199 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Leonardo Reis More articles by this author Expand All Advertisement PDF DownloadLoading ...

Identification of Morphological Features of Fibrillar Collagen Indicative of Tumor Progression in Bladder Cancer

ObjectiveBladder cancer is the most common malignancy involving the urinary system. As with all types of cancer, accurate monitoring of stage progression is critical in determining proper treatment strategies and predicting clinical outcomes. Collagen fibers are a major component of tumor microenvironment, and their post‐translational modifications during the course of tumor progression are well‐documented in a number of malignancies including bladder cancer. In the present feasibility study we sought to determine whether quantitative morphological features extracted from collagen fibers can be used to distinguish between urothelial carcinoma tumors pre‐muscle‐invasion (stage pT1) and post‐muscle‐invasion (stage pT2).MethodsPicroSirius Red (PSR) staining was used to detect fibrillar collagen in 8 tissue microarray (TMA) cores acquired from Biomax (4 stage pT1, 4 stage pT2). We applied auto‐fluorescence imaging techniques to detect the PSR signal, and employed the CT‐Fire freeware to automatically extract individual collagen fibers. Five morphological features (including length, width, straightness, orientation, and anisotropy) were computed for each detected fiber. Individual features were then combined within each slide to form five corresponding histograms per TMA core. Finally, for each histogram we computed two measures of central tendency (i.e., mean and median), two measures of spread (i.e., range and standard deviation), and two measures of symmetry (i.e., skewness and kurtosis).ResultsTo identify any statistically‐significant differences between the pT1 and pT2 groups, we employed the student’s t‐test (run in Python 3.8.5) to compare the two groups with respect to each of the five morphological features and across each of the six distributional measures mentioned in the Methods section (30 comparisons in total). Of these 30 comparisons, all but one (i.e., median) of the six straightness measures resulted in a p‐value &lt; 0.05. Further, each of the two measures of spread (i.e., range and standard deviation) of straightness returned a p‐value &lt; 0.001. We are currently working on expanding the size of our dataset to further validate these results.ConclusionOur preliminary findings are in line with the hypothesis that collagen remodeling is strongly associated with muscle‐invasive bladder cancer, and suggest that quantitative features measuring the straightness of individual fibers may be used to improve tumor staging. This type of quantitative approach is particularly helpful in distinguishing between stage pT1 and stage pT2 of bladder cancer, a task which can be challenging at times even for experienced pathologists.

Possible role of 5-alpha reductase inhibitors in non-invasive bladder urothelial neoplasm: multicenter study.

About 75% of urothelial bladder cancers are non-muscle invasive (NMIBC), and limited to mucosa (Ta or CIS) or sub-mucosa (T1). An increase of androgen expression and androgen receptors has a positive effect on oncogenic expression. We aimed to evaluate whether 5-alpha reductase inhibitors (5-ARI) have a role in NMIBC. We retrospectively evaluated the clinical and pathological data of 423 patients with NMIBC who underwent transurethral bladder resection. We considered the number of resections, number of total recurrences, time of recurrences, and histopathology details. The population was classified into two groups: treated and untreated with 5-ARIs. The enrolled patients were in treatment with 5ARIs for symptomatic prostatic hyperplasia for at least 12 months. Mean follow-up time was 30.43 months. Patients treated with 5-ARIs had a lower rate of recurrence (14%) than the untreated group (37%). There was a significant difference in the mean number of recurrences between the untreated and the treated group (P=0.006). Furthermore, the treated group showed a significantly greater number of low than high grade tumors, compared to the untreated group (P≤0.05). There was a significant decrease in the number of muscle invasive tumors in treated patients (P=0.032). The recurrence-free survival rate of patients treated with 5-ARIs was significantly higher (P=0.0001). Long-term treatment with 5-ARIs might reduce the risk of bladder tumor recurrence, extension of lesions and increase the recurrence-free survival rate. A long-term, randomized prospective study could definitively assess the possible role of these drugs.

Nephrogenic Adenoma Intermixed With Urothelial Carcinoma.

Nephrogenic adenoma (NA) is a common urinary tract lesion typically associated with urothelial disruption, leading to implantation of shed renal tubular cells. NA may demonstrate a spectrum of architectural and cytologic features mimicking urothelial carcinoma (UC), adenocarcinoma (including clear cell adenocarcinoma and prostatic adenocarcinoma), and invasion. However, admixed UC and NA has not been described. To describe cases where the NA was intimately intermixed with UC, potentially mimicking variant differentiation or invasion. In 3 health care systems we identified specimens of NA and UC intimately intermixed with each other where they could mimic a spectrum of one lesion. We assessed patterns of NA and clinical implications of misdiagnosing NA as glandular differentiation of UC. There were 4 women and 29 men (median age, 72 years; range, 31-89 years). Twenty-four patients had transurethral resections, 3 had biopsies, and 6 had major resections. Fourteen had noninvasive high-grade papillary UC, 6 had carcinoma in situ, and 11 had invasive high-grade UC, and in 2 patients, NA developed in a papillary urothelial neoplasm with extensive denudation. Three patients had fibromyxoid NA infiltrated by invasive UC. Classical NA (n = 30) had tubulopapillary (n = 18), pure tubular (n = 7), or pure papillary architecture (n = 5). In 1 lesion, NA was present in muscularis propria, and 2 involved adventitia. NA could have been misdiagnosed as invasion in 17 of 22 (77%) noninvasive tumors, or higher stage in 19 of 33 (58%). NA can be intermingled with high-grade UC, expanding the spectrum of entities that must be considered in the differential diagnosis, as it may mimic glandular or tubular differentiation, invasion, and a higher stage of disease. Misinterpretation of NA in such a setting may incorrectly convey a more aggressive biological potential of cancer to clinicians.

PD26-08 LOW-GRADE UROTHELIAL CARCINOMA RECURS AT A TEMPO THAT NATURALLY ACCELERATES FROM ADAGIO TO ALLEGRO

You have accessJournal of UrologyCME1 May 2022PD26-08 LOW-GRADE UROTHELIAL CARCINOMA RECURS AT A TEMPO THAT NATURALLY ACCELERATES FROM ADAGIO TO ALLEGRO Alex Sankin, Louise-Rae Cherrill, Rebecca H. Boucher, Maurice P. Zeegers, K. K. Cheng, Nicholas D. James, Mark Schoenberg, Ilir Agalliu, and Richard T. Bryan Alex SankinAlex Sankin More articles by this author , Louise-Rae CherrillLouise-Rae Cherrill More articles by this author , Rebecca H. BoucherRebecca H. Boucher More articles by this author , Maurice P. ZeegersMaurice P. Zeegers More articles by this author , K. K. ChengK. K. Cheng More articles by this author , Nicholas D. JamesNicholas D. James More articles by this author , Mark SchoenbergMark Schoenberg More articles by this author , Ilir AgalliuIlir Agalliu More articles by this author , and Richard T. BryanRichard T. Bryan More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002574.08AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Patients with low grade urothelial carcinoma are at a lifetime risk of recurrence and often require multiple interventions including repeat transurethral resections and intravesical therapy. We present an exploratory analysis of the frequency of recurrence in patients with low grade intermediate-risk non-muscle invasive bladder cancer (NMIBC) from a comprehensive large-scale prospective cohort in the UK. METHODS: The Bladder Cancer Prognosis Programme (BCPP) is a prospective cohort of 1191 patients with newly-diagnosed bladder cancer who were recruited from 2005-2011 at 10 urological centers within the West Midlands, UK, and who remain under follow-up. Risk group was assigned according to European Association of Urology criteria. Time to first recurrence is defined as the time from registration to detection of first recurrence of bladder cancer; time between first recurrence and second recurrence is defined as the time from the detection of the first recurrence to the detection of the second recurrence, and so on for time between subsequent recurrences. All recurrence-free intervals are calculated using the Kaplan-Meier method. RESULTS: We identified 379 patients with G1/G2 pTa tumors classified as intermediate risk (Table 1). Median age was 70 and 284/379 (75%) were male. 53% of patients had at least one recurrence after 5 years of follow-up. 1-year recurrence-free survival (RFS) was 75%, 2-year RFS was 61%, 3-year RFS was 54%, and 4-year RFS was 50%. The median time to 1st, 2nd, 3rd, 4th, and 5th sequential recurrence was 49, 19, 12, 14, and 10 months, respectively (Figure 1). CONCLUSIONS: Over half of patients with low grade urothelial tumors are destined to recur. As tumors recur sequentially over time, the rate of recurrence appears to accelerate. The natural cadence of tumor recurrence may represent a kinetic biomarker that could inform both surveillance and intervention protocols. Source of Funding: Urogen Pharma © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e490 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Alex Sankin More articles by this author Louise-Rae Cherrill More articles by this author Rebecca H. Boucher More articles by this author Maurice P. Zeegers More articles by this author K. K. Cheng More articles by this author Nicholas D. James More articles by this author Mark Schoenberg More articles by this author Ilir Agalliu More articles by this author Richard T. Bryan More articles by this author Expand All Advertisement PDF DownloadLoading ...

MiR-133b regulates the proliferation, colony formation, and invasion of bladder cancer cells via inhibiting SOX4.

Bladder cancer is one of the most common urothelial tumors with high incidence and mortality rates. Although it has been reported that microRNA (miR)-133b can regulate tumorigenesis of bladder cancer, the mechanism remains unclear. Sex-determining region Y-box transcription factor 4 (SOX4) exhibits an important role in tumorigenesis, but it is unclear whether SOX4 and miR-133b are associated with regulation of pathogenesis of bladder cancer. This study aims to determine the expressions of SOX4 and miR-133b in bladder cancer tissues and cells, investigate their effects on the proliferation, colony formation, and invasion of bladder cancer cells, and to explore the association between miR-133b and SOX4 in regulating biological featurss of bladder cancer cells. The bladder cancer and adjacent tissue samples of 10 patients who underwent surgical resection in the Second Xiangya Hospital of Central South Universty from Januray to June 2015 were obtained. The levels of miR-133b were tested by real-time PCR, and the protein levels of SOX4 were evaluated using Western blotting in bladder cancer tissues, matched adjacent tissues, and cell lines. The correlation between miR-133b expression and SOX4 expression in bladder cancer tissues was analyzed. Using the online database TargetScan, the relationship between SOX4 and miR-133b was predicted. MiR-133b mimics, miR-133b inhibitor, and short hairpin RNA (shRNA)-SOX4 were transfected into T24 cells by Lipofectamine 2000. The relationship between miR-133b and SOX4 was also verified by a dual-luciferase reporter assay. The proliferation of T24 cells cultured for 0, 12, 48, 72, and 96 h was evaluated by cell counting kit-8 (CCK-8) assay. The colony formation capacity of bladder cancer cells was tested after 14-day culture, and cell invasion capacity was evaluated with Transwell invasion assay. Bladder cancer tissue and bladder cancer cells had low level of miR-133b but high level of SOX4, compared with matched adjacent tissues and normal bladder epithelial cells. A negative correlation between miR-133b mRNA and SOX4 protein levels in bladder cancer tissues was also found (r=-0.84). The results of online database TargetScan showed that miR-133b targets at SOX4, and overexpression of miR-133b significantly attenuated the expression of SOX4 in T24 cells. Both overexpression of miR-133b and knockdown of SOX4 significantly inhibited the proliferation, colony formation, and invasion capacity of bladder cancer cells in vitro. SOX4 down-regulation restored the effects of miR-133b inhibitor on the proliferation, colony formation, and invasion capacity of T24 cells. The up-regulation of SOX4 contributes to the progression of bladder cancer, and miR-133b can regulate the proliferation, colony formation, and invasion of bladder cancer cells via inhibiting SOX4.

Characterization of the Peri-Membrane Fluorescence Phenomenon Allowing the Detection of Urothelial Tumor Cells in Urine.

Simple SummaryTo detect bladder cancer (BC), urinary cytology and cystoscopy are the primary diagnostic tests used. Urine cytology is non-invasive, easy to collect, with medium sensitivity and high specificity. It is an effective way to detect high-grade BC, but it is less effective on low-grade BC because the rate of equivocal results is much higher, making them difficult to detect. Despite the implementation of new diagnostics, urinary cytology and cystoscopy remain the gold standard. Instead of looking for new diagnostics, one of the new research areas is the improvement of urinary cytology. Recently, the fluorescent properties of plasma membranes of urothelial tumor cells, called peri-membrane fluorescence, found in urinary cytology have been shown to be useful in improving the early detection of BC. The main objective of this study was to characterize the peri-membrane fluorescence allowing the detection of urothelial tumor cells in urine.Urine cytology is non-invasive, easy to collect, with medium sensitivity and a high specificity. It is an effective way to detect high-grade bladder cancer (BC), but it is less effective on low-grade BC because the rate of equivocal results is much higher. Recently, the fluorescent properties of plasma membranes of urothelial tumor cells (UTC) found in urine cytology have been shown to be useful in improving the early detection of BC. This phenomenon is called peri-membrane fluorescence (PMF). Based on previous studies that have identified the PMF on UTCs, the main objective was to characterize this phenomenon. For this study, a software was specially created to quantify the PMF of all tested cells and different treatments performed. PMF was not found to be a morphological and discriminating feature of UTCs, all cells in shape and not from urine show PMF. We were able to highlight the crucial role of plasma membrane integrity in the maintenance of PMF. Finally, it was found that the induction of a strong cellular stress induced a decrease in PMF, mimicking what was observed in non-tumor cells collected from urine. These results suggest that PMF is found in cells able to resist this stress, such as tumor cells.

Dual-Energy CT applications in urinary tract cancers: an update.

Urothelial tumours are the fourth most common cancer in the world and account for the majority of tumours involving the bladder. The symptom that often leads to diagnosis is the presence of haematuria. Diagnosis is made by cystoscopy, which is currently the gold standard in bladder cancer. Computed tomography (CT) performed with pre- and post-contrastographic phases is essential in order to assess the loco-regional and distant extension of disease. The diagnosis and staging of upper tract urothelial cancer (UTUC) are best done with computed tomography urography and flexible ureteroscopy (URS). In the acquisition protocol of this type of tumour, a urographic phase is mandatory, which allows for an accurate diagnostic assessment of the renal pelvis, ureter and bladder, especially in papillary forms. The use of multiple acquisition phases, especially in this type of patient who will have to perform follow-up CTs, leads to the problem of overexposure to ionising radiation, as well as the frequent administration of iodinated contrast medium. For this reason, in recent year, the focus has been put on advanced technologies such as dual-energy CT (DECT), that is a method that can offer some advantages for both radiologist and patient, in the diagnosis of cancer and, in particular, urinary tract disease.

Moving translational mass spectrometry imaging towards transparent and reproducible data analyses: a case study of an urothelial cancer cohort analyzed in the Galaxy framework

BackgroundMass spectrometry imaging (MSI) derives spatial molecular distribution maps directly from clinical tissue specimens and thus bears great potential for assisting pathologists with diagnostic decisions or personalized treatments. Unfortunately, progress in translational MSI is often hindered by insufficient quality control and lack of reproducible data analysis. Raw data and analysis scripts are rarely publicly shared. Here, we demonstrate the application of the Galaxy MSI tool set for the reproducible analysis of a urothelial carcinoma dataset.MethodsTryptic peptides were imaged in a cohort of 39 formalin-fixed, paraffin-embedded human urothelial cancer tissue cores with a MALDI-TOF/TOF device. The complete data analysis was performed in a fully transparent and reproducible manner on the European Galaxy Server. Annotations of tumor and stroma were performed by a pathologist and transferred to the MSI data to allow for supervised classifications of tumor vs. stroma tissue areas as well as for muscle-infiltrating and non-muscle infiltrating urothelial carcinomas. For putative peptide identifications, m/z features were matched to the MSiMass list.ResultsRigorous quality control in combination with careful pre-processing enabled reduction of m/z shifts and intensity batch effects. High classification accuracy was found for both, tumor vs. stroma and muscle-infiltrating vs. non-muscle infiltrating urothelial tumors. Some of the most discriminative m/z features for each condition could be assigned a putative identity: stromal tissue was characterized by collagen peptides and tumor tissue by histone peptides. Immunohistochemistry confirmed an increased histone H2A abundance in the tumor compared to the stroma tissues. The muscle-infiltration status was distinguished via MSI by peptides from intermediate filaments such as cytokeratin 7 in non-muscle infiltrating carcinomas and vimentin in muscle-infiltrating urothelial carcinomas, which was confirmed by immunohistochemistry. To make the study fully reproducible and to advocate the criteria of FAIR (findability, accessibility, interoperability, and reusability) research data, we share the raw data, spectra annotations as well as all Galaxy histories and workflows. Data are available via ProteomeXchange with identifier PXD026459 and Galaxy results via https://github.com/foellmelanie/Bladder_MSI_Manuscript_Galaxy_links.ConclusionHere, we show that translational MSI data analysis in a fully transparent and reproducible manner is possible and we would like to encourage the community to join our efforts.

Clinicopathologic Spectrum of Secondary Solid Tumors of the Prostate of Nonurothelial Origin: Multi-institutional Evaluation of 85 Cases.

Secondary involvement of the prostate by urothelial or hematolymphoid neoplasms is relatively common and well-described. In contrast, less is known about the clinicopathologic spectrum of secondary solid tumors of the prostate of nonurothelial origin. This study evaluated a series of secondary nonurothelial solid tumors of the prostate diagnosed at 21 institutions. Eighty-five patients with a median age at diagnosis of 64 years were included. Sixty-two patients had clinically manifest disease (62/85, 73%), 10 were diagnosed incidentally (10/85, 12%), and 13 (13/85, 15%) had no detailed clinical data available about symptomatology at presentation. Among patients with clinically manifest disease, the most common symptoms and signs were lower urinary tract symptoms (either obstructive of irritative; 36/62, 58%), abdominal or pelvic pain or discomfort (16/62, 26%), and hematuria (12/62, 19%). Metastasis and direct invasion occurred at roughly similar frequencies (47% vs. 42%) in this series, and in 11% of the cases, the mechanism of spread to the prostate was unclear/uncertain. Overall, among tumors with confirmed sites of origin, the most common primary sites were gastrointestinal tract (53/85, 62%), lung (9/85, 11%), skin (6/85, 7%), and testis (4/85, 5%). Among metastases, the most common tumor types were lung carcinomas (9/40, 23%), colorectal adenocarcinomas (7/40, 18%), melanoma (6/40, 15%), and germ cell tumors (6/40, 15%). This study demonstrated that secondary involvement of the prostate by solid tumors of nonurothelial origin is commonly symptomatic and that the most frequent sites of origin are the gastrointestinal tract, lung, skin, and testis. These findings are worth considering when lesions with unusual cytomorphology and/or architecture are encountered in prostate specimens.

The Diagnostic Dilemma of Urothelial Tissue Fragments in Urinary Tract Cytology Specimens.

Since the release of The Paris System for Reporting Urinary Cytology (TPS), the assessment of urine cytology specimens has primarily focused on the detection of high-grade urothelial carcinoma (HGUC) and carcinoma in situ (CIS). Fortunately, the malignant cells in these lesions tend to be loosely cohesive, resulting in the natural exfoliation of individual malignant cells into the urine. However, HGUC/CIS lesions occasionally exfoliate larger fragments which can be difficult to assess due to cellular overlap and fragment three-dimensionality. Furthermore, reactive benign urothelial fragments and fragments from low-grade urothelial neoplasms (LGUN) may also be seen in urine specimens and contain atypical cytomorphologic features. As a result, the significance of urothelial tissue fragments (UTFs) is often unclear. Herein, we discuss the literature on UTFs before and after the implementation of TPS, as well as strategies to help overcome this diagnostic challenge.

Utility of reporting urine cytology samples as per the Paris system

Background: Urinary tract cytology is an accurate test for the detection of urothelial malignancy especially high-grade urothelial carcinoma. The Paris System for Reporting Urinary Cytology was introduced to standardize urinary tract cytology reporting. We aim to evaluate the utility of reporting urinary cytology as per this system and correlate with histopathology. Materials and Methods: This is a descriptive cross-sectional prospective study conducted on urine samples submitted for cytological examination at Kathmandu Medical College Teaching Hospital, Sinamangal, Nepal between 1st November 2020 to 31st July 2021. Ethical consent was taken from the Institutional Review Committee. Urine cytology was reported as per The Paris System for Reporting Urinary Cytology and correlated with the histopathologic diagnosis. Sensitivity, specificity, positive predictive value, and negative predictive value were calculated for the high-grade and low-grade urothelial lesion. Results: A total of 104 urine samples were evaluated. Biopsy specimens were available for 38 cases. Urine cytology consisted of 1.92% non-diagnostic cases, 69.23% negative for high-grade urothelial carcinoma, 5.76% atypical urothelial cells, 5.76% suspicious for high-grade urothelial carcinoma, 13.46% high-grade urothelial carcinoma, and 3.84% low-grade urothelial neoplasm respectively. Sensitivity, specificity, negative predictive value and positive predictive value were 90.48%, 82.35%, 86.37% and 87.5% respectively for high grade urothelial carcinoma and 40%, 93.9%,50% and 91.17% respectively for low grade urothelial lesions. Conclusions: Our study shows that reporting urine cytology as per The Paris System for Reporting Urinary Cytology provides high sensitivity for the detection of high-grade urothelial lesions.

Phase 1 Clinical Trial Evaluating the Safety and Anti-Tumor Activity of ADP-A2M10 SPEAR T-Cells in Patients With MAGE-A10+ Head and Neck, Melanoma, or Urothelial Tumors.

BackgroundADP-A2M10 specific peptide enhanced affinity receptor (SPEAR) T-cells are genetically engineered autologous T-cells that express a high-affinity melanoma-associated antigen (MAGE)-A10-specific T-cell receptor (TCR) targeting MAGE-A10-positive tumors in the context of human leukocyte antigen (HLA)-A*02. ADP-0022-004 is a phase 1, dose-escalation trial to evaluate the safety and anti-tumor activity of ADP-A2M10 in three malignancies (https://clinicaltrials.gov: NCT02989064).MethodsEligible patients were HLA-A*02 positive with advanced head and neck squamous cell carcinoma (HNSCC), melanoma, or urothelial carcinoma (UC) expressing MAGE-A10. Patients underwent apheresis; T-cells were isolated, transduced with a lentiviral vector containing the MAGE-A10 TCR, and expanded. Patients underwent lymphodepletion with fludarabine and cyclophosphamide prior to receiving ADP-A2M10. ADP-A2M10 was administered in two dose groups receiving 0.1×109 and >1.2 to 6×109 transduced cells, respectively, and an expansion group receiving 1.2 to 15×109 transduced cells.ResultsTen patients (eight male and two female) with HNSCC (four), melanoma (three), and UC (three) were treated. Three patients were treated in each of the two dose groups, and four patients were treated in the expansion group. The most frequently reported adverse events grade ≥3 were leukopenia (10), lymphopenia (10), neutropenia (10), anemia (nine), and thrombocytopenia (five). Two patients reported cytokine release syndrome (one each with grade 1 and grade 3), with resolution. Best response included stable disease in four patients, progressive disease in five patients, and not evaluable in one patient. ADP-A2M10 cells were detectable in peripheral blood from patients in each dose group and the expansion group and in tumor tissues from patients in the higher dose group and the expansion group. Peak persistence was greater in patients from the higher dose group and the expansion group compared with the lower dose group.ConclusionsADP-A2M10 has shown an acceptable safety profile with no evidence of toxicity related to off-target binding or alloreactivity in these malignancies. Persistence of ADP-A2M10 in the peripheral blood and trafficking of ADP-A2M10 into the tumor was demonstrated. Because MAGE-A10 expression frequently overlaps with MAGE-A4 expression in tumors and responses were observed in the MAGE-A4 trial (NCT03132922), this clinical program closed, and trials with SPEAR T-cells targeting the MAGE-A4 antigen are ongoing.