Cases Of High-grade Urothelial Carcinoma Research Articles

Comparison of tissue biomarkers between Non-Schistosoma and Schistosoma-Associated Urothelial Carcinoma

BackgroundHigh-grade urothelial carcinoma either non-Schistosoma (NS-UBC) or Schistosoma (S-UBC)-associated is the tenth cause of death worldwide and represents a serious therapeutic problem. AimEvaluation of the immmunohistochemical expression of tumor necrosis factor-alpha (TNFα), epidermal growth factor receptor (EGFR), programmed cell death protein-1 (PDL1), estrogen receptor-alpha (ERα) and UroplakinIII, in the high-grade in NS-UBC and S-UBC as potential prognostic and therapeutic targets analyzed through estimation of area percentage, optical density and international pathological scoring system for each marker. Material and methodsSixty high grade urothelial carcinoma cases were enrolled in the study (30 cases of NS-UBC and 30 cases of S-UBC). The cases were immunohistochemically-assessed for TNFα, EGFR, PDL1, ERα and Uroplakin III expression. In S-UBC, parasite load was also evaluated for correlation with the immunohistochemical markers’ expression in S-UBC. ResultsThe area percentage of immune-expression of TNFα and EGFR was higher in S-UBC compared to NS-UBC. On the other hand, the NS-UBC displayed statistically-higher expression of PDL1 and uroplakinIII (p-value <0.001). ERα revealed higher, yet, non-significant expressions in S-UBC compared to NS-UBC (p-value =0.459). PDL1 expression showed the most superior record regarding area percentage (64.6± 34.5). Regarding optical density, TNF-α showed the highest transmittance expression (2.4 ± 0.9). EGFR positively correlated with PDL1 in S-UBC (r= 0.578, p-value =0.001) whereas in NS-UBC, TNFα and PDL1 (r=0.382, p-value=0.037) had positive correlation. Schistosoma eggs in tissues oppose uroplakin III expression and trigger immunomodulation via PDL1. ConclusionDue to lower UroplakinIII expression, S-UBC is supposed to have a poorer prognosis. Hormonal therapy is not hypothesized due to a very minimal ERα expression in both NS-UBC and S-UBC. Regarding immunotherapy, anti-TNF-α is suggested for S-UBC whilst in NS-UBC, blockading PDL1 might be useful. Targeted EGFR therapy seems to carry emphasized outcomes in S-UBC. Correlations encourage combined immune therapy in NS-UBC; nevertheless, in S-UBC, combined anti-EGFR and PDL1 seem to be of benefit.

P53 and vimentin double immunostaining to differentiate between high-grade urothelial carcinoma cells and benign atypical cells

Introduction: Urine cytology is an indispensable test for detecting high-grade urothelial carcinoma (HGUC); however, the distinction between HGUC cells and morphologically similar benign atypical cells poses clinical challenges. In this study, we performed double immunostaining for p53 and vimentin to establish a diagnostic method to accurately distinguish HGUC cells from benign atypical cells. Methods: This study included 41 cases of HGUC, 11 of urolithiasis, and 22 of glomerular disease diagnosed histopathologically or clinically. After preparing urine cytology specimens from voided urine samples, p53 immunostaining was performed, and the p53 positive intensity and p53 positivity rate were calculated. Subsequently, vimentin immunostaining was performed on the same specimens to calculate the rate of vimentin positivity. Results: The HGUC cell group had a mean p53 positive intensity of 2.40, a mean p53 positivity rate of 73.2%, and a mean vimentin positivity rate of 5.1%. In contrast, the mean p53 positive intensity, p53 positivity rate, and vimentin-positivity rate was 1.63, 36.7%, and 66.2%, respectively, in the benign atypical cell group. There were significant differences between the two groups for each parameter. Moreover, two multiple logistic regression models combining the results of these three parameters exhibited higher sensitivity and specificity than solely assessing the p53 positive intensity, positivity rate, and vimentin positivity rate. Conclusion: Since double immunostaining with p53 and vimentin distinguishes HGUC cells from benign atypical cells, it could be used to improve the diagnostic accuracy of urine cytology.

Mixed-Grade Urothelial Carcinoma: Insights Into Clinical Behavior and Prognostic Implications Compared to Pure Low-Grade and High-Grade Urothelial Carcinomas.

Low-grade urothelial carcinoma (LGUC) and high-grade urothelial carcinoma (HGUC) are distinguished based on architectural and cytological features, with the anticipation that HGUC exhibits more aggressive behavior and a worse prognosis compared to LGUC. The current World Health Organization classification recognizes mixed-grade urothelial carcinoma (MGUC, for the purposes of this study) as a separate category that behaves like LGUC if the high-grade component is <5% and states that any tumor with ≥5% high-grade component should be graded as HGUC. To evaluate the risk of tumor recurrence, grade, and stage progression of MGUC compared to LGUC and HGUC. A total of 150 de novo noninvasive polypoid urothelial carcinomas (41 cases of MGUC, 59 of LGUC, and 50 of HGUC) were included. Tumor recurrence, grade, and stage progression were compared among the MGUC, LGUC, and HGUC cases. Tumor recurrence was observed in 14 of 41 (34.2%) of MGUC, 33 of 59 (55.9%) of LGUC, and 28 of 50 (56%) of HGUC. Grade progression occurred in 5 of 41 (12.2%) of MGUC cases and 5 of 59 (8.5%) of LGUC cases. No stage progression was observed in LGUC or MGUC cases, while 7 of 50 (14%) of HGUC cases showed stage progression. MGUC was associated with lower odds and hazard of recurrence compared to LGUC. The rate of grade progression was higher in MGUC and occurred after a shorter interval compared to LGUC. MGUC showed a prognosis closer to LGUC. Our study supports the current recommendation to classify tumors with <5% high-grade component as MGUC, as these tumors display clinical characteristics and outcomes close to that of pure LGUC.

Investigation of Ki67 and Phospho-Histone H3 Expressions in Urothelial Carcinoma of the Bladder by Immunohistochemical Method.

In our study, it is aimed to investigate the relationship between Ki67 and phospho-histone H3 (pHH3) expressions in bladder urothelial carcinomas, with clinicopathological parameters and survival, which have prognostic value. The study included 44 cases of high-grade urothelial carcinoma (HGUC), 37 cases of low-grade urothelial carcinoma (LGUC), and 11 nontumoral bladder cases. Ki67 and pHH3 were applied to the paraffin blocks of the tissues of 81 urothelial carcinoma and 11 nontumoral bladder cases by immunohistochemical method.Percentages of Ki67 and pHH3 expressions were evaluated by digital imaging analysis method. Expression percentages were compared with various clinicopathological parameters, and the relationship between them was evaluated. Ki67 was expressed in 28% of urothelial carcinoma cases and 1% of nontumoral cases. pHH3 was expressed in 10.32% of urothelial carcinoma cases and 0.16% of nontumoral cases. In our study, we found significantly higher Ki67 and pHH3 expressions in urothelial carcinoma compared to nontumoral cases. There was a statistically significant relationship (p < 0.05) and a positive correlation between Ki67 expression and lymphovascular invasion, pT stage, and histological grade. A statistically significant relationship (p < 0.05) and a positive correlation were found between pHH3 expression and lymphovascular invasion, pT stage, recurrence, and histological grade. In addition, a statistically significant relationship was found between Ki67 and pHH3 expressions. In our study, survival was found to be low in high-grade urothelial carcinoma cases with lymphovascular invasion, advanced age (65 years and older), and high Ki67 and pHH3 expression rates. According to our findings, high Ki67 and pHH3 expressions were found to be associated with poor prognostic parameters such as advanced pathologic stage, high histologic grade, and low survival.Our findings suggest that Ki67 and pHH3 may play a role in the differentiation, progression, and aggressive behavior of urothelial carcinoma. However, further studies are needed to confirm our findings and determine the role of these markers in urothelial carcinoma.

A Case Report: High-Grade Urothelial Carcinoma in the Renal Pelvis with Complete Kidney and Ureter Duplication Featuring Heterologous Differentiation

This report describes a considerably rare case of high-grade urothelial carcinoma of the renal pelvis and ureter, presenting with heterologous differentiation, in a patient with bilateral duplicated kidneys. A 73-year-old male experienced intermittent gross hematuria for 5 months, accompanied by lower back and abdominal pain. Ultrasound and computed tomography scans revealed bilateral renal and ureteral duplication with multiple tumors in the left renal pelvis. A total nephroterectomy and bladder cuff resection were performed on the left two nephrons. Multiple space-occupying lesions were identified in the left renal pelvis and ureter. Histopathological examination showed poorly differentiated and diverse tumor cells, manifesting as sarcomatoid carcinoma, papillary adenocarcinoma, and infiltrating high-grade urothelial carcinoma. The tumor infiltrated the subcutaneous fibrous connective tissue of the renal pelvis and the full thickness of the ureter. Given the rarity of recurrent renal urothelial carcinoma with heterogeneous differentiation, comprehensive imaging and pathological assessments are vital to delineate the nature of the lesion and the direction of tissue pathological heterologous differentiation. These evaluations guide early radical surgical interventions, improving survival rates.

Are we on track for diagnosing high-grade urothelial carcinoma with a minimum quantity of five malignant cells in lower tract specimens? Critical analysis of The Paris System Quantitation Criteria.

The Paris System for Reporting Urinary Cytology (TPS) has gained universal acceptance as the standard for reporting urine cytology requiring at least 5-10 malignant cells to diagnose high-grade urothelial carcinoma (HGUC) in lower and upper urinary tract specimens, respectively. These quantitation criteria are still subject to discussion, and this study specifically aims to validate the quantitation criterion of HGUC in lower urinary tract. The authors reviewed two cohorts of lower urinary tract cases. The first cohort consisted of 100 liquid-based ThinPrep slides with the diagnosis of HGUC having positive histology on concurrent or follow-up biopsies within 3months. The second cohort was 36 HGUC cases with negative histology on concurrent biopsies and within 3months. The number of high-grade cells (HGCs) meeting the TPS qualitative criteria were counted under the light microscope driven in a grid-like manner. The first 100 urine samples showed five cases (5.0%) with three HGCs, three cases (3.0%) had four HGCs, five cases (5.0%) showed five HGCs, and 25 cases (25.0%) had between 6-10 HGCs. The risk of high-grade malignancy (ROHM) in cases with five or more HGCs was 100%, whereas those with three HGCs was 60.0%. The second cohort of HGUC was considered "positive" despite a negative histology. This study confirms that quantitation is an essential key to diagnose HGUC. The current TPS criterion of a minimum of five malignant cells in lower tract is robust with a ROHM of 100%. Diagnosing HGUC with less than five HGCs runs the risk of lowering the ROHM.

CD105 and CD34 as Markers of Angiogenesis of Urothelial Carcinoma: A Cross Sectional Study

Background Urothelial carcinoma (UC) of the bladder is a major cause of morbidity and mortality throughout the world. The present study was carried out to see the expression of CD105 and CD34 as marker of angiogenesis in UC and study their relationship with grades and muscle invasion. Methods 50 cases of histopathologically diagnosed UC were included. CD105 and CD34 IHC staining was performed. Microvessel density (MVD) evaluation by CD105 and CD34 was compared. The number of microvessels were counted in high power in three hot spots. The highest value obtained amongst three fields were reported as MVD. Statistical analysis was done by (SPSS 28.00 version for windows) using independent t-test. Results There were 45 high grade cases UC 5 low grade cases of UC. Invasive papillary UC was seen in 25 cases (50%) followed by invasive UC seen in 13 cases (26%). Both Lamina propria (LP) invasion and muscularis propria invasion were noted in 16 cases of high-grade urothelial carcinoma. Only LP invasion was seen in 3 cases of low grade and 25 cases of high-grade UC respectively. Evaluation of MVD CD105 and MVD CD34 did not show any significant association of MVD with grade with a p value of 0.214 &amp; 0.429 respectively. Aso there was no significant association of MVD CD105 (p=0.580) &amp; CD34 (p=0.747) with muscularis propria invasion. Overall mean CD105 and CD34 were similar with no statistical difference between the two markers. Conclusion Both CD105 and CD34 can be used as marker to assess angiogenesis. However, there is no significant association of MVD with grade or muscle invasion in UC.

The preliminary efficacy of "quadri-combination" therapy on the treatment of bladder cancer

To investigate the safety and efficacy of "quadri-combination" therapy including maximal transurethral resection of bladder tumor (mTURBT), combined with systemic chemotherapy and immunotherapy, concurrent radiotherapy, and immune maintenance therapy. The clinical data of 8 patients with bladder cancer who could not tolerate or refused radical cystectomy at the Department of Urology, Peking University Cancer Hospital from November 2019 to October 2021 were retrospectively analyzed. There were 5 males and 3 females with a mean age of 69 years. The Eastern Cooperative Oncology Group(ECOG) score was 0 in 6 cases and 1 in 2 cases. There were 5 cases of high-grade urothelial carcinoma (1 case of T3b; 2 cases were T2; 2 cases of T1 stage, with multiple tumors and repeated recurrence), 1 case of high-grade urothelial carcinoma with carcinoma in situ (T1/Tis stage), 1 case of high-grade urothelial carcinoma with squamous differentiation (T3b stage), and 1 case of high-grade urothelial carcinoma with glandular differentiation (T2). All patients underwent "quadri-combination" therapy.The patient's tolerance, success rate of bladder preservation and prognosis were evaluated.The median follow-up time was 22.5 (12-35) months. One patient with high-grade muscle-invasive bladder cancer (T2) received mTURBT, albumin-bound paclitaxel and durvalumab combined therapy for 3 cycles, concurrent radiotherapy, and immune maintenance therapy for 18 months, and the tumor recurrence was found. The pathology was high-grade urothelial carcinoma. Salvage radical cystectomy combined with pelvic lymph node dissection is recommended. The remaining 7 patients were regularly reexamined, and no recurrence or metastasis was found.The 2-year progression-free survival rate was 80%, and the success rate of bladder preservation was 87.5%(7/8). Treatment-related adverse reactions were resolved by symptomatic treatment, and patients' compliance and tolerance were acceptable.The "quadri-combination" bladder-preserving therapy is feasible and well tolerated, but further studies are needed.

Cyto-histo correlation and false-negative urine: Before and after the Paris system for reporting urinary cytology.

The impact of implementing the Paris system (TPS) on the rate of discrepant cases in the negative for high-grade urothelial carcinoma (NHGUC) category that had a subsequent diagnosis of high-grade urothelial carcinoma (HGUC) on histology is not well studied. We adopted TPS in May 2019. We searched discrepant cases with negative urine cytology 2017-2019 in our cyto-histo correlation database. The urine cytology and follow-up biopsy/resection were reviewed by a cytopathologist who also did Genitourinary (GU) Pathology subspecialty sign-out. Voided urine and instrumented urine were included in this study. There were total of 70 discrepant cases with negative cytology interpretation but HGUC on the subsequent biopsy or resected specimen. Following the TPS criteria, the rate of discrepant negative cytology cases increased from 6 cases between January 2017 and May 2019 to 64 cases after May 2019 when we adopted TPS. There were 2 discrepant negative cases in 2017, 3 cases in 2018, and 65 cases in 2019. Out of 65 cases in 2019, 64 cases were identified after May 2019. Additional 55 urine cytology slides were reviewed according to the TPS criteria, of which, the diagnoses remained unchanged in 45 (82%) cases and 10 (19%) cases were reassigned to either atypical or suspicious categories. The discrepancy was noted more on the instrumented urine and the upper tract urine. However, the false-negative rate rose faster in voided urine and lower tract urine. The risk of HGUC with the category of NHGUC was 0.03% in 2017, 0.05% in 2018, and 1.06% in 2019 at our institution. The increase in false-negative rate could not be attributed to a single cytopathologist. After adopting TPS for reporting urine cytology, there was an increase in HGUC from negative urine cytology which was subsequently confirmed on histology as cases of HGUC. The quality control of negative urines could be important monitoring the process when implementing TPS.

Evaluating the role of Z-stack to improve the morphologic evaluation of urine cytology whole slide images for high-grade urothelial carcinoma: Results and review of a pilot study.

Whole slide imaging (WSI) adoption has been slower in cytopathology due, in part, to challenges in multifocal plane scanning on 3-dimensional cell clusters. ThinPrep and other liquid-based preparations may alleviate the issue by reducing clusters in a concentrated area. This study investigates the use of Z-stacked images for diagnostic assessment and the experience of evaluating urine ThinPrep WSI. Thirty ThinPrep urine cases of high-grade urothelial carcinoma (n=22) and cases of negative for high-grade urothelial carcinoma (n=8) were included. Slides were scanned at 40× magnification without Z-stack and with Z-stack at 3 layers, 1 μm each. Six cytopathologists and 1 cytotechnologist evaluated the cases in 2 rounds with a 2-week wash-out period in a blinded manner. A Cohen's Kappa (CK) calculated concordance rates. A survey after each round evaluated participant experience. CK with the original report ranged from 0.606 to 1.0 (P < .05) without Z-stack and 0.533 to 1.0 (P < .05) with Z-stack both indicating substantial-to-perfect concordance. For both rounds, interobserver CK was moderate-to-perfect (0.417-1.0, P < .05). Intraobserver CK was 0.697-1.0 (P < 0.05), indicating substantial to perfect concordance. The average scan time and file size for slides without Z-stack and with Z-stack are 6.27 minute/0.827 GB and 14.06 minute/2.650 GB, respectively. Surveys demonstrated a range in comfort and use with slightly more favorable opinions for Z-stacked cases. Z-stack images provide minimal diagnostic benefit for urine ThinPrep WSI. In addition, Z-stacked urine WSI does not justify the prolonged scan times and larger storage needs compared to those without Z-stack.

A surgical case of high-grade urothelial carcinoma of the renal pelvis complicated with giant hydronephrosis, giving rise to diagnostic difficulties on a cytological examination

BackgroundWe report a surgical case of urothelial carcinoma of the renal pelvis, resulting in diagnostic difficulties on cytological examination.Case presentationA man in his late 70s underwent nephrectomy for giant hydronephrosis and renal cysts after nephrostomy and renal cyst puncture and drainage. On all cytological examinations performed before surgery, including nephrostomy urine, renal cyst fluid, catheterized bladder urine, and bladder washings, we were unable to make any conclusive diagnosis of malignancy. The pathological diagnosis of the surgical specimen concluded that this was a case of high-grade urothelial carcinoma of the renal pelvis with focal squamous differentiation (pT4). Liver and lung metastases were identified 3 months after surgery, and the patient died 2 months later.ConclusionIt was very difficult to make a conclusive diagnosis using cytological specimens because of the presence of a small number of atypical cells with severe degenerative changes. Since clinicians cannot predict the potential for malignancy on preoperative imaging findings, it is critical to consider the difficulties in clinically making a correct diagnosis of urothelial carcinoma of the upper urinary tract, especially in cases complicated with giant hydronephrosis.

EXPRESSION OF OCT-4 STEM CELL MARKER IN URINARY BLADDER CANCER AND ITS CORRELATION WITH INVASIVENESS

This study’s objective is to evaluate the expression of Octamer-binding transcription factor 4 (OCT-4) as a prognostic biomarker for urinary bladder cancer (UBC) along with its histological grade and tissue invasiveness. It is also called the POU domain, class 5, transcription factor I(POU5FI). OCT-4 protein is a transcription factor and a well-known marker of cancer stem cells. This study aimed to validate the OCT-4 value in urinary bladder cancer. FFPE tissue blocks from ten patients of different grades and invasiveness of urothelial carcinoma were studied and correlated with the degree of OCT-4 expression by immunohistochemistry. The mean histochemical expression score in seven high-grade urothelial carcinoma cases was 8.4 as compared to 5.0 in three low-grade urothelial carcinoma cases. Similarly, the mean histochemical expression score in seven lamina invasive urothelial carcinoma was 5.9 as compared to 7.5 in detrusor muscle-invasive urothelial carcinoma. There was a positive correlation between OCT-4 expression and high grade and invasive urinary bladder cancer.

Molecular stratification of high-grade urothelial carcinoma by immunohistochemistry with its histomorphological and clinical correlation.

Urothelial carcinoma poses a significant cause of morbidity and mortality. The recent classification of Tumors of Urinary System by World Health Organization fourth edition) has elucidated its molecular subtypes and its associated prognostic significance. We used immunohistochemistry marker expression (CK5/6, CK20, CD44, EGFR) as a surrogate marker, to stratify 150 cases of high-grade urothelial carcinoma into the intrinsic molecular subtypes. A correlation was also done with immunohistochemical markers p53, p21, E-cadherin and Ki-67. On subtyping, 47.3% cases were basal, 42.7% luminal and 10% remained unclassified. We did not find GATA3 useful for molecular stratification in our study. Muscle invasion was seen in 59% of basal and 31% of luminal subtype (P = 0.016). Squamous differentiation was most commonly associated with basal subtype (P < 0.001). EGFR expression was seen in 62% of basal and 38% of luminal subtype (P = 0.014), and thus can be used as an additional marker for molecular stratification. Overexpression of p53 was seen in 64% cases of muscle invasive and 36% of non-muscle invasive high-grade carcinomas (P < 0.0001). An inverse relationship was observed between p53 and p21 immunoexpression (r = -0.494) (P < .0001). The overall survival at 1- and 2-year interval was more in the luminal subtype, suggesting an early mortality in basal group, (P = 0.827), and at 6 years both the groups had almost similar results. High-grade urothelial carcinoma is challenging in terms of therapeutic strategy. Increased understanding of underlying molecular basis helps identifying targetable treatment options, and newer biomarkers will enhance predictive and prognostic stratification.

Digital image analysis of high-grade urothelial carcinoma in urine cytology confirms chromasia heterogeneity and reveals a subset with hypochromatic nuclei and another with extremely dark or "India ink" nuclei.

The Paris System for Reporting Urinary Cytology (TPS) uses hyperchromasia as major diagnostic criterion for high-grade urothelial carcinoma (HGUC). The purpose of the study was to evaluate cases that were diagnosed as HGUC by TPS and determine whether there are different chromatin distribution patterns (ie, subsets). Digital image annotations were performed on microscopic images of HGUC urine specimens with surgical biopsy/resection follow-up. Median gray values were generated for each cell. Neutrophils (polymorphonuclear leukocyte [PMN]) were also enumerated in each case to serve as an internal control. A HGUC/PMN ratio was generated for each case, and the cases were distributed. Sixty-nine HGUC cases yielded 2660 cells, including 2078 HGUC (30.1 cells/case) and 582 PMNs (8.4 cells/case). The average median gray value of an HGUC was 50.6 and of a PMN was 36.8 (P < .0001). Eight of 69 cases (11.6%) contained nuclei that, on average, were darker than or as dark as a PMN (extremely dark, ie, "India ink"). Fifty-one of 69 cases (74.0%) contained nuclei that, on average, were slightly brighter than a PMN (hyperchromatic). Ten of 69 cases (14.5%) contained nuclei that, on average, were much brighter than a PMN (hypochromatic). Within a single case, all cases showed heterogeneity with the hypochromatic cases showing the most dramatic effect. Digital image analysis reveals that there are large variations in chromasia between cases including a subset of cases with hypochromasia and another with extremely dark or "India ink" nuclei. There was much heterogeneity of chromasia seen within a single sample.

An Unusual Case of Urothelial Carcinoma of Bladder with Metastasis to Brain and Skeletal Muscles Evaluated on 18F-Fluorodeoxyglucose Positron Emission Tomography/Computed Tomography.

Urothelial or transitional cell carcinoma is the most prevalent variant of urinary bladder carcinoma worldwide occurring in both men and women. There are a very few cases described in the literature with metastasis to skeletal muscles and even fewer cases with metastatic site in the brain. We present a case of high-grade urothelial carcinoma with metastasis to the left temporal lobe as well as multiple inter- and intra-muscular deposits in the left thigh.

A Contemporary Clinicopathologic Analysis of Primary Urothelial Carcinoma of the Urethra Without Concurrent Renal Pelvic, Ureteral, or Bladder Carcinoma.

Primary urothelial carcinoma (UCa) of the urethra is relatively uncommon, and the underlying pathogenesis has not been well characterized, especially in the absence of concurrent UCa at other sites. A search for cases of primary UCa of the urethra was conducted. Patients with concurrent UCa of the renal pelvis, ureter, or bladder at the time of diagnosis of the primary tumor were excluded. Clinicopathologic and follow-up data were obtained. A total of 35 cases from 30 patients (27 male and 3 female) were included in the study. The mean patient age at the initial diagnosis was 71 years (range: 41-90 years). Cases were composed of high-grade UCa (26 of 35 = 74%), low-grade UCa (4 of 35 = 11%), and UCa in situ (5 of 35 = 14%). Invasion was present in 14 of 26 (54%) cases of high-grade UCa. Interestingly, 23 of 30 (77%) patients had a previous history of UCa including 7 (30%) cases with divergent differentiation or variant histology. Follow-up data were available in 23 patients with a mean duration of 26.7 months (range: 0.6-87 months). Eleven patients (31%) died of metastatic UCa. This is one of the largest studies to date of primary UCa of the urethra without concurrent UCa of the renal pelvis, ureter, or bladder. Previous history of UCa of the bladder, especially with divergent differentiation or variant histology is conceivably a key risk factor for developing subsequent primary UCa of the urethra. These findings are important for the development of surveillance protocols and therapeutic strategies.

Nuclear hypochromasia: Shedding light on the "lightness" of high-grade urothelial carcinoma.

The Paris system (TPS) has provided a standardized classification for reporting urinary cytology, specifically high-grade urothelial carcinoma (HGUC). While hyperchromasia is well described in HGUC, there exists little data on nuclear hypochromasia in HGUC. Our focus was to investigate the incidence of hypochromasia in HGUC and if it should become a criterion for HGUC. All cases of HGUC at our institution for a 3-year interval (2017-2019) using TPS were reviewed. Each case had a single ThinPrep slide and concurrent biopsy or resection specimen to confirm the diagnosis within 30 days. The presence of hypochromasia was evaluated, and cases with hypochromasia were stratified based on the tumor cell percentage. Cases with hypochromasia in 5% or greater of the tumor cells were considered "positively identified" for hypochromasia. We reviewed 117 cases of HGUC and identified nuclear hypochromasia in 12 cases (10.2%) within 5% or greater of the tumor cells. These 12 cases were further assessed based on if tumor cells showed hypochromasia in 5%-49% of the sample, or greater than 50% of the sample. Hypochromasia in 5%-49% of tumor cells was present in 8/117 cases (6.8%); whereas in 50% or greater samples 4/117 cases (3.4%) showed hypochromasia. No cases were identified where hypochromasia was noted in less than 5% of the tumor cells. TPS and use of hyperchromasia as a feature of HGUC is affirmed. However, hypochromasia, while not diagnostic in isolation, is present in a sub-set of patients with HGUC (10.2%) and should be considered as a variance noted in the nuclei of HGUC.

Diagnostic value of a comprehensive, urothelial carcinoma-specific next-generation sequencing panel in urine cytology and bladder tumor specimens.

Urine cytology can reliably diagnose high-grade urothelial carcinoma (HGUC) but not low-grade urothelial carcinoma (LGUC), and a more sensitive test is needed. Previously, a pilot study highlighted the possible diagnostic utility of next-generation sequencing (NGS) in identifying both LGUC and HGUC in urine cytology specimens. Twenty-eight urine ThinPrep cytology specimens and preceding or subsequent bladder tumor biopsy/resection specimens obtained within 3 months were included in the study (LGUC, n = 15; HGUC, n = 13). A customized, bladder-specific NGS panel was performed; it covered 69 frequently mutated or altered genes in urothelial carcinoma (UC) that were reported by The Cancer Genome Atlas and the Catalogue of Somatic Mutations in Cancer. The sequencing results were compared between the urine cytology specimens and the corresponding bladder tumor biopsies/resections. TP53 was the most frequently identified mutation in HGUC cases (11 of 13 [85%]). PIK3CA and KDM6A were the most frequently identified mutations in LGUC: they occurred in 7 of 15 cases (47%) and in 6 of 15 cases (40%), respectively. Additional frequent mutations identified in the panel included ARID1A (n = 5), EP300 (n = 4), LRP1B (n = 3), ERBB2 (n = 2), STAG2 (n = 2), FGFR3 (n = 3), MLL (n = 2), MLL3 (n = 2), CREBBP1 (n = 1), RB1 (n = 1), and FAT4 (n = 1). Overall, the concordance between the cytology and surgical specimens was 75%. The sensitivity and specificity for identifying mutations in urine cytology specimens were 84% and 100%, respectively. A bladder-specific NGS panel increases the sensitivity and specificity of urine cytology's diagnostic utility in both low- and high-grade tumors and may serve as a noninvasive surveillance method in the follow-up of patients with UC harboring known mutations.

Paraneoplastic Leukemoid Reaction after Primary Tumor Resection in Patients with Urothelial Carcinoma: A Report of 2 Cases

Leukemoid reaction (LR), which is defined as leukocytosis with a white blood cell (WBC) count above 50 000/µL, can be caused by various conditions, while paraneoplastic leukemoid reaction (PLR), a rare type of paraneoplastic syndrome, occurs in cases of solid tumors. Here we report 2 cases of high-grade urothelial carcinoma (HGUC) with PLR accompanied by rapid tumor progression after complete resection of the primary tumor. We reviewed the patient’s clinical history, histopathology, and the results of laboratory tests to rule out LR induced by non-tumor causes. In both cases, PLR appeared after primary tumor resection, and the patients died of disease at the peak of PLR at 6 and 8 weeks. Immunohistochemistry for granulocyte colony-stimulating factor and its receptor was performed on tumor tissues. Patients with HGUC and PLR are rare and have an extremely poor prognosis. The mechanism by which solid tumors are associated with PLR and rapid tumor progression after surgical resection of the primary tumor is incompletely understood and will be discussed here.

Developing a Machine Learning Algorithm for Identifying Abnormal Urothelial Cells: A Feasibility Study

Introduction: Urine cytology plays an important role in diagnosing urothelial carcinoma (UC). However, urine cytology interpretation is subjective and difficult. Morphogo (ALAB, Boston, MA, USA), equipped with automatic acquisition and scanning, optical focusing, and automatic classification with convolutional neural network has been developed for bone marrow aspirate smear analysis of hematopoietic diseases. The goal of this preliminary study was to determine the feasibility of developing a machine learning algorithm on Morphogo for identifying abnormal urothelial cells in urine cytology slides. Methods: Thirty-seven achieved abnormal urine cytology slides from cases with the diagnosis of atypical urothelial cells and above (suspicions or positive for UC) were obtained from 1 hospital. A pathologist (J.R.) reviewed the slides and manually selected and annotated representative cells to feed into Morphogo with following categories: benign (urothelial cells, squamous cells, degenerated cells, and inflammatory cells), atypical cells, and suspicious cells. Initial validation of the algorithm was performed on a subset of the original 37 cases. Urine samples from additional 12 unknown cases with various histological diagnoses (6 cases of high-grade urothelial carcinoma (HGUC), 1 case of low-grade urothelial carcinoma (LGUC), 1 case of prostate adenocarcinoma, 1 case of renal cell carcinoma, and 4 cases of non-neoplastic conditions) were collected from another hospital for initial blind testing. Results: A total of 1,910 benign and 1,978 abnormal (atypical and suspicious) cells from 37 slides were annotated for developing and training of the algorithm. This algorithm was validated on 27 slides that resulted in identification of at least 1 abnormal cell per slide, with a total of 200 abnormal cells, and an average of 7.4 cells per slide. Of the 12 unknown cases tested, the original cytology was positive for tumor cells in 2 HGUC samples. Morphogo was abnormal (atypical or suspicious) for 6 samples from patients with UC, including one with LGUC and one with prostate adenocarcinoma. Conclusion: Morphogo machine learning algorithm is capable of identifying abnormal urothelial cells. Further validation studies with a larger number of urine samples will be needed to determine if it can be used to assist the cytological diagnosis of UC.