Detection Of Urothelial Carcinoma Research Articles

Abstract 4878: Enhancing non-invasive detection of urothelial carcinoma through combined cytology and methylation profiling of urinary cell-free DNA

Abstract Detection of urothelial carcinoma (UC) faces a significant challenge in sensitivity due to the limitations of existing methodologies such as cytology. Identifying tumors through epigenetics pattern presents a promising advancement in diagnostic precision. This study aims to explore a non-invasive and more sensitive strategy by analyzing urinary cell-free DNA (ucfDNA) methylation profiles in conjunction with traditional urine cytology. The cohort for this study were collected from 20 patients with UC and 17 patients with benign or non-tumor lesions which histopathologic and cytopathologic results were reviewed by pathologist. Additional 16 cancer samples and 40 healthy donor samples were collected from independent research centers. We employed PredicineEPIC, a comprehensive whole-genome methylation assay, to investigate the DNA methylation characteristics in urine samples. The dataset was splited into 1:2 proportions for modeling and validation. The methylation signals were fed to three machine learning algorithms for determining the most effective classification model. Differentiating cancer and benign samples through urinary cytology has high specificity (1.0) but low sensitivity (0.3). Then we examined differentially methylated regions and implemented four most abnormally methylated regions as classify features. We found that the Gradient Boosting Machine (GBM) model achieved optimal performance in the validation set, with an AUC of 0.93, sensitivity of 0.81 (21/26), and specificity of 1.0 (36/36). The combination of methylation- and urine cytology-based models raised 2% prediction accuracy without reducing precision, suggesting that this assay could be applied to augment the cytology testing for UC detection. This study proposes a new and promising approach, leveraging cytology combined with epigenetic profiles of urinary cell-free DNA, for the precise diagnosis of urothelial carcinoma. Citation Format: Huan Zhao, Hang Dong, Haoran Tang, Zhihui Zhang, Na Wei, Jingjing Xu, Pan Du, Shidong Jia, Ting Xiao, Huiqin Guo. Enhancing non-invasive detection of urothelial carcinoma through combined cytology and methylation profiling of urinary cell-free DNA [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4878.

Diagnostic Immunocytostaining with Peroxisome Proliferator-Activated Receptor-Gamma in Urine Cytology Samples

Introduction: Urine cytology is a common method for detection of urothelial carcinoma (UC), however, is not high sensitivity. Improvement of the accuracy of cytodiagnosis using immunocytostaining as an auxiliary method is needed. This study aimed to determine the cytodiagnostic usefulness of peroxisome proliferator-activated receptor-gamma (PPAR-γ) immunocytostaining in urine cytology for the detection of UCs, particularly low-grade urothelial carcinomas (LGUC). Methods: PPAR-γ immunocytostaining was performed for 37 UC cases and 26 benign cases. Among the UC cases, 22 cases were of the papillary proliferation type, not including the mixed type comprising both papillary and flat growth. Fifteen LGUC cases of all papillary proliferation types were included. For comparison, the same samples were also immunocytostained for p53 and Ki-67. Results: Of the UC cases, 25 of 37 were positive for PPAR-γ, while 24 of the 26 benign cases were PPAR-γ-negative. Regardless of histological grading, 13 of the 22 UC cases with papillary proliferation were PPAR-γ-positive. In particular, PPAR-γ immunocytostaining showed higher sensitivity for LGUC cases than that of the other biomarkers. Regarding LGUC specifically, 4 of 10 cases not identified by primary cytology were detected by PPAR-γ immunocytostaining. Conclusion: PPAR-γ immunocytostaining enhances the accuracy of urine cytodiagnosis. Furthermore, PPAR-γ is a more useful immunobiomarker in urine cytology than p53 and Ki-67, the commonly used immunobiomarkers for malignant cell detection.

MCM4 expression is associated with high-grade histology, tumor progression and poor prognosis in urothelial carcinoma

BackgroundWe previously reported Minichromosome maintenance 4 (MCM4) overexpression in gastric cancer. However, the clinicopathological significance of MCM4 in urothelial carcinoma (UC) has not been investigated. To clarify the clinicopathological significance of MCM4 in UC, we investigated MCM4 expression with immunohistochemistry (IHC).MethodsWe analyzed the expression and distribution of MCM4 in 124 upper tract urothelial carcinoma (UTUC) samples by IHC. Additionally, using 108 urine samples, we analyzed MCM4 Immunocytochemistry (ICC) expression in urine cytology.ResultsIn normal urothelium, MCM4 expression was weak or absent. Meanwhile, the strong nuclear expression of MCM4 was observed in UTUC tissues, and it was detected in 77 (62%) of a total of 124 UTUC cases. MCM4-positive UTUC cases were associated with nodular/flat morphology, high grade, high T stage, and poor prognosis. Moreover, MCM4 expression was significantly higher in the invasive front than in the tumor surface. Similar results were also obtained in TCGA bladder cancer cohort. Additionally, MCM4 expression was associated with high expression of Ki-67, HER2, EGFR, and p53 in UTUC. Among representative cancer-related molecules, MCM4 had an independent predictive value for progression-free survival and high-grade UC. ICC for MCM4 was also performed on urine cytology slides and showed that the nuclear expression of MCM4 was more frequently found in UC cells than in non-neoplastic cells. The diagnostic accuracy of urine cytology was improved by combining MCM4 immunostaining with cytology.ConclusionThese results suggest that MCM4 might be a useful predictive biomarker for high-grade histology, tumor progression and poor prognosis in UC. Moreover, ICC for MCM4 might be helpful for UC detection as additional markers in the cytomorphology-based diagnosis.

Dual immunocytochemical staining of annexin A10 and p53 in low-grade and papillary urothelial carcinoma contributes to improvement of diagnostic accuracy in urine cytology.

Urothelial carcinoma (UC) is a common type of human cancer and, although urine cytology is a useful method for identifying high-grade UC (HGUC), its ability to diagnose low-grade UC (LGUC) is limited. The authors previously reported that annexin A10 (ANXA10) expression was strongly linked to both papillary and early stage LGUC and was inversely correlated with p53 expression in upper tract UC (UTUC) and bladder UC. However, it remains largely unknown whether ANXA10 is useful as a diagnostic marker for urine cytology. In this study, the authors used 104 biopsy and 314 urine cytology samples to investigate the efficacy of ANXA10 and p53 expression by immunohistochemistry and immunocytochemistry. In immunohistochemistry analysis, expression levels of ANXA10 and p53 were either weak or absent in noncancerous tissues, whereas ANXA10 overexpression was observed patients with LGUC, and strong expression of p53 was identified in patients with HGUC. In immunocytochemistry analysis, sensitivity was not good for the detection of UC, especially UTUC, by cytology alone, but it was improved by combining cytology with ANXA10 and p53 to detect both bladder UC and UTUC. Receiver operating characteristic curve analysis also confirmed the diagnostic superiority of cytology combining ANXA10 and p53 for the detection of all UCs, including both HGUC and LGUC (area under the curve, 0.84). To the authors' knowledge, this is the first report that the combination of ANXA10 and p53 has potential application as a diagnostic immunomarker for improving the diagnostic accuracy of urine cytology.

UriFind assay and association with whole course management of urothelial carcinoma in a large multicentric, prospective China cohort.

e16588 Background: The gold standard to diagnose and monitor urothelial carcinoma is cystoscopy, which is invasive and painful. The currently available non-invasive methods to detect urothelial carcinoma are either of unsatisfying sensitivity or limited specificity in China. Previously, a DNA methylation assay, termed UriFind, was developed, and showed a compelling sensitivity and specificity in a retrospective cohort. This study aimed to further validate its performance in a large multicentric, prospective China cohort. Methods: Patients involved in the whole course of UC management were recruited in this cohort. The cohort contained 931 patients diagnosed with urothelial carcinoma (UC), and 994 controls with benign diseases and 78 healthy participants from 12 hospitals across China. Urine samples were collected before the therapy, and analyzed with quantitative PCR of ONECUT2 and VIM methylation (UriFind), FISH and urinary cytology. Sensitivity, specificity, accuracy, PPV and NPV of these assays were evaluated as compared to the diagnostic gold standard of cystoscopy, histology results or clinical diagnosis. Results: The UC group of the cohort consisted of 683 non-muscle invasive UC (73.4%), 203 muscle invasive UC (21.8%) and 45 unavailable stage (4.8%). The patients of low, high-grade UC and unavailable grade were 244(26.2%), 651(69.9%) and 36 (3.9%), respectively. Overall sensitivity, specificity, accuracy, PPV and NPV of UriFind was 82.3%, 91.2%, 87.3%, 88.1% and 86.7%, respectively. The specificity of healthy participants was 100%. The overall sensitivity and accuracy of UriFind were significantly higher than urine cytology (40% and 55.7%) and FISH (68.2% and 73.6%), while the specificity was similar to cytology (93.9%) and FISH (87.3%). Importantly, UriFind achieved a great improvement of sensitivity in whole course management, including low grade tumor (56.1%), small and single tumor (63.3%), residual tumor after therapy (71.4%) and recurrent tumor (84.3%). Conclusions: The performance of UriFind is well validated with significantly higher sensitivity and accuracy than urine cytology and FISH in UC detection in this cohort with 2003 cases. This assay facilitates whole course management of urothelial carcinoma, which may reduce the burden of cystoscopy and unnecessary second TURBT. Clinical trial information: NCT04314245 .

Tumor cell-based glycolytic metabolism and single-cell sequencing of urinary exfoliated cells for the diagnosis and molecular profiling of urothelial carcinoma

Objective: To explore the diagnostic values of HK2 testing and single-cell sequencing in the urothelial carcinoma (UC). Methods: The qualified urine specimens of 265 suspected UC patients or postoperative patients from the Cancer Hospital of Chinese Academy of Medical Sciences, Beijing, China were collected. Both exfoliative cytology and HK2 testing were performed on clinically suspected UC or postoperative patients. The performance of diagnostic cytology and HK2, including consistency, sensitivity, specificity, positive predictive value and negative predictive value, was evaluated based on histopathological, clinical and imaging diagnosis. Isolated HK2 metabolically abnormal cells were subject to single-cell sequencing to verify the reliability of HK2 detection performance and to explore the molecular characteristics of UC. Results: The concordance rate of HK2 testing and cytology for detecting UC was 90.3% (102/113, Kappa=0.604). Compared with cytology, the sensitivity of HK2 was significantly higher (85.2% versus 75.6%, P=0.024). The detection sensitivity of combined HK2 testing and cytology was increased to 91.1%. HK2 testing was significantly more sensitive than cytology for diagnosing UC in the upper urinary tract (81.8% versus 65.5%, P=0.022). It was also more sensitive than cytology for diagnosing early-stage UC (82.6% versus 69.5%, P=0.375) and low-grade UC (69.6% versus 47.8%, P=0.125). Single-cell sequencing of the ten patients, whose samples were positive for HK2, demonstrated highly concordant copy number variations (CNVs) in tumor cells from the same UC patient, with heterogeneity in CNV profiles among different patients. Deletion of chromosome 8p was found in 3 of the 4 urine samples of renal pelvis UC. The 2 patients with benign lesions had no CNVs in all sequenced cells. Conclusions: The test for abnormal urinary glycolytic HK2 metabolism can assist urine cytology to improve the sensitivity of UC diagnosis, and it provides a novel and reliable approach for early detection of upper urinary tract UC and lower grade UC. Meanwhile, this study has preliminarily revealed the feasibility of single-cell sequencing in urinary samples, which is expected to improve the diagnostic specificity of HK2 testing.

MP63-18 UriFIND ASSAY FACILITATES WHOLE COURSE MANAGEMENT OF UROTHELIAL CARCINOMA IN A LARGE MULTICENTRIC, PROSPECTIVE CHINA COHORT

MP63-18 UriFIND ASSAY FACILITATES WHOLE COURSE MANAGEMENT OF UROTHELIAL CARCINOMA IN A LARGE MULTICENTRIC, PROSPECTIVE CHINA COHORT

MP22-19 URINARY COMPREHENSIVE GENOMIC PROFILING PREDICTS UROTHELIAL CANCER UP TO 12 YEARS AHEAD OF CLINICAL DIAGNOSIS. AN EXPANDED ANALYSIS OF THE GOLESTAN COHORT STUDY

MP22-19 URINARY COMPREHENSIVE GENOMIC PROFILING PREDICTS UROTHELIAL CANCER UP TO 12 YEARS AHEAD OF CLINICAL DIAGNOSIS. AN EXPANDED ANALYSIS OF THE GOLESTAN COHORT STUDY

CDCP1 (CUB domain containing protein 1) is a potential urine-based biomarker in the diagnosis of low-grade urothelial carcinoma.

Urine-based cytology is non-invasive and widely used for clinical diagnosis of urothelial carcinoma (UC), but its sensitivity is less than 40% for low-grade UC detection. As such, there is a need for new diagnostic and prognostic biomarkers of UC. CUB domain containing protein 1 (CDCP1) is a type I transmembrane glycoprotein highly expressed in various cancers. Using tissue array analysis, we demonstrated that CDCP1 expression in UC patients (n = 133), especially in those with low-grade UC, was significantly higher than in 16 normal persons. In addition, CDCP1 expression in urinary UC cells could also be detected by using immunocytochemistry method (n = 11). Furthermore, in 5637-CD cells, overexpression of CDCP1 affected the expression of epithelial mesenchymal transition-related markers and increased matrix metalloproteinase 2 expression and migration ability. Conversely, the knockdown of CDCP1 in T24 cells had the opposite effects. Using specific inhibitors, we demonstrated the involvement of c-Src/PKCδ signaling in the CDCP1-regulated migration of UC. In conclusion, our data suggest that CDCP1 contributes to the malignant progression of UC and may have the potential as a urine-based biomarker for detecting low-grade UC. However, a cohort study needs to be conducted.

Survey of keratin 17 (K17) expression in different urothelial malignancies.

458 Background: The current non-invasive urine tests tend to display low sensitivity for the detection of urothelial cancers, especially in low-grade cancer. Recent reports suggest that the URO17 test based on the Keratin 17 (K17) biomarker displays high sensitivity and specificity in the detection of urothelial carcinoma in both low-grade and high-grade cancers and show promise as an effective non-invasive test for bladder cancer. The current study aimed to evaluate K17 expression in different histological categories of urological diseases. Methods: Ninety-five urine specimens were collected from 2020-2021 that represented 19 different histological categories of urothelial diseases. These samples were prepared onto slides and tested with the URO17 Bladder Cancer test (KDx Diagnostics Inc.) on DAKO Link48 autostainer using the manufacturer’s recommended procedures. The stained samples were evaluated by a cytopathologist and scored using five K17-positive urothelial cells as a cut-off. Results: K17 was positive in 20/20 (100%) of the urothelial carcinoma, and 23/27 (85%) of Non-invasive papilloma and urothelial carcinomas. K17 was positive in 100% of carcinoma in situ, papillary urothelial carcinoma, sarcoma, and small cell carcinoma, papillary urothelial carcinoma, urothelial papilloma, inverted papilloma, renal-pelvis urothelial carcinoma, and urothelial carcinoma with squamous differentiation. K17 was also expressed in many cystitis, inflammatory myofibroblastic tumors, and PUNLMP, but not in prostate carcinoma. In 19/22 cases (86%) K17 was mostly positive in urine samples from patients with prior history of bladder cancer, but were histologically determined to be benign. The overall sensitivity of K17 in the cancer group was 88.5% (54/61 positive) with K17 being expressed in both high-grade and low-grade cancers. Of the seven false-negative samples, five were low-grade, one was high-grade, and one was from the inflammatory myofibroblastic tumor. Conclusions: Keratin 17 is highly expressed in almost all cases of urothelial carcinoma including those of papillary and squamous characteristics, confirming the high sensitivity of K17 in detecting all categories of urothelial carcinoma. Keratin 17 was also observed in many benign and premalignant conditions such as cystitis, PUNLMP, and metaplasia suggesting the K17 expression can result from a trauma to urothelium that could lead to activation of K17 in regenerating urothelial cells. Additional studies are ongoing to confirm and expand the utility of the K17 biomarker in bladder cancer.

Independent verification of circulating miRNA as diagnostic biomarkers for urothelial carcinoma.

Urothelial carcinoma (UC) is an umbrella term for bladder cancers (BCa) and upper‐tract urothelial carcinoma (UTUC), with BCa and UTUC sometimes detected concomitantly. The methods of detection for UC are often inaccurate or highly invasive, and, therefore, are thought to be unsatisfactory. Previously, we reported seven serum miRNAs as diagnostic markers for BCa. Here, we re‐evaluated potential diagnostic miRNAs in different institutions. We prospectively analyzed serum samples obtained from 126 UC patients (BCa: 106 samples; UTUC: 14 samples; UTUC with BCa: six samples) and 50 noncancer controls by microarray analysis. We randomly assigned these samples into a training or a validation set. Biomarker candidate miRNAs were selected based on cross‐validation scores in the training set of samples, with diagnostic power confirmed in the validation set. Among the diagnostic miRNAs identified in this way, miR‐1343‐5p and miR‐6087 had been identified as potential diagnostic miRNAs in our previous study. In addition, we evaluated the association between the serum levels of identified miRNAs and the presence of UC risk conditions. The expression levels of several miRNAs correlate with the risk factors in participants without UC, which may be explained by the presence of a microscopic tumor or a precancerous lesion. In conclusion, we identified two robust miRNA diagnostic markers for UC detection. Further functional analysis is required to elucidate the mechanism by which alterations in the expression of these miRNAs occur.

Management of secondary Paget's disease of the vulva associated with transitional cell carcinoma.

Secondary extramammary Paget's disease (EMPD) related to urothelial carcinoma is rare, with some cases presenting synchronously with either a primary neoplasm or recurrence of a neoplasm and other cases presenting up to 13years prior to the detection of urothelial carcinoma. In this report, we will review the presentation, diagnosis, pathophysiology, management, and literature review of cases of secondary EPMD associated with urothelial carcinoma. We reviewed the English literature for all cases of secondary EMPD presenting synchronously with or in patients with a history of urothelial carcinoma, as well as treatment data for secondary vulvar Paget's. We identified 16 case reports and case series with a total of 20 cases of vulvar EMPD associated with urothelial carcinoma. Twelve cases presented asynchronously and 8 had EMPD preceding the diagnosis of the underlying neoplasm. There is a paucity in the literature regarding management and surgical resection is a common treatment strategy; however, nonsurgical interventions may also be effective. There is a paucity in the literature regarding management of secondary EPMD of urothelial origin, but consideration of radiation and systemic chemotherapy may be a reasonable treatment approach.

Comprehensive genomic profiling of urine DNA for urothelial carcinoma detection and risk prediction.

450 Background: Clinical diagnosis and risk stratification of patients with urothelial carcinoma (UC) remains a challenge, with high rates of recurrence and disease progression following treatment. Urinary comprehensive genomic profiling (uCGP) has significant potential to aid in both diagnosis and prognostication of non-muscle-invasive and muscle-invasive disease. Methods: uCGP was performed on urine specimens collected at 9 centers across the US from 577 subjects prior to cystoscopy. 152 subjects were UC tumor positive (de novo and recurrence), 191 had a history of UC but negative by surveillance cystoscopy at time of collection, and 234 were urology control subjects undergoing cystoscopy without evidence of UC. Urine DNA was sequenced and comprehensively profiled across 60 genes for 6 classes of mutations using the CLIA-validated UroAmplitude test. Disease detection and molecular grade (high grade vs. low grade) algorithms were trained (n=345) and validated (n=232) in independent cohorts. Results: Among UC tumor positives, grade distribution was 53% high grade, 41% low grade, and 6% unknown. Stage distribution was Tis (5%), Ta (57%), T1 (16%), ≥T2 (15%), Tx (7%). 99% of tumor positive patients had one or more mutation identified. Interestingly, 69% of UC surveillance negative and 49% of urology controls also had at least one high impact mutation. The prevalence of mutations among controls necessitates machine learning algorithms to classify disease status. In validation, de novo tumor diagnosis demonstrated sensitivity of 93.8% and specificity of 89.4% and a NPV of 98.8% in urology controls. Recurrent tumors were detected with a PPV of 73.5%, sensitivity of 62.5% and specificity of 89.0% in patients with a history of UC. Molecular grading predicted high-grade with a PPV of 90.9% and a specificity of 96.7% compared to pathology. Urinary TP53 mutations were enriched in ≥T2 tumors relative to Ta (OR=14.8 [95%CI 4.6-47.5], P=0.00001). Copy number alterations were also associated with increased risk of muscle invasion, metastasis, and enriched for CIS relative to Ta tumors (≥T2: OR=6.4 [95%CI 1.8-22.9], P=0.019; CIS: OR=10.5 [95%CI 1.9-58.9], P=0.04). Conclusions: We developed and validated a uCGP test that provides robust noninvasive detection of UC across a diverse group of patients and clinical contexts, including non-muscle-invasive and muscle-invasive UC. Mutations with actionable or prognostic value are found in most subjects. These data suggest that uCGP classifies tumor presence with better performance than traditional urinary biomarkers. Importantly, uCGP identifies genomic markers of muscle invasion, metastasis, and CIS. With longer term follow-up, uCGP mutational profiles may reveal important prognostic information regarding risk of disease recurrence and progression. Additional studies are underway to further support the generalizability of these findings.

A0241 - Urine cellular DNA point mutation and methylation as potential biomarkers for early detection of urothelial carcinoma

A0241 - Urine cellular DNA point mutation and methylation as potential biomarkers for early detection of urothelial carcinoma

MP48-01 THE N-GLYCAN SIGNATURE OF SERUM IMMUNOGLOBULINS IS A DIAGNOSTIC BIOMARKER OF UROTHELIAL CARCINOMAS

You have accessJournal of UrologyBladder Cancer: Upper Tract Transitional Cell Carcinoma II (MP48)1 Sep 2021MP48-01 THE N-GLYCAN SIGNATURE OF SERUM IMMUNOGLOBULINS IS A DIAGNOSTIC BIOMARKER OF UROTHELIAL CARCINOMAS Hirotake Kodama, Tohru Yoneyama, Takuma Narita, Yuta Kojima, Hiromichi Iwamura, Teppei Okamoto, Hayato Yamamoto, Shingo Hatakeyama, Takahiro Yoneyama, Yasuhiro Hashimoto, and Chikara Ohyama Hirotake KodamaHirotake Kodama More articles by this author , Tohru YoneyamaTohru Yoneyama More articles by this author , Takuma NaritaTakuma Narita More articles by this author , Yuta KojimaYuta Kojima More articles by this author , Hiromichi IwamuraHiromichi Iwamura More articles by this author , Teppei OkamotoTeppei Okamoto More articles by this author , Hayato YamamotoHayato Yamamoto More articles by this author , Shingo HatakeyamaShingo Hatakeyama More articles by this author , Takahiro YoneyamaTakahiro Yoneyama More articles by this author , Yasuhiro HashimotoYasuhiro Hashimoto More articles by this author , and Chikara OhyamaChikara Ohyama More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002074.01AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Discriminating between urothelial carcinoma (UC) including bladder cancer (BCa) and upper urinary tract UC (UTUC) is often challenging. METHODS: We evaluated the diagnostic performance of BCa and UTUC discrimination scores that we developed based on N-glycosylation signatures of immunoglobulins (Igs) for the detection of UC including BCa and UTUC. The N-glycosylation signatures of Igs from serum samples of in 405 patients with UC (176 with BCa, 104 with UTUC), 21 with urinary tract infection, and 104 healthy volunteers were measured retrospectively using automated capillary-electrophoresis-based N-glycomics analysis. The UTUC or BCa discriminating scores were established by discriminant analysis using the N-glycan signatures of Igs. Diagnostic performance was evaluated using the area under receiver operating characteristics curve (AUC) and decision curve analyses (DCA). RESULTS: The BCa score for discriminating BCa (AUC: 0.979) and UTUC score for discriminating UTUC (AUC: 0.831) provided significantly better clinical performance than urine cytology negative or gross hematuria negative or clinical T1 cases. The DCA analysis revealed that adding the BCa and UTUC scores to gross hematuria status avoided the need for more intervention without missing UC (risk threshold: 8%–95%). We found that a UC diagnostic nomogram based on the combination of gross hematuria, UTUC score and BCa score could detect UC with an AUC of 0.939, positive predictive value of 91.8%, and negative predictive value of 75.7%. CONCLUSIONS: The UC score based on N-glycosylation signatures of Igs is a promising approach for the diagnosis of UC. Source of Funding: none © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e862-e863 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Hirotake Kodama More articles by this author Tohru Yoneyama More articles by this author Takuma Narita More articles by this author Yuta Kojima More articles by this author Hiromichi Iwamura More articles by this author Teppei Okamoto More articles by this author Hayato Yamamoto More articles by this author Shingo Hatakeyama More articles by this author Takahiro Yoneyama More articles by this author Yasuhiro Hashimoto More articles by this author Chikara Ohyama More articles by this author Expand All Advertisement Loading ...

Asymptotic modeling of electrochemical signaling: Testing Zn in urine for non-invasive bladder cancer diagnosis

A theoretical model on chemical signaling for diagnosis based on the combination of signals for marker and inert and/or interfering metabolites is described. The model yields asymptotic relationships between the intensities of the signals representative of marker and accompanying metabolites compensating concentration fluctuations. The model fits well with voltammetric features associated to the oxidation of different urine metabolites and Zn(II) reduction in the same urine samples after alkaline digestion. As a result, a non-invasive electrochemical detection of urothelial carcinoma (bladder cancer, BC) is reported. Different diagnostic criteria are described all displaying high sensitivity, specificity and positive/negative predictive values. The described methodology was tested by means of an analytical trial with 46 patients with histological and pathologically confirmed BC, 30 healthy controls and 26 patients diagnosed of other urinary pathologies,

Toward urinary cell-free DNA-based treatment of urothelial carcinoma: a narrative review.

Liquid biopsy technique targeting urinary cell-free DNA (cfDNA) is getting a lot of attention to overcome limitations of the present treatment strategy for urothelial carcinoma, including urothelial bladder carcinoma (UBC) and upper tract urothelial carcinoma (UTUC). Analysis of tumor-derived DNA in urine focusing either on genomic or epigenomic alterations, holds great potential as a noninvasive method for the detection of urothelial carcinoma with high accuracy. It is also predictive of prognosis and response to drugs, and reveals the underlying characteristics of different stages of urothelial carcinoma. Although cfDNA methylation analyses based on a combination of several methylation profiles have demonstrated high sensitivity for UBC diagnosis, there have been few reports involving epigenomic studies of urinary cfDNA. In mutational analyses, frequent gene mutations (TERT promoter, TP53, FGFR3, PIK3CA, RAS, etc.) have been detected in urine supernatant by using remarkable technological innovations such as next-generation sequencing and droplet digital PCR. These methods allow highly sensitive detection of rare mutation alleles while minimizing artifacts. In this review, we summarize the current insights into the clinical applications of urinary cfDNA from patients with urothelial carcinoma. Although it is necessary to conduct prospective multi-institutional clinical trials, noninvasive urine biopsy is expected to play an important role in the realization of precision medicine in patients with urothelial carcinoma in the near future.

Biomarkers for Precision Urothelial Carcinoma Diagnosis: Current Approaches and the Application of Single-Cell Technologies.

Simple SummaryUrothelial carcinoma (UC) is the most frequently diagnosed cancer of the urinary tract and is ranked the sixth most diagnosed cancer in men worldwide. About 70–75% of newly diagnosed UCs are non-invasive or low grade. Different tests such as urine cytology and cystoscopy are used to detect UC. If abnormal tissue is found during cystoscopy, then a biopsy will be performed. Cytology has low sensitivity for low-grade cancer while cystoscopy is invasive and costly. Detecting UC early improves the chances of treatment success. Therefore, many researchers have painstakingly identified urine biological markers for non-invasive UC diagnosis. In this review, we summarize some of the latest and most promising biological markers (including FDA-approved and investigational markers). We also discuss some new technologies that can aid research efforts in biological marker discovery for early UC detection.Urothelial carcinoma (UC) is the most frequent malignancy of the urinary system and is ranked the sixth most diagnosed cancer in men worldwide. Around 70–75% of newly diagnosed UC manifests as the non-muscle invasive bladder cancer (NMIBC) subtype, which can be treated by a transurethral resection of the tumor. However, patients require life-long monitoring due to its high rate of recurrence. The current gold standard for UC diagnosis, prognosis, and disease surveillance relies on a combination of cytology and cystoscopy, which is invasive, costly, and associated with comorbidities. Hence, there is considerable interest in the development of highly specific and sensitive urinary biomarkers for the non-invasive early detection of UC. In this review, we assess the performance of current diagnostic assays for UC and highlight some of the most promising biomarkers investigated to date. We also highlight some of the recent advances in single-cell technologies that may offer a paradigm shift in the field of UC biomarker discovery and precision diagnostics.

Noninvasive Detection of Urothelial Carcinoma by Cost-effective Low-coverage Whole-genome Sequencing from Urine-Exfoliated Cell DNA

Urothelial carcinoma is a malignant cancer with frequent chromosomal aberrations. Here, we investigated the application of a cost-effective, low-coverage whole-genome sequencing technology in detecting all chromosomal aberrations. Patients with urothelial carcinomas and nontumor controls were prospectively recruited in clinical trial NCT03998371. Urine-exfoliated cell DNA was analyzed by Illumina HiSeq XTen, followed by genotyping with a customized bioinformatics workflow named Urine Exfoliated Cells Copy Number Aberration Detector (UroCAD). In the discovery phase, urine samples from 126 patients with urothelial carcinomas and 64 nontumor disease samples were analyzed. Frequent chromosome copy-number changes were found in patients with tumor as compared with nontumor controls. A novel diagnosis model, UroCAD, was built by incorporating all the autosomal chromosomal changes. The model reached performance of AUC = 0.92 (95% confidence interval, 89.4%-97.3%). At the optimal cutoff, |Z| ≥ 3.21, the sensitivity, specificity, and accuracy were 82.5%, 96.9%, and 89.0%, respectively. The prediction positivity was found correlated with tumor grade (P = 0.01). In the external validation cohort of 95 participants, the UroCAD assay identified urothelial carcinomas with an overall sensitivity of 80.4%, specificity of 94.9%, and AUC of 0.91. Meanwhile, UroCAD assay outperformed cytology tests with significantly improved sensitivity (80.4% vs. 33.9%; P < 0.001) and comparable specificity (94.9% vs. 100%; P = 0.49). UroCAD could be a robust urothelial carcinoma diagnostic method with improved sensitivity and similar specificity as compared with cytology tests. It may be used as a noninvasive approach for diagnosis and recurrence surveillance in urothelial carcinoma prior to the use of cystoscopy, which would largely reduce the burden on patients.

Cytologically targeted next-generation sequencing: a synergy for diagnosing urothelial carcinoma

Cytology and cystoscopy are used to detect urothelial carcinoma (UC), but together they still fail to detect some UC cases and are not suitable for screening asymptomatic individuals. Mutations are present in more than 98% of UC, mutations have therapeutic significance, and they can be detected by next generation sequencing (NGS) in urine samples. We review the role of NGS in UC detection. Comprehensive literature review on UC genetics, economics of NGS, and previous reports of UC detection by NGS. The raw costs of NGS have decreased to about 14,000 base pairs per penny, making it appear economically feasible to use NGS widely. Reported NGS assays fall short of predicted sensitivity. Decreased sensitivity is attributed to a low frequency of mutant alleles in many urine samples. Attempts to increase the percentage of mutant alleles, by using cell-free urinary DNA, or by using cell sorting and microfluidics, have been unsuccessful or remain unproven. However, cytologic examination can immediately enable NGS: Urine cytologies with sufficient proportions of abnormal cells could be directly triaged to NGS with high sensitivity for UC detection. For cases with a low proportion of abnormal cells, cytologically targeted microdissection of cells for NGS should maintain sensitivity and decrease sequencing costs. Cytologically targeted urothelial cells for NGS could allow a screening test for low grade UC. Cytology is immediately poised to allow NGS to improve the diagnosis of UC, allowing NGS to be an ancillary test for atypical cytologies, and potentially allowing a screening test for low-grade UC.