You have accessJournal of UrologyPenile & Testicular Cancer II (MP61)1 May 2024MP61-08 TROP-2—A PROMISING NEW THERAPEUTIC TARGET IN PENILE SQUAMOUS CELL CARCINOMA Richard Weiten, Carolina Kessler, Christina Meisl, Hanna Elisa Spohn, Enno Storz, David Pfister, Tim Nestler, Yuri Tolkach, Friederike Linden, Ralph Wirtz, Melanie von Brandenstein, Philipp Krausewitz, and Axel Heidenreich Richard WeitenRichard Weiten , Carolina KesslerCarolina Kessler , Christina MeislChristina Meisl , Hanna Elisa SpohnHanna Elisa Spohn , Enno StorzEnno Storz , David PfisterDavid Pfister , Tim NestlerTim Nestler , Yuri TolkachYuri Tolkach , Friederike LindenFriederike Linden , Ralph WirtzRalph Wirtz , Melanie von BrandensteinMelanie von Brandenstein , Philipp KrausewitzPhilipp Krausewitz , and Axel HeidenreichAxel Heidenreich View All Author Informationhttps://doi.org/10.1097/01.JU.0001009536.58867.87.08AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Penile squamous cell carcinoma (PSCC) is an infrequent yet progressively prevalent malignancy. Despite ongoing advancements in patients with the diagnosis of locally advanced or metastasized PSCC especially systemic treatment options are limited. Therefore, there is an urgent need for efficient salvage therapies. Recently in other solid tumor entities, e.g., breast and urothelial cancer, antibody-drug-conjugates (ADCs) demonstrated significant clinical efficacy. Our aim was to ascertain the potential utility of ADCs targeting TROP-2 in both localized PSCC (PRIM) and patient-matched distant metastases (MET). METHODS: Immunohistochemical analysis of TROP-2 expression was conducted in PRIM (n=40) and their corresponding METs (n=9, including lymph node and lung metastases). We assessed overall survival (OS) by stratifying patients into subgroups based on TROP-2 expression: weak (H-score 15–99) versus moderate/strong (H-score 100–300). Additionally, we explored the relationship between TROP-2 and HPV-dependency. Furthermore, TROP-2 mRNA expression levels were quantified via qRT-PCR, and an enzyme-linked immunosorbent assay was employed to measure TROP-2 in serum samples derived from PRIM (n=40) and a healthy control group (n=10). RESULTS: We found a statistically significant increase in TROP-2 mRNA expression levels in individuals afflicted with PSCC compared to healthy controls (p<0.0001). Moreover, a notable elevation in TROP-2 protein expression was observed in the serum specimens obtained from PSCC patients (p<0.01). The majority of patients exhibited moderate to strong TROP-2 expression, indicative of a robust median H-score of 280 (interquartile range: 210–300). Subgroup analysis revealed a correlation between TROP-2 expression and HPV status (p<0.01), demonstrating discernible associations with more favorable clinical outcomes, as evidenced by the log-rank (Mantel-Cox) test (p<0.05). CONCLUSIONS: TROP-2 protein expression in PSCC holds prognostic significance. Furthermore, the elevated TROP-2 levels in both PRIM and PSCC metastases suggest that the TROP-2-targeted ADC, Sacituzumab govitecan or Datopotamab deruxtecan, may emerge as a promising novel therapeutic intervention for these patients. Download PPT Source of Funding: N/A © 2024 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 211Issue 5SMay 2024Page: e1013 Advertisement Copyright & Permissions© 2024 by American Urological Association Education and Research, Inc.Metrics Author Information Richard Weiten More articles by this author Carolina Kessler More articles by this author Christina Meisl More articles by this author Hanna Elisa Spohn More articles by this author Enno Storz More articles by this author David Pfister More articles by this author Tim Nestler More articles by this author Yuri Tolkach More articles by this author Friederike Linden More articles by this author Ralph Wirtz More articles by this author Melanie von Brandenstein More articles by this author Philipp Krausewitz More articles by this author Axel Heidenreich More articles by this author Expand All Advertisement PDF downloadLoading ...

ObjectiveTo evaluate the potential utility of antibody‐drug conjugates targeting trophoblast cell surface antigen‐2 (TROP‐2) in patients with primary penile squamous cell carcinoma (PSCC), patients with recurrence (REC cohort), and patient‐matched distant metastases (MET cohort), and to assess the potential use of TROP‐2 as a predictive non‐invasive biomarker in PSCC.MethodsA cohort comprising a PRIM (n = 37), REC (n = 5) and MET subcohort (n = 7), with MET including lymph node and lung metastases, was analysed using quantitative real‐time PCR, ELISA and immunohistochemical staining with evaluation of H‐score.ResultsTROP‐2 mRNA and serum protein levels were significantly increased in primary and recurrent PSCC compared to cancer‐free controls (both P < 0.001). Immunohistochemical analysis revealed that most of the PRIM cohort (n = 34/37, median H‐score 260, interquartile range [IQR] 210–300), as well as all patients in the REC (median [IQR] H‐score 200 [165–290]) and MET cohorts (median [IQR] H‐score 280 [260–300]) exhibited moderate to strong membranous TROP‐2 expression. Additionally, The H‐score (membranous TROP‐2 expression) was positively correlated with TROP‐2 mRNA (ρ = 0.69, P < 0.0001, R2 = 0.70) and protein levels (ρ = 0.86, P < 0.0001, R2 = 0.59), indicating its potential as a non‐invasive biomarker in PSCC.ConclusionIn summary, our results support further studies on TROP‐2 as a diagnostic and therapeutic target in primary, recurrent and metastatic PSCC.

You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology I (MP15)1 May 2024MP15-01 NOVEL URINE-BASED BIOMARKER TO DISTINGUISH MUSCLE-INVASIVE FROM NON-MUSCLE INVASIVE BLADDER CANCER–A MULTICENTRIC APPROACH Barbara Koeditz, Lucas Kastner, Constantin Rieger, Sofia Semko, Maksyn Pikul, Enno Storz, Throsten Ecke, Friederike Linden, Ralph Wirtz, Richard Weiten, Melanie von Brandenstein, and Axel Heidenreich Barbara KoeditzBarbara Koeditz , Lucas KastnerLucas Kastner , Constantin RiegerConstantin Rieger , Sofia SemkoSofia Semko , Maksyn PikulMaksyn Pikul , Enno StorzEnno Storz , Throsten EckeThrosten Ecke , Friederike LindenFriederike Linden , Ralph WirtzRalph Wirtz , Richard WeitenRichard Weiten , Melanie von BrandensteinMelanie von Brandenstein , and Axel HeidenreichAxel Heidenreich View All Author Informationhttps://doi.org/10.1097/01.JU.0001009500.87761.bf.01AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: The truncated variant of mitogen-activated protein kinase p38 (MAPK p38), Mxi-2 is associated with the occurrence and progression of renal cell carcinoma and prostate cancer (PCa). Furthermore, Vimentin3 (Vim3)represent the truncated variant of vimentin, a key structural protein. The expression of Vim3 has already been linked to the presence of PCa biomarkers in serum, which facilitates the detection of renal cell carcinoma. Ouraim was to ascertain the potential utility of Mxi-2 and Vim3 as potential biomarkers for stratification between non-muscle invasive (NMBIC) and muscle-invasive bladder cancer (MBIC). METHODS: Immunohistochemical and immunofluorescence analysis of Mxi-2 and Vim3 were determined in localized urothelial carcinoma (n=30). Furthermore, corresponding urine samples of patient with localized tumors (n=100) were analyzed by ELISA. Subsequently, we validated our findings by analyzing an independent validation cohort comprising n=365 urothelial carcinoma urines. RESULTS: We found a statistically significant higher expression of Mxi-2 and Vim3 in patients afflicted with MIBC (p<0.001) compared to NMIBC. For Mxi-2 a sensitivity of 92% and specificity of 88% and for Vim3 a sensitivity of 82% and specificity of 84% could be determined in our cohort from the University Hospital Cologne. Moreover, in the independent validation cohort Mxi-2 determination showed a sensitivity of 80% and a specificity of 94%, while Vim3 showed a specificity of 80% and a sensitivity of 75%. The evaluation of Mxi-2 and Vim3 in patient urine achieves better sensitivity and specificity compared to existing urine test system for bladder cancer such as NMP22, BTA stat and Uromonitor. CONCLUSIONS: The determination of Mxi-2 and Vim3 in urine enables rapid and cost-effective differentiation between NMIBC and MIBC with a high diagnostic accuracy for tailored surgical and therapeutic evaluation. Prospective validation of urine-based prediction of muscle invasiveness is ongoing in the prospective, multicenter Bladder BRIDGister study. In addition, biomarker results are being compared with multiparametric MRI to provide an integrated, non-invasive platform for robust MIBC classification prior to surgery. Source of Funding: None © 2024 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 211Issue 5SMay 2024Page: e229 Advertisement Copyright & Permissions© 2024 by American Urological Association Education and Research, Inc.Metrics Author Information Barbara Koeditz More articles by this author Lucas Kastner More articles by this author Constantin Rieger More articles by this author Sofia Semko More articles by this author Maksyn Pikul More articles by this author Enno Storz More articles by this author Throsten Ecke More articles by this author Friederike Linden More articles by this author Ralph Wirtz More articles by this author Richard Weiten More articles by this author Melanie von Brandenstein More articles by this author Axel Heidenreich More articles by this author Expand All Advertisement PDF downloadLoading ...

545 Background: Patients with muscle invasive urothelial carcinoma achieving pathological complete response (pCR) upon neoadjuvant chemotherapy (NACT) have improved prognosis. Molecular subtypes of bladder cancer differ markedly with regard to sensitivity to cisplatinum based chemotherapy. Previously we did show that luminal tumors respond better to NACT, while FGFR1 expression is associated with chemo resistance (Ecke et al. 2022). The objective of this study was to determine which patients may benefit from Antibody Drug Conjugate (ADC) treatment in addition to NACT to justify subsequent prospective analysis within the "Bladder BRIDGister". Methods: Formalin fixed paraffin embedded (FFPE) tissues from transurethral resections (TUR) before chemotherapy and cystectomy samples after NACT of 36 patients were retrospectively collected. RNA from FFPE tissues were extracted by commercial kits, relative gene expression of subtyping markers (KRT5, KRT20, FGFR1) and radioligand target genes (NECTIN4, TROP2) were analyzed by standardized RT-qPCR systems (STRATIFYER Molecular Pathology GmbH, Cologne). Spearman correlation, hierarchical clustering, Kruskal-Wallis, chi square and contingency tests were done by JMP 9.0.0 (SAS software). Results: The neoadjuvant cohort consisted of 36 patients (median age: 69, male 83% vs. female 17%) with 92% of patients being pathohistologically node negative. When comparing pretreatment with post treatment samples the median expression of KRT20 dropped 128fold, while FGFR1 expression increased 6.8 fold. Interestingly, TROP2 and NECTIN4 mRNA expression also dropped significantly upon NACT by 5.7 fold and 7.1 fold, respectively. TROP2 and NECTIN4 were positively associated with the response marker KRT20 in therapy naïve TUR biopsies (r=0.5562 p=0.0004; r=0.5833 p=0.0002), but negatively associated with the resistance marker FGFR1 (r=-0.2903 p=0,0858; r=-0.3396 p=0,0427). However, TROP2 and NECTIN4 were not associated with pCR in spearman analysis with minor trend for TROP2 (r=0,2139 p=0,2103). Cluster analysis revealed a subgroup of KRT20 positive and FGFR1 negative tumors expression TROP2 and NECTIN4, which achieved 80% pCR. In addition elevated TROP2 and NECTIN4 expression was found in KRT20 positive tumors coexpressing FGFR1 and being resistant to NACT. Conclusions: Expression of the ADC targets TROP2 and NECTIN4 is associated with KRT20 positive, luminal tumors being highly sensitive to neoadjuvant chemotherapy alone. KRT5 positive, basal tumors do exhibit only very low expression of TROP2 and NECTIN4 mRNA. In view of toxicities the addition of TROP2 and NECTIN4 treatment to NACT might be considered only in luminal tumors exhibiting elevated FGFR1 expression as resistance mechanism and therefore do not respond to NACT.

Introduction and objective: BTA stat (R) , NMP22 (R) BladderChek (R) , UBC (R) Rapid Test, and CancerCheck (R) UBC (R) rapid VISUAL are uri-nary-based rapid tests. This multicenter study is the first study comparing all available rapid tests on a large cohort of bladder cancer patients and healthy controls in one setting. Methods: In total 732 urine samples (second morning urine) in a real-world assessment have been analyzed. We evaluated clinical sam-ples from 464 patients with histologically confirmed urothelial tumors of the urinary bladder (17 solitary CIS, 189 low-grade, 187 high-grade nonmuscle invasive, 71 high-grade muscle invasive), 77 patients with No Evidence of Disease (NED), and from 191 healthy controls. Urine samples were analyzed by the BTA stat (R), NMP22 (R) BladderChek (R), UBC (R) Rapid Test point-of-care (POC) system using the concile Omega 100 POC reader, and CancerCheck (R) UBC (R) rapid VISUAL. Sensitivities and specificities were calculated by contingency analyses. Results: All investigated urinary markers detected more pathological concentrations in urine of bladder cancer patients compared to tumor-free patients. The calculated diagnostic sensitivities for BTA stat (R), NMP22 (R) BladderChek (R), UBC (R) Rapid Test, CancerCheck (R) UBC (R) rapid VISUAL, and cytology were 62.4%, 13.4%, 58.2%, 28.6%, 36.2% for low-grade, 83.4%, 49.5%, 84.5%, 63.1%, 71.2% for high-grade nonmuscle invasive, and 95.8%, 35.2%, 76.1%, 50.7%, 67.7% for high-grade muscle-invasive bladder cancer. The specificity was 67.9%, 95.5%, 79.4%, 94.4%, and 83.7%, respectively. The area under the curve (AUC) after receiver operating characteristics (ROC) analysis for high-grade non-muscle-invasive tumors was 0.757, 0.725, 0.819, 0.787, and 0.774, respectively. Conclusions: The analysis of more than 700 urine samples offers an objective view on urine-based rapid diagnostics. Elevated patholog-ical concentrations of markers in urine of bladder cancer patients were detected in all investigated tests. The highest sensitivities for high-grade non-muscle-invasive tumors were calculated for BTA stat (R) and UBC (R) Rapid Test, whereas NMP22 (R) BladderChek (R), and cytology showed the highest specificities. BTA stat (R) and UBC (R) Rapid Test have the potential to be used as a clinical valuable urinary protein bio-marker for the detection of high-grade non-muscle-invasive bladder cancer patients and could be included in the management of these tumors. (c) 2023 Elsevier Inc. All rights reserved.

e16603 Background: Patients with muscle invasive urothelial carcinoma achieving pathological complete response upon neoadjuvant chemotherapy (NACT) have improved prognosis. Previously we did show that luminal tumors respond better to NACT, while FGFR1 expression is associated with NACT resistance (Ecke et al. 2022). Interestingly the expression of the radioligand targets CXCR4 and FAP is found in chemoresistant, stroma-associated tumors. The objective of this study was to prospectively validate the predictive value of molecular target typing from TUR biopsy samples and compare PET CT imaging by FDG and FAP radioligand imaging in selected patients after two cycles of NACT to justify subsequent adaptive trial concepts within the "Bladder BRIDGister." Methods: Formalin fixed paraffin embedded tissues (FFPE) from transurethral resections (TUR) before chemotherapy and cystectomy samples after NACT of 36 patients were retrospectively collected and 650 TURB samples were prospectively collected as part of the Bladder BRIDGister. RNA from FFPE tissues were extracted by commercial kits, relative gene expression of subtyping markers (KRT5, KRT20, FGFR1) and radioligand target genes (CXCR4, FAP) were analyzed by standardized RT-qPCR systems (STRATIFYER Molecular Pathology GmbH, Cologne). PET CT Imaging by FDG and FAP was performed after two cycles cisplatinum based NACT. Results: The neoadjuvant cohort consisted of 36 patients (median age 69, male 83%, female 17%) with 92% of patients being node negative. Hierarchical clustering revealed CXCR4 and FAP to be elevated in stromal rich, KRT5 & KRT20 negative tumors not responding to NACT. Elevated FAP mRNA expression was sig. associated with resistance to NACT (chi 2 4.314 p=0,0378). Combining elevated FAP and CXCR4 mRNA expression did identify 1/3 of the patients to be at high risk of NACT resistance (90%). Exemplarily one pT3 G3 patient was selected for PET CT imaging after two cycles that was predicted to be unresponsive to NACT by molecular subtyping. FDG PET CT revealed a hepatic metastatic lesion. In contrast FAP PET CT indicated multiple hepatic and a pancreatic metastatic lesion indicating tumor progression under NACT. Therapy was switched to MVAC with persistent non response. Then pembrolizumab monotherapy was administered due to PD-L1 positivity of the initial TURB (10% TPS /CPC 15). However there still was fulminant progression of the liver metastasis so that ICI therapy had to be stopped. Conclusions: Expression of the radioligand targets CXCR4 and FAP has been shown to be associated with aggressive stromal associated tumors and being resistant to NACT. This could be validated by selecting patients after two cycles of NACT for PET/CT imaging. Stratified FAP PET/CT turned out to be more sensitive than conventional FDG PET CT and may enable future theranostic treatment combinations in patients unresponsive to standard chemo- or immunotherapies.

Patients with muscle-invasive urothelial carcinoma achieving pathological complete response (pCR) upon neoadjuvant chemotherapy (NAC) have improved prognosis. Molecular subtypes of bladder cancer differ markedly regarding sensitivity to cisplatin-based chemotherapy and harbor FGFR treatment targets to various content. The objective of the present study was to evaluate whether preoperative assessment of molecular subtype as well as FGFR target gene expression is predictive for therapeutic outcome—rate of ypT0 status—to justify subsequent prospective validation within the “BladderBRIDGister”. Formalin-fixed paraffin-embedded (FFPE) tissue specimens from transurethral bladder tumor resections (TUR) prior to neoadjuvant chemotherapy and corresponding radical cystectomy samples after chemotherapy of 36 patients were retrospectively collected. RNA from FFPE tissues were extracted by commercial kits, Relative gene expression of subtyping markers (e.g., KRT5, KRT20) and target genes (FGFR1, FGFR3) was analyzed by standardized RT-qPCR systems (STRATIFYER Molecular Pathology GmbH, Cologne). Spearman correlation, Kruskal–Wallis, Mann–Whitney and sensitivity/specificity tests were performed by JMP 9.0.0 (SAS software). The neoadjuvant cohort consisted of 36 patients (median age: 69, male 83% vs. female 17%) with 92% of patients being node-negative during radical cystectomy after 1 to 4 cycles of NAC. When comparing pretreatment with post-treatment samples, the median expression of KRT20 dropped most significantly from DCT 37.38 to 30.65, which compares with a 128-fold decrease. The reduction in gene expression was modest for other luminal marker genes (GATA3 6.8-fold, ERBB2 6.3-fold). In contrast, FGFR1 mRNA expression increased from 33.28 to 35.88 (~6.8-fold increase). Spearman correlation revealed positive association of pretreatment KRT20 mRNA levels with achieving pCR (r = 0.3072: p = 0.0684), whereas pretreatment FGFR1 mRNA was associated with resistance to chemotherapy (r = −0.6418: p < 0.0001). Hierarchical clustering identified luminal tumors of high KRT20 mRNA expression being associated with high pCR rate (10/16; 63%), while the double-negative subgroup with high FGFR1 expression did not respond with pCR (0/9; 0%). Molecular subtyping distinguishes patients with high probability of response from tumors as resistant to neoadjuvant chemotherapy. Targeting FGFR1 in less-differentiated bladder cancer subgroups may sensitize tumors for adopted treatments or subsequent chemotherapy.