Primary Urothelial Carcinoma Research Articles

An Unordinary Recurrence of Urothelial Bladder Carcinoma in the Colon Presenting as Hematochezia

The conventional route of recurrence and metastasis in primary urothelial cell carcinoma of the bladder is through lymphatics and hematogenous system with regional lymph nodes, lungs, liver, brain and bone being the most commonly involved sites. Invasive urothelial cancers could also spread locoregionally to surrounding organs including female genital tract, prostate and gastrointestinal tract especially colo-rectum. However, recurrence of this cancer inside the colon in the absence of locoregional invasion, and after achieving complete long-term remission is extremely unusual. A review of the literature revealed a single case reported by Yang et al. of an urothelial cancer recurrence in the colon with mixed urothelial/colonic malignant pathology [1]. Herein we report the second case, to our knowledge, of an urothelial cell bladder carcinoma that recurred inside the colon in the form of a mass causing hematochezia, which on pathology and immunohistostaining proved to be primarily of urothelial rather than colonic origin. 67 year old male heavy smoker with stage III (T3a, N0, M0) transitional cell carcinoma of the bladder had undergone radical cystectomy, neo-bladder reconstruction and adjuvant chemotherapy following initial diagnosis. He poved repetitively to have no evidence of disease until he presented 4 years later with intermittent hematochezia and anemia. Colonoscopy revealed a fungating, ulcerated mass in the sigmoid colon suggestive of a primary colon cancer (fig 1). Pathology disclosed high grade/poorly differentiated carcinoma invading the colonic mucosa that surprisingly favored an urothelial origin. Upon further immunostaining, final diagnosis of a metastatic high-grade urothelial carcinoma was confirmed by positive cytokeratin (CK) 7 stain (specific for urothelial cells) and negative villin and CDX2 stains that are both specific for gastrointestinal differentiation (fig 2). CT scan of abdomen and pelvis, and PET scan did not reveal any other sites of cancer recurrence. Patient received a tailored chemotherapy and responded appropriately.Figure 1Figure 2This case illustrates a rare kind of recurrence of urothelial cancer in the form of a colonic mass resembling primary colon cancer. This unordinary way of recurrence could pose a clinico-pathological diagnostic challenge in distinguishing this disease treated basically with chemotherapy, from a primary colon cancer managed primarily by surgical resection. Immunohistochemical stains are essential for an accurate diagnosis. Heightened awareness is of utmost importance in such patients with history of urothelial cancer in order to appropriately tailor further treatment strategies that differ between primary and secondary colonic tumors.

A cultured approach to canine urothelial carcinoma: molecular characterization of five cell lines.

BackgroundUrothelial carcinoma (UC), also known as transitional cell carcinoma (TCC), of the bladder is the most common neoplasm affecting the canine urogenital system. To facilitate study of the disease in vitro, cell line models have been established from primary tumor biopsies. Their resemblance to the primary disease, however, has not been well defined. In the present study, we evaluated five canine UC cell lines via oligonucleotide array comparative genomic hybridization (oaCGH), fluorescence in situ hybridization (FISH), and gene expression analysis.ResultsComparison of genome wide DNA copy number profiles of the cell lines with primary biopsy specimens revealed redundancies in genomic aberrations, indicating that the cell lines retain the gross genomic architecture of primary tumors. As in the primary tumors, gain of canine chromosomes 13 and 36 and loss of chromosome 19 were among the most frequent aberrations evident in the cell lines. FISH analysis revealed chromosome structural aberrations, including tandem duplications, bi-armed chromosomes, and chromosome fusions, suggesting genome instability during neoplastic transformation. Gene expression profiling highlighted numerous differentially expressed genes, including many previously shown as dysregulated in primary canine UC and human bladder cancer. Pathway enrichment analysis emphasized pathways suspected to be at the crux of UC pathogenesis, including xenobiotic and lipid compound metabolism.ConclusionsThese data support valid use of the canine UC cell lines evaluated by confirming they provide an accurate and practical means to interrogate the UC at a molecular level. Moreover, the cell lines may provide a valuable model for furthering our understanding of aberrant metabolic pathways in UC development.Electronic supplementary materialThe online version of this article (doi:10.1186/s40575-015-0028-3) contains supplementary material, which is available to authorized users.

Managing noninvasive recurrences after definitive treatment for muscle-invasive bladder cancer or high-grade upper tract urothelial carcinoma.

Approximately 50% of patients with muscle invasive urothelial carcinoma will relapse with distant recurrence. Though rates of local recurrence after definitive therapy have improved, management remains a challenge. In this review, treatment strategies for this cohort are re-examined in an effort to enhance patient outcomes. Urothelial carcinoma continues to demonstrate high rates of recurrence and low rates of survival. Similarly to the treatment of primary urothelial cancer, treatment of recurrence focuses on cytology, stage, and clinical characteristics. Current areas of interest have focused on identification and causes/predictors of recurrence. Limited progress has been achieved in differentiating management of recurrent urothelial carcinoma from the treatment of primary urothelial carcinoma. However, there may be an increasing role for endoscopic and organ conserving therapies for carefully selected patients with recurrent noninvasive urothelial carcinoma. Identifying those at risk for early recurrence and early diagnosis of recurrence may be the most beneficial future strategies. The treatment regimen for noninvasive bladder recurrence after radical nephroureterectomy for upper tract urothelial carcinoma should include intravesical chemotherapy or Bacillus Calmette-Guerin to prevent further bladder recurrence or tumor progression. We do not advocate diversion sparing techniques for local recurrence after radical cystectomy. Metastasectomy for distant/metastatic urothelial carcinoma recurrence represents a promising area of future study.

Connexin 43 expression predicts poor progression-free survival in patients with non-muscle invasive urothelial bladder cancer

ObjectivesTo evaluate the protein expression of connexin 43 (Cx43) in primary urothelial bladder cancer and test its association with the histopathological characteristics and clinical outcome.MethodsA tissue microarray containing 348 tissue...

Abstract 4943: Engulfment gene GULP1 is a functional tumor suppressor through influencing TGF-β pathway and is silenced by promoter methylation in urothelial carcinoma

Abstract GULP1 functions as a cytoplasmic adaptor protein in the engulfment of apoptotic corpses and cells. It was also reported to be an adapter protein of LRP1 which is known to be a key regulator of TGF-β signaling. TGF- β signaling pathway is known to contribute to urothelial carcinoma (UC) development by increasing invasiveness and metastasis, and inducing epithelial to mescenchymal (EMT) transition. By an integrated genetic approach, we recently identified GULP1 as a potential tumor suppressor gene and epigenetically silenced in ovarian cancer. In the present study we evaluated the promoter methylation of GULP1 in urothelial carcinoma (UC) and functionally characterize GULP1 in urothelial carcinogenesis. We first determined GULP1 protein expression by immunohistochemistry on a customized muscle-invasive UC (MIUC) tissue-microarray. The mechanism of gene silencing was probed using quantitative methylation-specific PCR (QMSP). The association between GULP1 expression and promoter methylation was analyzed by using a subset of primary UC and UC cell lines. Effects of forced overexpression and siRNA-mediated GULP1-knockdown were measured by cell proliferation and colony forming assays. Western blot analysis was performed for several key molecules to determine the effect of GULP1 modulation in the respective pathways. Complete loss of GULP1 expression was observed in 89 out of 104 MIUC (85.6%). In primary tumor tissues and cell lines, promoter methylation of GULP1 were inversely correlated with GULP1 expression and reactivation of GULP1 was observed by 5-aza-dC treatment of UC cell lines. Additionally QMSP assay of GULP1 was performed for 98 primary UC tissues of different cell types and 21 normal urothelial samples. Our initial analysis by using an empiric cut off value revealed GULP1 promoter methylation was significantly associated with UC (P = 0.007, Fisher's exact test). To determine the feasibility of GULP1 QMSP assay for non-invasive detection of UC using urine DNA, we tested urine from 58 UC patients and 46 healthy people. As expected, 33 out of 58(56.9%) UC cases were methylation positive while very low level of methylation were detected in 4 out of 46 urine control samples (8.7%) (P<0.001, Fisher's exact test). Effect of in vitro modulation of GULP1 was determined by MTT assay, BrdU assay and colony formation assay. The results consistently revealed that GULP1 had anti-proliferative activities. In order to detect the potential pathway related to GULP1, RNA from T24 and shGULP1 transfected T24 cells were applied to Cancer pathway finder array (Qiagen). Heme oxygenase 1 (HMOX1) solely showed over 3 fold increase in T24shGULP1 cells. Other associated molecules altered due to GULP1 knock-down include TGF-β RI, p-Smad3C, p15 and LRP1. In conclusion, GULP1 is a candidate tumor suppressor in UC carcinogenesis and it may have a potential for non-invasive UC detection and targeting therapy. Citation Format: Masamichi Hayashi, Leonardo O. Reis, Alexander Baras, Leonel Maldonado, Eliza Guida, Evgeny Izumchenko, Mariana Brait, Trinity Bivalacqua, George J. Netto, Wayne Koch, David Sidransky, Mohammad O. Hoque. Engulfment gene GULP1 is a functional tumor suppressor through influencing TGF-β pathway and is silenced by promoter methylation in urothelial carcinoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4943. doi:10.1158/1538-7445.AM2015-4943

Impact of reduced levels of APE1 transcripts on the survival of patients with urothelial carcinoma of the bladder.

Molecular evidence indicates that alterations in genes involved in the maintenance of genome stability may be related to susceptibility to bladder carcinoma. Our goal was to evaluate the prognostic role of base excision repair (BER) genes in a cohort of patients diagnosed with primary urothelial carcinoma of the bladder (UCB). The levels of all APE1, XRCC1 and POLB transcripts were detected by quantitative real-time PCR (qPCR) technique in tumor samples from 52 patients undergoing transurethral resection (TUR) for primary UCB at the Department of Urology, Brazilian National Cancer Institute, Rio de Janeiro. Increased levels of APE1, XRCC1 and POLB transcripts were significantly associated with high-grade tumors when compared to these levels in low-grade tumors (p<0.01) and could be attributed to different mechanisms of transcriptional regulation as a response to tumorigenesis and oxidative stress. By analyzing the collected data in the present study, regardless of pathological grade or stage, univariate analysis revealed that the reduced levels of APE1 transcripts were significantly associated with cancer-specific mortality (p=0.032). Furthermore, the variant genotype (TG/GG) of the APE1 T1349G polymorphism was observed in 75% of a subset of patients who concomitantly experienced reduced levels of the APE1 transcript and death and/or recurrence events. Taken together, our data reinforce the idea that human DNA repair mechanisms must be finely regulated in order to avoid instability leading to tumorigenesis and poor clinical outcomes in UCB patients.

Risk factors analysis and prognosis of bladder carcinoma occurrence in patients with transitional-cell carcinoma of primary upper urinary tract after radical operation

Objective To analysis the risk factors and prognosis of bladder carcinoma currence in patients with transitional-cell carcinoma of primary upper urinary tract after radical operation. Methods The clinical data were retrospectively collected in 115 patients with carcinoma of primary upper tract. The protein expressions of tumor markers Ki-67 and P53 were detected by immunohistochemistry. Combined with traditional cliniopathological factors and molecular markers, the risk factors of bladder carcinoma occurrence were analyzed. Results The incidence of bladder cancer was 11.3%(13/115) and the mean follow-up time was 48.5 months. Age more than 65 years (P=0.040), ureteral tumors (P=0.008) and the lower expression of Ki-67 (P=0.041) were independent predictors for bladder cancer. The survival time of non-progression were (92.1±13.3) and (112.8±4.3) months in bladder tumor group and non-bladder tumor group, but there was no significant difference between the two groups (P>0.05). The survival time of no disease were (113.5±4.3) and (116.0±5.0) months, respectively (P>0.05). Conclusions Advance age, lower ureteral tumors and down-expression of Ki-67 were independent predictors for bladder tumor recurrence in primary upper tract urothelial carcinomas after nephroureterectomy. There was no relationship between bladder tumor recurrence and oncological outcomes. However, close follow-up and rigorous treatment are essential for patients with high risk factors. Key words: Carcinoma, transitional cell; Urinary Bladder neoplasms; Prognosis

Immunohistochemical assessment of lymphatic and blood vessel invasion in T1 urothelial carcinoma of the bladder

Objective. The aim of this study was evaluate the incidence and significance of immunohistochemically assessed lymphatic (LVI) and blood vessel invasion (BVI) in primary T1 urothelial carcinoma of the bladder (UCB) treated with radical cystectomy (RC). Materials and methods. Thirty-two patients with T1 UCB at primary diagnosis were identified who underwent radical cystectomy (RC) subsequently. Of these, 16 (50%) had pT1N0M0 (group I) and 16 (50%) ≥ pT2aN0-3M0 UCB (group II) at RC. The presence of LVI and BVI in transurethral resection of bladder tumor (TURBT) and corresponding RC specimens was assessed using hematoxylin & eosin (H&E) and immunohistochemical (IHC) staining against the lymphatic (D2-40) and vascular endothelium (CD31). Results. At TURBT and RC, none of the patients in group I showed LVI or BVI on H&E and IHC sections. In group II, at TURBT, LVI and BVI were negative on H&E staining in all patients, but detectable by IHC in two patients (13%) and one patient (6%), respectively (p = 0.48 and p = 0.99 compared to group I). At RC, LVI and BVI were detected by IHC in eight (50%) and five (31%) of the 16 patients, respectively (p = 0.002 and p = 0.021 compared to group I). Of these eight and five patients, detection of LVI and BVI was only possible with IHC in six (75%) and three (60%), respectively. Conclusions. Although this hypothesis-generating study did not show a high degree of concordance between TURBT and RC specimens, IHC assessment on a regular basis may increase the detection rates of LVI and BVI at initial diagnosis and improve the selection of those T1 patients who should be offered early radical treatment.

Penile-preserving surgery for primary urothelial carcinoma of male urethra

Primary urethral carcinoma is a rare cancer, comprising <1% of all malignancies. The location of this lesion presents a certain dilemma of treatment between efficacy and quality of life. We report an 84-year-old male patient, with a history of chronic hepatitis C, hypertension, and transient ischemic accident, who presented with dysuria and acute urinary retention. The intravenous urography showed mild prostatic enlargement, but no stone or filling defect was noted in the upper urinary tract. On urethrocystoscopy, multiple papillary tumors were found at the pendulous urethra, and the pathology of biopsy confirmed urothelial carcinoma. The patient was admitted, and electroresection with fulguration of urethral tumors was performed owing to the patient's old age and poor performance status. Intraurethral and intravesical chemotherapy with mitomycin C was regularly given at the outpatient clinic. Recurrent urothelial carcinomas were noted twice in the first 2 years of follow up, and repeated transurethral resections were done. Unfortunately, liver cirrhosis with hepatocellular carcinoma was diagnosed last June, for which he received transcatheter arterial chemoembolization. No recurrence of urethral cancer has been found on semiannual cystoscopy in the past 3 years. Penile-preserving surgery is a reasonable surgical option for elderly primary urethral carcinoma patients with acceptable oncological outcome and good quality of life.

DNA copy number analysis of metastatic urothelial carcinoma with comparison to primary tumors.

BackgroundTo date, there have been no reports characterizing the genome-wide somatic DNA chromosomal copy-number alteration landscape in metastatic urothelial carcinoma. We sought to characterize the DNA copy-number profile in a cohort of metastatic samples and compare them to a cohort of primary urothelial carcinoma samples in order to identify changes that are associated with progression from primary to metastatic disease.MethodsUsing molecular inversion probe array analysis we compared genome-wide chromosomal copy-number alterations between 30 metastatic and 29 primary UC samples. Whole transcriptome RNA-Seq analysis was also performed in primary and matched metastatic samples which was available for 9 patients.ResultsBased on a focused analysis of 32 genes in which alterations may be clinically actionable, there were significantly more amplifications/deletions in metastases (8.6% vs 4.5%, p < 0.001). In particular, there was a higher frequency of E2F3 amplification in metastases (30% vs 7%, p = 0.046). Paired primary and metastatic tissue was available for 11 patients and 3 of these had amplifications of potential clinical relevance in metastases that were not in the primary tumor including ERBB2, CDK4, CCND1, E2F3, and AKT1. The transcriptional activity of these amplifications was supported by RNA expression data.ConclusionsThe discordance in alterations between primary and metastatic tissue may be of clinical relevance in the era of genomically directed precision cancer medicine.Electronic supplementary materialThe online version of this article (doi:10.1186/s12885-015-1192-2) contains supplementary material, which is available to authorized users.

MP68-08 CONNEXIN 43 EXPRESSION PREDICTS POOR PROGRESSION-FREE SURVIVAL IN PATIENTS WITH NON-MUSCLE INVASIVE UROTHELIAL BLADDER CANCER

You have accessJournal of UrologyBladder Cancer: Basic Research IV1 Apr 2015MP68-08 CONNEXIN 43 EXPRESSION PREDICTS POOR PROGRESSION-FREE SURVIVAL IN PATIENTS WITH NON-MUSCLE INVASIVE UROTHELIAL BLADDER CANCER Cédric Poyet, Lorenz Buser, Filip Roudnicky, Michael Detmar, Thomas Hermanns, Doris Mannhard, Tullio Sulser, Holger Moch, and Peter Wild Cédric PoyetCédric Poyet More articles by this author , Lorenz BuserLorenz Buser More articles by this author , Filip RoudnickyFilip Roudnicky More articles by this author , Michael DetmarMichael Detmar More articles by this author , Thomas HermannsThomas Hermanns More articles by this author , Doris MannhardDoris Mannhard More articles by this author , Tullio SulserTullio Sulser More articles by this author , Holger MochHolger Moch More articles by this author , and Peter WildPeter Wild More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2015.02.2470AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Connexins (Cxs) which are considered to compose gap junctions to form intercellular channels, play an important role in the regulation of proliferation, cell growth and apoptosis. One of the most studied Cx proteins is connexin 43 (Cx43). There is evidence that Cx43 is involved in cancer development and metastatic process and overexpressed in invasive lesions of some solid tumours like breast or colon cancer. The objective of the study was to evaluate the protein expression of Cx43 in primary urothelial bladder cancer and to test its association with histopathological characteristics and clinical outcome. METHODS A tissue microarray containing 348 tissue samples from 174 patients with primary urothelial carcinomas of the bladder was immunohistochemically stained for Cx43. Intensity of staining was evaluated semiquantitatively (score 0, 1+, 2+), and the association with clinico-pathological features was assessed. Univariate and multivariate analyses were performed to identify predictors for progression-free survival (PFS). RESULTS Membranous Cx43 immunoreactivity was detected in 118 (67.8 %) of 174 analyzable urothelial carcinomas, of which 31 (17.8%) showed even a strong (score 2+) and mainly homogeneous staining. Strong expression levels of Cx43 (score 2+) were associated with higher tumor grade, multiplicity and increased Ki-67 proliferation (all p < 0.05). In the subgroup of stage pTa and pT1 bladder tumours (n = 158), strong Cx43 expression (p < 0.001), solid growth pattern (p < 0.001) and increased proliferation fraction (p < 0.05) were significantly associated with shorter PFS in a univariate Cox regression analysis. In multivariate Cox regression models, Cx43 immunoreactivity and histologic growth pattern remained highly significant, independent, and adverse risk factors for PFS. CONCLUSIONS Expression levels of Cx43 are frequent in non-muscle invasive bladder cancer (NMIBC), with high expression levels beeing associated with poor prognosis. Routine assessment of Cx43 expression may improve the identification of high-risk NMIBC. © 2015 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 193Issue 4SApril 2015Page: e860-e861 Advertisement Copyright & Permissions© 2015 by American Urological Association Education and Research, Inc.MetricsAuthor Information Cédric Poyet More articles by this author Lorenz Buser More articles by this author Filip Roudnicky More articles by this author Michael Detmar More articles by this author Thomas Hermanns More articles by this author Doris Mannhard More articles by this author Tullio Sulser More articles by this author Holger Moch More articles by this author Peter Wild More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Canine urothelial carcinoma: genomically aberrant and comparatively relevant.

Urothelial carcinoma (UC), also referred to as transitional cell carcinoma (TCC), is the most common bladder malignancy in both human and canine populations. In human UC, numerous studies have demonstrated the prevalence of chromosomal imbalances. Although the histopathology of the disease is similar in both species, studies evaluating the genomic profile of canine UC are lacking, limiting the discovery of key comparative molecular markers associated with driving UC pathogenesis. In the present study, we evaluated 31 primary canine UC biopsies by oligonucleotide array comparative genomic hybridization (oaCGH). Results highlighted the presence of three highly recurrent numerical aberrations: gain of dog chromosome (CFA) 13 and 36 and loss of CFA 19. Regional gains of CFA 13 and 36 were present in 97 %and 84% of cases, respectively, and losses on CFA 19 were present in 77% of cases. Fluorescence in situ hybridization (FISH), using targeted bacterial artificial chromosome (BAC) clones and customAgilent SureFISH probes, was performedto detect and quantify these regions in paraffin-embedded biopsy sections and urine-derived urothelial cells. The data indicate that these three aberrations are potentially diagnostic of UC. Comparison of our canine oaCGH data with that of 285 human cases identified a series of shared copy number aberrations. Using an informatics approach to interrogate the frequency of copy number aberrations across both species, we identified those that had the highest joint probability of association with UC. The most significant joint region contained the gene PABPC1, which should be considered further for its role in UC progression. In addition, cross-species filtering of genome-wide copy number data highlighted several genes as high-profile candidates for further analysis, including CDKN2A, S100A8/9, and LRP1B. We propose that these common aberrations are indicative of an evolutionarily conserved mechanism of pathogenesis and harbor genes key to urothelial neoplasia, warranting investigation for diagnostic, prognostic, and therapeutic applications.

A Molecular Pathologic Framework for Risk Stratification of Stage T1 Urothelial Carcinoma

BackgroundOne third of patients with stage T1 urothelial carcinoma (UC) progress to muscle-invasive disease requiring radical surgery. Thus, reliable tools are needed for risk stratification of stage T1 UC. ObjectiveTo investigate the extent to which stratification of stage T1 tumours into previously described molecular pathologic UC subtypes can provide improved information on tumour progression. Design, setting, and participantsA population-based cohort of 167 primary stage T1 UCs was characterised by immunohistochemistry and classified into the molecular subtypes urobasal (Uro, 32%), genomically unstable (GU, 58%), and squamous-cell-carcinoma–like (SCCL, 10%). Outcome measurements and statistical analysisProgression-free survival using univariate and multivariate models. Results and limitationsSubtype classification was validated using nine additional markers with known subtype-specific expression. Analysis of mRNA expression of progression biomarkers revealed a strong association with molecular subtype. Kaplan-Meier analyses showed that the risk of progression was low for Uro tumours and high for GU/SCCL tumours. High progression risk scores were found only for GU/SCCL tumours. Clinical risk factors such as multifocality, concomitant carcinoma in situ, invasion depth, lymphovascular invasion, and high CD3+ lymphocyte infiltration were observed almost exclusively in GU/SCCL cases. ConclusionsMolecular subtypes Uro, GU, and SCCL were identified in an independent population-based cohort of stage T1 UCs. Biomarkers and clinical risk factors for progression were associated with molecular subtype. Rapidly progressing T1 tumours were of subtype GU or SCCL and had either a high progression risk score or an elevated CD3+ cell count. Patient summaryWe show that classification of stage T1 urothelial carcinoma into molecular subtypes can improve the identification of patients with progressing tumours.

Clonality of bladder tumors following radical nephroureterectomy: Against the field defect hypothesis.

301 Background: Urothelial carcinoma is characterized by multifocal and metachronous tumors. To explain this phenomenon, two hypotheses have been proposed: the ‘field defect’ hypothesis - urothelial cells are primed to undergo transformation by exposure to carcinogens, and the clonal, or ‘single progenitor cell,’ hypothesis - tumors arise from intraluminal seeding of transformed cells. Methods: To examine their clonal relationships, we compared the genomic profiles of primary upper tract urothelial carcinoma (UTUC) tumors and metachronous bladder tumors (intravesical recurrences) in patients treated with radical nephroureterectomy (RNU) and subsequent transurethral resection. Specimens were analyzed using a next-generation, targeted sequencing assay designed to identify point mutations, indels, and copy number alterations in 341 cancer-associated genes. Results: We analyzed 16 primary UTUC tumors and 41 intravesical recurrences in patients treated with RNU. The median number of intravesical recurrences per patient was 2 (range 1-7) and the interval from RNU to intravesical recurrence ranged from 3.5 months to 129 months. With an average sequencing coverage of 516x, we found strong evidence delineating the clonal relationship between primary UTUC tumors and subsequent bladder tumors. The majority of somatic mutations present in the primary UTUC tumors (median=7, range 4-39) were detected in all subsequent bladder tumors (128/146, 88%). In an illustrative case, one patient followed with periodic cystoscopy/cytology who had been NED for 5.5 years then developed 7 bladder tumors over the next 44 months, each with the identical mutation profile (8 mutations) as the primary tumor. Conclusions: We demonstrate that bladder tumors following RNU represent true intravesical recurrences, with almost all tumors sharing the same somatic mutation profile as the primary UTUC tumor. This has important implications for surgical techniques to minimize the risk of intraluminal seeding, the delivery of intravesical therapy following RNU, and the development of strategies employing systemic chemotherapy or targeted agents.

Postchemotherapy consolidative surgery for metastatic urothelial carcinoma.

362 Background: The role of post-chemotherapy consolidative surgery for metastatic urothelial carcinoma (UC) is controversial and supported by small, single-center series in select patients. We sought to assess our experience with post-chemo retroperitoneal lymph node (RPLN) dissection for primary metastatic or relapsed UC to the retroperitoneum (with or without concomitant cystectomy and/or nephroureterectomy). Methods: Since 2008, 15 patients with metastatic or relapsed UC of the bladder or upper tracts (UT) and associated RPLN metastases were considered for post-chemo consolidative surgery. Records were reviewed for response to therapy, final pathology, and impact of surgery on overall survival (OS). Chemo agents used were provider dependent with 9 patients treated with gemcitabine/cisplatin, 3 with gem/carboplatin, and 2 with methotrexate, vinblastine, doxorubicin, cisplatin (1 received a combination due to intolerance). A median of 3.5 cycles were given in the adjuvant (n=5) and induction (n=10) setting. Seven patients presented with UT disease, of those 3 with relapse; median follow-up was 31.6 months (mos, IQR 14, 72), with 1 patient deceased. Nine patients had a bladder primary, 5 had induction chemo (median follow-up: 22 mos, IQR 16, 40), with 1 alive, 4 dead of disease, and 2 of other causes. To date, 13 patients have undergone surgery. Overall median follow-up is 23 mos. Results: Five patients (3 bladder, 2 UT) were noted to have radiographic complete response (CR) with induction and a median survival of 21.5 mos, with one patient alive at 7 years. All but one patient in the UT group are alive. Two patients had pathological CR, both were in the bladder cohort and died at 8 and 23 mos post-op. For all other patients, final pathology noted a median of 2 LN positive for malignancy (average 3.6). Neither chemo nor timing of administration showed a difference in OS. Conclusions: There is a role for post-chemo consolidative surgery in selected patients with metastatic UC. Compared to patients with the primary tumor in the UT, those with a primary bladder tumor and RPLN fared worse with a shorter overall survival time. More studies are necessary to better define the role of surgery in this setting.

Genomic landscape of metastatic platin-resistant urothelial cancer patients.

316 Background: Although several studies have reported genomic characterisation of primary urothelial carcinoma (UC), little is known about the genomic alterations of the metastatic cisplatin-resistant UC. We sought to define the prevalence and co-occurrence of actionable genomic alterations in patients with metastatic cisplatin-resistant lethal UC. Methods: Patients with metastatic UC, who progressed after at least one line of standard therapy, were enrolled in a prospective molecular screening trial (MOSCATO 01) at Gustave Roussy. CT-Scan or ultrasound-guided biopsies were performed in metastases to carry out a comprehensive molecular characterization. DNA was extracted from fresh tumor samples and analyzed by comparative genomic hybridization (CGH) (≥ 30% tumor cells) and by Next Generation Sequencing (NGS) for up to 74 target genes (≥ 10% tumor cells). Whole-exome (WES) and RNA seq sequencing were performed retrospectively in selected cases. Results: From 12/2011 to 12/2013, 30 heavily pretreated patients with metastatic UC were included. Median age was 61 (37-73); all patients had been treated with platin-based chemotherapy with a median number of lines of 2.5 (1-5). A tumor biopsy could be performed in 26 patients (87%). CGH and targeted exome sequencing profiles were assessed in 19 (73%) and 23 (88%) of them, respectively. A total of 19 patients (73%) were profiled for both sequencing and CGH. WES and RNA seq was performed in 16 (62%) pairs of metastatic tissue and normal DNA (Integragen Inc, Hiseq platform). Our analyses identified potential therapeutic targets in 61 % of the tumours, including 31% with targets in the PI3K/AKT/mTOR pathway, 35% with targets in the FGF/FGFR pathway and 15% with targets in the RTK/MAPK pathway (including ERBB2). Others frequent aberrations were found in the chromatin regulatory genes (MLL gene family) and cell-cycle regulatory genes (E2F3, CDKN2A/B genes). FGFR gene fusions were found in 1 out of 16 screened patients (6%) Conclusions: Lethal platin-resistant UC exhibit high-level of genomic heterogeneity. The majority of these tumors harbour actionable genomic alterations that can be targeted with selective agents currently in clinical development (phase I trials) or registered.

CD73 Predicts Favorable Prognosis in Patients with Nonmuscle-Invasive Urothelial Bladder Cancer.

Aims. CD73 is a membrane associated 5′-ectonucleotidase that has been proposed as prognostic biomarker in various solid tumors. The aim of this study is to evaluate CD73 expression in a cohort of patients with primary bladder cancer in regard to its association with clinicopathological features and disease course. Methods. Tissue samples from 174 patients with a primary urothelial carcinoma were immunohistochemically assessed on a tissue microarray. Associations between CD73 expression and retrospectively obtained clinicopathological data were evaluated by contingency analysis. Survival analysis was performed to investigate the predictive value of CD73 within the subgroup of pTa and pT1 tumors in regard to progression-free survival (PFS). Results. High CD73 expression was found in 46 (26.4%) patients and was significantly associated with lower stage, lower grade, less adjacent carcinoma in situ and with lower Ki-67 proliferation index. High CD73 immunoreactivity in the subgroup of pTa and pT1 tumors (n = 158) was significantly associated with longer PFS (HR: 0.228; p = 0.047) in univariable Cox regression analysis. Conclusion. High CD73 immunoreactivity was associated with favorable clinicopathological features. Furthermore, it predicts better outcome in the subgroup of pTa and pT1 tumors and may thus serve as additional tool for the selection of patients with favorable prognosis.

Conservative management and female gender are associated with increased cancer-specific death in patients with isolated primary urothelial carcinoma in situ.

Our goal was to investigate the effect of patient and disease characteristics on the probability of cancer-specific death (CSD) in cases of isolated urothelial carcinoma in situ (CIS). We performed a retrospective analysis of patients diagnosed with isolated CIS between 1990 and 2010 identified from the Surveillance, Epidemiology, and End Results (SEER) database. Competing risk analysis using Cox proportional hazard model was used to examine the probability of CSD controlling for possible covariates. Overall (n = 1432), patients were mainly male (75%), mean age at diagnosis was 71 years, median survival 47 months, and 65% of the patients had CIS in their upper urinary tract. Caucasians were the predominant race (90%). CIS was the cause of death in 87/1432(6%) of the total cohort; 69/1239 (6%) of patients who underwent surgery, and 18/193 (9%) of the patients who were managed conservatively (CM). On multivariate analysis, CM [hazard ration (HR) = 2.019, CI: 1.189-3.429, P = 0.009] and female gender (HR = 1.690, CI: 1.041-2.741, P = 0.033) were associated with CSD, while age, site, race and year of diagnosis were non-significant predictors. Female gender and conservative management were positively associated with CSD. Multi-institutional collaboration is needed to validate markers for poor prognosis in cases of isolated CIS.

Expression of ProEx C in primary and metastatic urothelial carcinoma.

ProEx C is an antibody cocktail targeting the expression of topoisomerase IIα and minichromosome maintenance protein-2. Both these proteins are over-expressed in the cell nucleus during aberrant S-phase induction of neoplastic cells, which leads to cell proliferation. The aim of this study was to determine whether ProEx C expression can detect primary and metastatic urothelial carcinoma (UC). Thirty one fine needle aspiration cell blocks (CB) with metastatic UC were identified. Immunohistochemical staining for ProEx C and thrombomodulin was performed. Additionally, staining for Pro Ex C was also performed in tissue microarrays (TMA) of 46 cases of primary UC and carcinomas from colon (80), stomach (31), pancreas (33), liver (92), ovary (24), endometrium (25), breast (60), lung (27), kidney (32), and prostate (44), as well as melanoma (22). Nuclear staining of ProEx C and membrane staining of thrombomodulin in at least 10% tumor cells was considered a positive result. Both ProEx C and thrombomodulin have similar sensitivity for metastatic UC (84% vs. 77%, p=0.75; whereas ProEx C yielded a higher sensitivity of 93% for primary UC than thrombomodulin (72%, p=0.01). In addition to UC, ProEx C is also expressed in most of the malignant neoplasms tested in our TMA study, and has the highest sensitivity in colon and stomach carcinomas (94%). ProEx C has high sensitivity for UC. However, it is also expressed in carcinomas of colon, stomach, breast, and lung carcinomas and may not be a useful marker for workup of metastatic UC.

FGFR3 expression in primary and metastatic urothelial carcinoma of the bladder.

While fibroblast growth factor receptor 3 (FGFR3) is frequently mutated or overexpressed in nonmuscle-invasive urothelial carcinoma (UC), the prevalence of FGFR3 protein expression and mutation remains unknown in muscle-invasive disease. FGFR3 protein and mRNA expression, mutational status, and copy number variation were retrospectively analyzed in 231 patients with formalin-fixed paraffin-embedded primary UCs, 33 metastases, and 14 paired primary and metastatic tumors using the following methods: immunohistochemistry, NanoString nCounterTM, OncoMap or Affymetrix OncoScanTM array, and Gain and Loss of Analysis of DNA and Genomic Identification of Significant Targets in Cancer software. FGFR3 immunohistochemistry staining was present in 29% of primary UCs and 49% of metastases and did not impact overall survival (P = 0.89, primary tumors; P = 0.78, metastases). FGFR3 mutations were observed in 2% of primary tumors and 9% of metastases. Mutant tumors expressed higher levels of FGFR3 mRNA than wild-type tumors (P < 0.001). FGFR3 copy number gain and loss were rare events in primary and metastatic tumors (0.8% each; 3.0% and 12.3%, respectively). FGFR3 immunohistochemistry staining is present in one third of primary muscle-invasive UCs and half of metastases, while FGFR3 mutations and copy number changes are relatively uncommon.