Biology Of Bladder Cancer Research Articles

PCN273 EPIDEMIOLOGY AND UNMET NEEDS OF BLADDER CANCER IN ITALY

Bladder cancer is one of the most frequent cancers in high-income countries. Information on bladder cancer in Italy is scattered across scientific literature and institutional and educational resources, and no attempt has been made yet to organize and summarize this information across various sources of available data. The aims of this review are to summarize recent epidemiological data of bladder cancer (including incidence, prevalence, survival and mortality) and to evaluate patients’ unmet needs. We reviewed the scientific literature of the last 10 years through PubMed/Medline and Google Scholar and we deeply explored the grey literature. Available official statistics indicate a high burden of bladder cancer in Italy, where this neoplasm has one of the highest incidences worldwide and, in consideration of its relatively high survival, it ranks 4th in cancer prevalence, after neoplasms of the breast, prostate and colorectal. The limited therapeutic options for muscle-invasive and advanced/metastatic urothelial cancer are one of the major unmet needs for patients with this neoplasm, in Italy and worldwide. Advances in cancer immunotherapy and in understanding molecular biology of bladder cancer are, however, rapidly altering the therapeutic landscape for targeted subgroups of patients with advanced/metastatic disease. Other unmet needs include low quality of life after radical cystectomy, the lack of widespread clinical pathway schemes to improve and standardize quality of care and a low Italian patients’ empowerment. This review provides a summary of recent epidemiological and health care data of urothelial cancer, including patients’ unmet needs, with a focus on Italy, one of the countries with the highest incidence rate worldwide.

Connecting the dots between different networks: miRNAs associated with bladder cancer risk and progression

BackgroundBladder cancer (BC) is a common urothelial malignancy, characterized by a high recurrence rate. The biology of bladder cancer is complex and needs to be deciphered. The latest evidence reveals the critical role of the non-coding RNAs, particularly microRNAs (miRNAs), as vital regulatory elements in cancer.MethodWe performed a miRNAs microarray using paired tissues (tumor and adjacent normal bladder tissue), followed by the validation with qRT-PCR of five selected transcripts. Additional next-generation sequencing investigation established the interconnection among the altered miRNAs and mutated genes. Based on the overlapping between TCGA data and data obtained in the study, we focused on the systematic identification of altered miRNAs and genes mutated involved in bladder cancer tumorigenesis and progression.ResultsBy overlapping the miRNAs expression data, the two patient cohorts, we identified 18 miRNAs downregulated and, 187 miRNAs upregulated. qRT-PCR validation was completed using a selected panel of two downregulated (miR-139-5p and miR-143-5p) and three up-regulated miRNAs (miR-141b, miR-200 s or miR-205). Altered miRNAs patterns are interrelated to bladder tumorigenesis, allowing them to be used for the development of novel diagnostic and prognostic biomarkers. Three EMT-related upregulated miRNAs have an essential role in the molecular mechanisms, specifically key processes underlying tumorigenesis, invasion and metastasis. Using the Ampliseq Cancer Panel kit and Ion Torrent PGM Next-Generation Sequencing an increased mutation rate for TP53, FGFR3, KDR, PIK3CA and ATM were observed, but the mutational status for only TP53 was correlated to the survival rate. The miRNAs pattern, along with the gene mutation pattern attained, can assist for better patient diagnosis.ConclusionThis study thereby incorporates miRNAs as critical players in bladder cancer prognosis, where their altered gene expression profiles have a critical biological function in relationship with tumor molecular phenotype. The miRNA-mRNA regulatory networks identified in BC are ripe for exploitation as biomarkers or targeted therapeutic strategies.

Abstract 5247: Overexpression of methylenetetrahydrofolate dehydrogenase 1 like (MTHFD1L) in bladder cancer: A potential therapeutic target

Abstract Introduction: Methylenetetrahydrofolate dehydrogenase 1 (MTHFD1L) is a mitochondrial enzyme involved in the folate cycle by catalyzing the reaction of formyl-tetrahydrofolate to format and tetrahydrofolate (THF). THF is crucial for de novo purine and thymidylate synthesis and is also involved in the regeneration of methionine. Cancer cells rely on purines derived from the de novo pathway for the nucleotides whereas normal cells favor the salvage pathway. Thus, MTHFD1L could be a valuable therapeutic target for anticancer drug design. This study aimed to examine MTHFD1L expression in bladder cancer (BLCA). Material and Methods: Applying the publicly available cancer transcriptome database UALCAN, we searched for overexpressed genes in bladder cancer associated with survival probability. These in silico analyses indicated a high expression of MTHFD1L in bladder cancer. Furthermore, MTHFD1L expression was analyzed in different molecular subtypes, stage and race groups. Survival probability in MTHFD1L low-and high-expressing tumors was also assessed using UALCAN. Additionally, by searching the publicly available data base Human Protein Atlas we analyzed MTHFD1L immunostaining patterns in bladder cancer. Results: We analyzed data for 19 normal bladder tissues and 408 primary bladder cancers from the TCGA dataset. MTHFD1L was significantly (p<0.0001) overexpressed in tumorous tissue compared to normal. This overexpression was observed independent of stage, race and age. Furthermore, we found a higher number of MTHFD1L mRNA transcripts in tumors belonging to the basal/squamous and in the neuronal TCGA subtype. Kaplan-Meier survival analysis indicated a poorer survival in patients with high MTHFD1L expression (p=0.05). Evaluation of immunohistochemistry staining pattern in bladder cancer using the Human Protein Atlas database showed a differential expression among human bladder cancer tissues. We are performing in vitro studies to evaluate the role of MTHFD1L in bladder cancer initiation and progression. Conclusions: Our preliminary findings on MTHFD1L suggest that its overexpression is associated with poor patient survival in bladder cancer. These results suggest that MTHFD1L could be important in the biology of BLCA and a potential target for therapy. Future studies will investigate the functional significance of MTHFD1L and its regulation in bladder cancer. Citation Format: Marie-Lisa Eich, Maria D. Rodriguez Pena, Darshan S. Chandrashekar, Jennifer B. Gordetsky, George J. Netto, Sooryanarayana Varambally. Overexpression of methylenetetrahydrofolate dehydrogenase 1 like (MTHFD1L) in bladder cancer: A potential therapeutic target [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5247.

Evaluation of prognostic effects of MSI, PD-L1 and TIL in resected bladder cancer.

465 Background: The aim of the study was evaluating prognostic significance of tumor infiltrating lymphocytes (TIL), microsatellite instability (MSI) and programmed cell death ligand-1 (PD-L1) expression which are used as predictive biomarkers in immunotherapy. Methods: Clinic and pathologic features of bladder cancer patients who were underwent radical cystectomy were retrospectively analyzed from patients’ records in this single-center study. PD-L1, PD-L1 in TIL, PMS2, MSH2, MSH6 and MLH1 immunohistochemistry staining were carried out to archived resected tumor specimens of the eligible patients. MSI was evaluated according to existing of PMS2, MSH2, MSH6 and MLH1. Results: Fifty-six males (91.8%) and 5 females (8.2%) of total 61 patients were eligible for the study. Median age was 61.72 (35-81). PD-L1 expression in tumor cells and PD-L1 expression in TIL were positive in 14,8% (n = 9) and 16.4% (n = 10) of total 74 patients, respectively. MSI was high in 15 patients (24.6%). Intratumoral TIL rate was < 10% in 49 patients (80.3%) while > 10% in 12 patients. (19.7%) In Chi-square test, there was no statistically significant relationship between PD-L1, PD-L1 in TIL, MSI and TIL rate and patients’ characteristics including sex, stage, pathologic grade and lymph node status. Three-years overall survival (OS) rate was calculated as 43%. Univariate analysis did not reveal any significant difference in 3-years OS in PD-L1 expression in tumor, PD-L1 expression in TIL and TIL rate > 10% . (p = 0.822, p = 0.638, p = 0.318; respectively ). There was a positive trend with MSI high patients and OS (p = 0.089). At also; age and stage showed statistically significant difference in multivariate analysis. (p = 0.023, p = 0.011, respectively). Conclusions: This study revealed that there is no prognostic significance of PD-L1 and TIL. However, there are positive trend between OS and MSI, which is proven as predictive biomarker of immunotherapy, in patients who were followed with same treatment and follow-up approach in single center. These results must be corroborated with large and prospective investigations in order to better understanding biology and behavior of bladder cancer.

Epigenetic Mechanisms Influencing Epithelial to Mesenchymal Transition in Bladder Cancer.

Bladder cancer is one of the most incident neoplasms worldwide, and its treatment remains a significant challenge, since the mechanisms underlying disease progression are still poorly understood. The epithelial to mesenchymal transition (EMT) has been proven to play an important role in the tumorigenic process, particularly in cancer cell invasiveness and metastatic potential. Several studies have reported the importance of epigenetic mechanisms and enzymes, which orchestrate them in several features of cancer cells and, specifically, in EMT. In this paper, we discuss the epigenetic enzymes, protein-coding and non-coding genes, and mechanisms altered in the EMT process occurring in bladder cancer cells, as well as its implications, which allows for improved understanding of bladder cancer biology and for the development of novel targeted therapies.

Transcriptomic and Protein Analysis of Small-cell Bladder Cancer (SCBC) Identifies Prognostic Biomarkers and DLL3 as a Relevant Therapeutic Target.

Transcriptomic profiling can shed light on the biology of small-cell bladder cancer (SCBC), nominating biomarkers, and novel therapeutic targets. Sixty-three patients with SCBC had small-cell histology confirmed and quantified by a genitourinary pathologist. Gene expression profiling was performed for 39 primary tumor samples, 1 metastatic sample, and 6 adjacent normal urothelium samples (46 total) from the same cohort. Protein levels of differentially expressed therapeutic targets, DLL3 and PDL1, and also CD56 and ASCL1, were confirmed by IHC. A SCBC PDX model was utilized to assess in vivo efficacy of DLL3-targeting antibody-drug conjugate (ADC). Unsupervised hierarchical clustering of 46 samples produced 4 clusters that correlated with clinical phenotypes. Patients whose tumors had the most "normal-like" pattern of gene expression had longer overall survival (OS) compared with the other 3 clusters while patients with the most "metastasis-like" pattern had the shortest OS (P = 0.047). Expression of DLL3, PDL1, ASCL1, and CD56 was confirmed by IHC in 68%, 30%, 52%, and 81% of tissue samples, respectively. In a multivariate analysis, DLL3 protein expression on >10% and CD56 expression on >30% of tumor cells were both prognostic of shorter OS (P = 0.03 each). A DLL3-targeting ADC showed durable antitumor efficacy in a SCBC PDX model. Gene expression patterns in SCBC are associated with distinct clinical phenotypes ranging from more indolent to aggressive disease. Overexpression of DLL3 mRNA and protein is common in SCBC and correlates with shorter OS. A DLL3-targeted ADC demonstrated in vivo efficacy superior to chemotherapy in a PDX model of SCBC.

Cancer stem cells as a therapeutic target in bladder cancer.

Bladder cancer is one of the most prevalent genitourinary cancers responsible for about 150,000 deaths per year worldwide. Currently, several treatments, such as endoscopic and open surgery, appended by local or systemic immunotherapy, chemotherapy, and radiotherapy are used to treat this malignancy. However, the differences in treatment outcome among patients suffering from bladder cancer are considered as one of the important challenges. In recent years, cancer stem cells, representing a population of undifferentiated cells with stem-cell like properties, have been eyed as a major culprit for the high recurrence rate in superficial papillary bladder cancer. Cancer stem cells have been reported to be resistant to conventional treatments, such as chemotherapy, radiation, and immunotherapy, which induce selective pressure on tumoral populations resulting in selection and growth of the resistant cells. Therefore, targeting the therapeutic aspects of cancer stem cells in bladder cancer may be promising. In this study, we briefly discuss the biology of bladder cancer and then address the possible relationship between molecular biology of bladder cancer andcancer stem cells. Subsequently, the mechanisms of resistance applied by cancer stem cells against the conventional therapeutic tools, especially chemotherapy, are discussed. Moreover, by emphasizing the biomarkers described for cancer stem cells in bladder cancer, we have provided, described, and proposed targets on cancer stem cells for therapeutic interventions and, finally, reviewed some immunotargeting strategies against bladder cancer stem cells.

Molecular markers in bladder cancer

PurposeUse of molecular markers in urine, tissue or blood offers potential opportunities to improve understanding of bladder cancer biology which may help identify disease earlier, risk stratify patients, improve prediction of outcomes or help target therapy.MethodsA review of the published literature was performed, without restriction of time.ResultsDespite the fast-growing literature about the topic and the approval of several urinary biomarkers for use in clinical practice, they have not reached the level of evidence for widespread utilization. Biomarkers could be used in different clinical scenarios, mainly to overcome the limitations of current diagnostic, predictive, and prognostic tools. They have been evaluated to detect bladder cancer in asymptomatic populations or those with hematuria and in surveillance of disease as adjuncts to cystoscopy. There is also a potential role as prognosticators of disease recurrence, progression and survival both in patients with non-invasive cancers and in those with advanced disease. Finally, they promise to be helpful in predicting the response to local and/or systemic chemotherapy and/or immunotherapy.ConclusionsTo date, due to the lack of high-quality prospective trials, the level of evidence provided by the current literature remains low and, therefore, the potential of biomarkers exceeds utilization in clinical practice.

Molecular Subtype-Specific Immunocompetent Models of High-Grade Urothelial Carcinoma Reveal Differential Neoantigen Expression and Response to Immunotherapy.

High-grade urothelial cancer contains intrinsic molecular subtypes that exhibit differences in underlying tumor biology and can be divided into luminal-like and basal-like subtypes. We describe here the first subtype-specific murine models of bladder cancer and show that Upk3a-CreERT2; Trp53L/L; PtenL/L; Rosa26LSL-Luc (UPPL, luminal-like) and BBN (basal-like) tumors are more faithful to human bladder cancer than the widely used MB49 cells. Following engraftment into immunocompetent C57BL/6 mice, BBN tumors were more responsive to PD-1 inhibition than UPPL tumors. Responding tumors within the BBN model showed differences in immune microenvironment composition, including increased ratios of CD8+:CD4+ and memory:regulatory T cells. Finally, we predicted and confirmed immunogenicity of tumor neoantigens in each model. These UPPL and BBN models will be a valuable resource for future studies examining bladder cancer biology and immunotherapy.Significance: This work establishes human-relevant mouse models of bladder cancer. Cancer Res; 78(14); 3954-68. ©2018 AACR.

Bladder cancer rapid autopsy: A mechanism for collecting metastatic tumor tissue and establishing xenograft models of metastatic disease.

478 Background: Patients with metastatic bladder cancer have a poor prognosis with a median survival of only 9-15 months. Understanding the biology of metastatic bladder cancer has been historically difficult due to a paucity of specimens and models that recapitulate human disease. We established a bladder cancer rapid autopsy program to systemically acquire metastases, and build patient-derived xenograft (PDX) and organoid models for biological studies. Methods: Patients with metastatic bladder cancer were consented and a rapid autopsy was performed 2-6 hours after death to allow acquisition of normal and metastasis specimens. Frozen and formalin-fixed and paraffin embedded specimens were collected and pathological evaluation was performed on each specimen. Additionally, tumors were implanted subcutaneously into SCID mice to establish PDXs. Once PDXs were developed, PDXs were dissociated for companion organoid culture. Clinical history and treatment information was documented for each patient. Results: We have performed 10 bladder cancer rapid autopsies to date, and have acquired 105 metastatic and 45 normal specimens. The pathological subtypes of patients include urothelial cell carcinoma (5/10), squamous cell carcinoma (3/10), and plasmacytoid variant of urothelial carcinoma (2/10). The cohort has been treated BCG only (1/10), chemotherapy (3/10), and chemotherapy and immune checkpoint inhibitors (6/10). The leading site of metastasis was the liver (7/10) and lung (7/10), followed by lymph node (5/10), bone (5/10), intestine (4/10), and omentum (2/10). Half (5/10) of the patients had extensive liver metastases that allow acquisition of multiple tumor foci. For the first time, PDX and organoids from two independent metastases (omentum – CoCaB 14.1 and liver –CoCaB 14.2) from the same patient were successfully developed. Conclusions: This bladder cancer rapid autopsy program provides an important volume of metastatic tumor specimens for study. Importantly, the first bladder cancer PDX derived from metastasis has been developed. The availability of metastatic bladder cancer specimens, PDXs, and organoids will allow biological studies of advanced disease.

Oxidative damage and response to Bacillus Calmette-Gu\xe9rin in bladder cancer cells expressing sialyltransferase ST3GAL1

BackgroundTreatment with Bacillus Calmette-Guérin (BCG) is the gold standard adjuvant immunotherapy of non-muscle invasive bladder cancer (NMIBC), although it fails in one third of the patients. NMIBC expresses two tumor-associated O-linked carbohydrates: the disaccharide (Galβ1,3GalNAc) Thomsen-Friedenreich (T) antigen, and its sialylated counterpart (Siaα2,3Galβ1,3GalNAc) sialyl-T (sT), synthesized by sialyltransferase ST3GAL1, whose roles in BCG response are unknown.MethodsThe human bladder cancer (BC) cell line HT1376 strongly expressing the T antigen, was retrovirally transduced with the ST3GAL1 cDNA or with an empty vector, yielding the cell lines HT1376sT and HT1376T, that express, respectively, either the sT or the T antigens. Cells were in vitro challenged with BCG. Whole gene expression was studied by microarray technology, cytokine secretion was measured by multiplex immune-beads assay. Human macrophages derived from blood monocytes were challenged with the secretome of BCG-challenged BC cells.ResultsThe secretome from BCG-challenged HT1376sT cells induced a stronger macrophage secretion of IL-6, IL-1β, TNFα and IL-10 than that of HT1376T cells. Transcriptomic analysis revealed that ST3GAL1 overexpression and T/sT replacement modulated hundreds of genes. Several genes preserving genomic stability were down-regulated in HT1376sT cells which, as a consequence, displayed increased sensitivity to oxidative damage. After BCG challenge, the transcriptome of HT1376sT cells showed higher susceptibility to BCG modulation than that of HT1376T cells.ConclusionsHigh ST3GAL1 expression and T/sT replacement in BCG challenged-BC cancer cells induce a stronger macrophage response and alter the gene expression towards genomic instability, indicating a potential impact on BC biology and patient’s response to BCG.

A prospective multicenter study on bladder cancer: the COBLAnCE cohort.

BackgroundBladder cancer is a very heterogeneous disease as regards natural history. Environmental exposures, constitutional genetic and/or epigenetic background may affect not only the likelihood of bladder tumor occurrence, but also the histologic type of cancer and its outcome. Currently, only a few data are available to study the prognostic role of genetic and environmental factors. Likewise, data on the economic burden of bladder cancer and the longitudinal impact of the disease and the treatments on patient quality of life are scarce.MethodsCOBLAnCE is a large French-based clinical cohort study on bladder cancer. Newly diagnosed patients are enrolled prospectively in 12 public hospitals and 5 private for profits hospitals. The target sample size is 2,000 patients. All patients are to be followed for 6 years. Information on patient characteristics and lifestyle is collected during a face-to-face interview at enrollment. Clinical information on disease presentation, diagnosis, and treatment is extracted from medical records for the primary tumor and for all subsequent local and distant recurrences. Quality of life and resource use is collected at recruitment and during follow-up. In parallel, 4 types of biological samples (blood, tumor tissue, urine and nail) are collected, at baseline and during follow-up. DNA, RNA and PBMLs are extracted from blood samples, DNA and RNA from stabilized urine, proteins from frozen urine, DNA, RNA and proteins from frozen tumor tissues, and DNA and RNA from formalin-fixed paraffin-embedded tumor tissues. All derived products are stored at −80 °C or in liquid nitrogen. Main endpoints are gene-environment interactions, molecular classification, biomarker discovery, therapeutic innovation, treatment patterns, healthcare resource use, bladder cancer outcomes and quality of life.DiscussionThe COBLAnCE cohort will provide considerable insight into the biology of bladder cancer and the mechanisms through which genetic and environmental factors may influence the prognosis. It may allow the discovery of emerging biomarkers. Finally, economic data will be useful for future cost-effectiveness studies of immunotherapy drugs or other therapeutics in bladder cancer.

Protein Profiling of Bladder Urothelial Cell Carcinoma.

This study aimed to detect protein changes that can assist to understand the underlying biology of bladder cancer. The data showed forty five proteins were found to be differentially expressed comparing tumors vs non-tumor tissues, of which EGFR and cdc2p34 were correlated with muscle invasion and histological grade. Ten proteins (ß-catenin, HSP70, autotaxin, Notch4, PSTPIP1, DPYD, ODC, cyclinB1, calretinin and EPO) were able to classify muscle invasive BCa (MIBC) into 2 distinct groups, with group 2 associated with poorer survival. Finally, 3 proteins (P2X7, cdc25B and TFIIH p89) were independent factors for favorable overall survival.

Abstract 128: Comprehensive molecular characterization of 412 muscle-invasive urothelial bladder carcinomas: final analysis of The Cancer Genome Atlas (TCGA) project

Abstract Introduction: In 2014, TCGA's Bladder Cancer Working Group presented a preliminary integrated molecular analysis of 131 muscle-invasive urothelial carcinomas (Nature 507:315, 2014). We now report on the entire cohort of 412 fresh-frozen, chemotherapy-naïve tumors. Included in the analysis were paired blood and/or tumor-adjacent tissue samples. This is the largest sequencing project on bladder cancer to date. After strict clinical and pathologic quality control, tumors were analyzed for DNA copy number variants, somatic mutations, DNA methylation, mRNA, microRNA and (phospho-) protein expression, transcript splicing, gene fusions, viral integration, APOBEC mutagenesis, pathway perturbation, clinical correlates, and histopathology. Results: There was a high overall somatic mutation rate (8.0/Mb), with a median of 245 and mean of 348 coding-region mutations per sample. That is the third highest mutation rate among the cancer types profiled by TCGA (after cutaneous melanoma and non-small cell lung cancers). We identified 54 genes as significantly mutated, compared with 32 in the original report on 131 tumors. TP53 mutations were the most common (49%), and also quite common were mutations in a number of chromatin-modifying genes, including MLL2 (29%), KDM6A (26%), ARID1A (25%), MLL3 (19%), EP300 (15%), CREBBP (12%), and MLL (11%). Other cancer-related genes showing frequent mutations included PIK3CA (22%), RB1 (17%), FGFR3 (14%), STAG2 (14%), ATM (14%), ELF3 (12%), FAT1 (12%), SPTAN1 (12%), ERBB2 (12%), ERBB3 (11%), ASXL2 (10%), ERCC2 (9%), CDKN1A (9%), TSC1 (8%), CDKN2A (7%), RHOB (6%), NFE2L2 (6%), PARD3 (6%), FAM47C (5%), RBM10 (5%),HRAS (5%), KRAS (4%), and PTEN (3%). High mutation burden was associated with improved outcome (p = 0.0004). APOBEC mutagenesis explained 70% of the mutation burden and was associated with survival. Gene silencing by promoter hypermethylation was identified in 167 genes with at least 5% frequency in the cohort. The previously identified four mRNA expression subtypes were again found in the complete set of 412 tumors, and the proportions of samples in each subtype were similar to the previous proportions. Reverse-phase proteomic array analysis of 344 of the samples revealed clusters associated with diagnostic subtype, pathological stage, and grade but not with smoking history or non-muscle invasive status. Conclusions: This integrated molecular analysis of 412 TCGA tumor samples largely validates and considerably extends observations from the initial cohort of 131 patients. The larger cohort significantly increased our power to detect lower-frequency aberrations that were not identified in the original cohort. The results provide a robust basis for further functional studies of bladder cancer biology and also provide additional incisive information for the identification of molecular targets for therapy. Citation Format: John N. Weinstein, Seth P. Lerner, David J. Kwiatkowski, Gad Getz, Jaegil Kim, Hikmat A. Al-ahmadie, Andrew D. Cherniack, Guangwu Guo, Rehan Akbani, Katherine A. Hoadley, William Y. Kim, Gordon Robertson, Andrew J. Mungall, Toshinori Hinoue, Peter W. Laird, Jonathan E. Rosenberg, Joaquim Bellmunt, Dean F. Bajorin, Margaret B. Morgan, Chad J. Creighton, Dmitry Gordenin, Joshua M. Stuart, Xiaoping Su, Michael C. Ryan, Jeffrey S. Damrauer, Wei Zhang, Yuexin Liu, Yiling Lu, Nikolaus Schultz, Raju Kucherlapati, Gordon B. Mills, Donna E. Hansel, Brian D. Robinson, Bodgen A. Czerniak, Victor E. Reuter. Comprehensive molecular characterization of 412 muscle-invasive urothelial bladder carcinomas: final analysis of The Cancer Genome Atlas (TCGA) project. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 128.

Circulating cell-free DNA profiling of patients with advanced urothelial carcinoma

Circulating cell-free DNA profiling of patients with advanced urothelial carcinoma

Natural biology and management of nonmuscle invasive bladder cancer.

This article reviews the natural biology of noninvasive bladder cancer and its management strategies while summarizing the most recent advances in the field. Nonmuscle invasive bladder cancer (NMIBC) has a tendency to recur and progress. Risk stratification has helped triage patients but improved tools, including biomarkers, are still needed. Enhanced endoscopy with photodynamic imaging, narrow band imaging, optical coherence tomography and confocal laser endomicroscopy show promise for diagnosis, risk stratification and disease monitoring. Attempts at better treatment, especially in refractory high-risk cases, include the addition of intravesical hyperthermia, combination and sequential therapy with existing agents and the use of novel agents such as mycobacterial cell wall extract. New data are emerging regarding the potential role of active surveillance in low-risk patients. NMIBC represents a variety of disease states and continues to pose management challenges. As our understanding of tumor biology improves and technology advances, achieving better outcomes through individualized care may be possible.

Comprehensive characterization of 412 muscle invasive urothelial carcinomas: Final analysis of The Cancer Genome Atlas (TCGA) project.

405 Background: We reported the integrated molecular analysis of 131 tumors in 2014 (Nature 507:315, 2014) and now report on the entire cohort of 412 tumors from the TCGA project in chemotherapy-naïve muscle invasive urothelial bladder cancer. Methods: Following strict clinical and pathologic quality control, tumors were analyzed for DNA copy number variants, somatic mutations (WES), DNA methylation, mRNA, microRNA and (phospho-) protein expression, transcript splicing, gene fusions, viral integration, pathway perturbation, clinical correlates, and histopathology Results: There was a high overall somatic mutation rate (8.0/Mb), as previously reported, with a median of 245 and mean of 348 coding region mutations per sample. There were 54 significantly mutated genes (SMGs) (MutSig_2CV), increased from 32 in the original report. TP53 remained the most commonly mutated gene, and chromatin-modifying genes (MLL2, ARID1A, KDM6A) were also frequently mutated. KRAS, ERBB2, RB1, and ELF3 showed significant increased frequencies of mutation. High mutation burden was associated with improved outcome (p = 0.0004). APOBEC mutagenesis explained 70% of the mutation burden and also was associated with survival. 167 genes were silenced by promoter DNA hypermethylation in at least 5% of tumors. Several SMGs were also epigenetically silenced at various frequencies (ZNF773, CDKN2A, FAT1, CASP8). The previously identified four mRNA subtypes were predicted on the larger set identifying a similar proportion of samples in each subtype. We identified 5 expression subtypes based on reverse-phase protein arrays(344 tumors). Five miR expression subtypes were strongly associated with 4 mRNA-based subtypes (P = 3.8E-37) and associated with overall survival (P = 0.05). Conclusions: This integrated analysis of 412 TCGA patient samples validates and extends observations from the first 131 patients and significantly increases our power to detect additional low-frequency aberrations. The results provide a robust basis for functional studies to further our understanding of the biology of bladder cancer, and aid in the development of more precisely targeted therapies.

Abstract 4783: Novel molecular markers of bladder cancer progression identified by global chromatin profiling

Abstract Bladder cancer (BCa) is the 5th most prevalent cancer in the US, and the metastatic disease has very poor prognosis with 150,000 deaths per year worldwide. The use of molecular markers to predict disease progression has been largely unsuccessful, with only few therapeutically approachable mutations. Over 2,300 alterations in the coding sequences have been described in high-grade bladder cancer, encompassing frequent alterations in genes involving chromatin modifiers. Thus, chromatin organization may play a critical role in this tumor type. The development of tumor progression models from transitional carcinoma samples that are poorly tumorigenic and non-metastatic to highly metastatic to the lung or liver by serial in vivo passaging allows analysis of the mechanisms associated with tumor progression. We examined whole exome sequencing to identify mutations, gene expression, and DNase I hypersensitivity followed by deep sequencing to identify sites of chromatin modification during tumor progression. Bioinformatic analysis of was performed using algorithms developed at LRBGE. Comparative analysis combining multiple data sets was performed using Ingenuity Pathways. Mutations in several previously identified genes associated with bladder cancer: TP53, RAS and TERT, and mutations/deletions in several chromatin modifiers KDM6A and MLL2/3 were found in parental cells. Only few additional mutations, none identifiable as cancer driver mutations marked the progression to metastatic phenotype. Changes in gene expression were already evident in progression to tumorigenic state, with high score in cells in cell-to-cell interaction pathway marked by a decrease in cell adhesion, increase in genes associated with EMT and genes associated with cancer. Changes in global chromatin landscape occurred at many known genes correlated with pathways detected by expression analysis, but many were in novel sites distant to promoters, suggestive of novel distant enhancer and confirm massive reprogramming of regulatory networks. Many of these sites were unique for each stage of tumor progression and have potential clinical application. Specific chromatin modifications were found in NFkB and inflammatory pathways. Thus, in the tumorigenic stage, these cells acquired not only the capacity for tumor formation, but also some of the characteristics associated with metastasis. Combination of exome sequencing with the gene expression and unbiased analysis of global chromatin landscape provided valuable new information on bladder cancer biology and tumor progression. Citation Format: Lyuba Varticovski, Sohyoung Kim, Michael L. Nickerson, Bethtrice Thompson, Qizong Lao, Lars Grøntved, Songjoon Baek, Myong-Hee Sung, Dan Theodorescu, Michael Dean, Gordon L. Hager. Novel molecular markers of bladder cancer progression identified by global chromatin profiling. [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 4783. doi:10.1158/1538-7445.AM2015-4783

Abstract 548: Exosome as biomarkers and diagnostics in bladder cancer

Abstract Esosomes are small membrane vesicles secreted by most cell types. They are first found in reticulocytes and play important roles in cell-to-cell communication. Recent studies suggest that exosomes secreted by tumor cells containing tumor specific proteins and miRNAs. Previous study shows that exosomes purified from high grade bladder cancer promote cancer progression. This study aims to characterize the genetic contents in bladder cancer exosomes through mass spectrometry and miRNA array. Exosomes were purified from bladder cancers, immortalized bladder cells, and urine of healthy volunteers and muscle invasive bladder cancer patients. Via proteomics analysis, we have identified several cancer associated exosomal proteins that were found only in bladder cancer, including EDIL-3, an anagiogenic protein in our previous report. Functional characterization of those proteins is currently undertaken with a focus on periostin an extracellulear matrix protein. In addition to protein, exosomes also contain miRNAs that regulate cellular behaviors. Thus, we performed miRNA expression profiling microarray on exosomes from high grade bladder cancer cells, and immortalized bladder cells (LC Sciences). The differences in miRNA expression were compared and differences were determined by a student T test (P<0.05). Results showed significant difference miRNAs between cancer cells vs normal cells. Some of cancer-specific miRNAs which have shown to play critical roles in tumorigenesis and cancer progression will be study priority. Ultimately our work should contribute to the understanding of how exosomes contribute to bladder cancer biology and may elucidate novel urine exosomes markers for bladder cancer diagnosis. Note: This abstract was not presented at the meeting. Citation Format: Yu-Ru Liu, Christopher Silver, Yi-Fen Lee. Exosome as biomarkers and diagnostics in bladder cancer. [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 548. doi:10.1158/1538-7445.AM2015-548

MP68-16 ASSOCIATION OF GENETIC VARIANTS WITH THE OCCURRENCE OF BLADDER CANCER IN AN ARSENIC-EXPOSED POPULATION

You have accessJournal of UrologyBladder Cancer: Basic Research IV1 Apr 2015MP68-16 ASSOCIATION OF GENETIC VARIANTS WITH THE OCCURRENCE OF BLADDER CANCER IN AN ARSENIC-EXPOSED POPULATION Mario I. Fernandez, Cecilia Vial, Patricio Valdebenito, Eduardo Chaparro, Karena Espinoza, and Gabriela Repetto Mario I. FernandezMario I. Fernandez More articles by this author , Cecilia VialCecilia Vial More articles by this author , Patricio ValdebenitoPatricio Valdebenito More articles by this author , Eduardo ChaparroEduardo Chaparro More articles by this author , Karena EspinozaKarena Espinoza More articles by this author , and Gabriela RepettoGabriela Repetto More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2015.02.2478AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Inhabitants of Antofagasta (Northern Chile) were exposed to significantly elevated arsenic in drinking water between 1958 and 1971. As a consequence, incidence of bladder cancer (BC) in this city is currently 4 to 5 times higher than in the rest of the country (24.8 vs. 6/100,000). Carcinogenesis of arsenic-related BC is poorly understood to date. Therefore, we aimed to perform a genome-wide association study on individuals exposed to arsenic, looking for genetic variants associated to BC susceptibility among cases and controls. METHODS Subjects were invited to participate after signing an informed consent. Epidemiologic and clinical data (demographics, smoking history, family history of cancer and medical history) were collected during an in-person interview using a validated questionnaire and a blood sample was obtained. Patient variables were assessed by means of the Pearson χ2 test, comparing cases with controls. Differences in continuous variables were evaluated by the Student's t test. DNA samples were analyzed using Affymetrix Genome-Wide SNP Array 6.0. After filtering by missingness per individual, missingness per marker allele frequency and Hardy Weinberg Equilibrium we obtained 788,705 SNPs to be analyzed. RESULTS Males were predominant among cases and controls. Meanwhile, there were no significant differences in terms of mean age, familial history of BC, occupational exposure data or smoking status between groups. Tobacco smoking prevalence according to WHO classification was significantly frequent among cases and controls (59.5 and 58.3% respectively; p=0.55). The study population was found to be homogeneous after performance of principal component analysis for stratification, clustering the different individuals with the identity by state. Two regions with a significant association were identified after association tests comparing cases and controls: (a) rs4838646 in chromosome 10 (p=3.8E-06; OR 0.14) and (b) rs12371702 in chromosome 12 (p=5.8E-06; OR 7.2). Previous studies have linked polymorphisms in the former region to BC susceptibility. CONCLUSIONS Initial results of a BC genomic case-control study in an arsenic-exposed population are presented and raise candidate risk SNPs that need to be further validated in independent analyses. Additional insights into the genetics and biology of arsenic-related BC are also provided. However, further analysis is warranted after completion of recruitment. © 2015 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 193Issue 4SApril 2015Page: e863-e864 Advertisement Copyright & Permissions© 2015 by American Urological Association Education and Research, Inc.MetricsAuthor Information Mario I. Fernandez More articles by this author Cecilia Vial More articles by this author Patricio Valdebenito More articles by this author Eduardo Chaparro More articles by this author Karena Espinoza More articles by this author Gabriela Repetto More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...