Disease Progression In Bladder Cancer Research Articles

Abstract 3762: 11,000-plex aptamer based proteomics of bladder cancer urine for the discovery of novel disease biomarkers

Abstract In search of more accurate, non-invasive biomarkers of bladder cancer (BC), a comprehensive aptamer-based screen of urine proteins was conducted using urine samples from a BC cohort, followed by systems biology analyses. Following an initial 1317-plex screen, an extended 11,000 plex aptamer-based proteomic screen is in progress. Based on the initial screen, 30 urine proteins were next ELISA validated in an independent cohort of 68 subjects, and the results are shown in Table 1. Of 21 urine proteins that discriminated BC from urology clinic controls (UC), urine D-dimer displayed the highest ROC AUC value (0.96) and sensitivity (95%) (Table 1). Furthermore, 9 urine proteins significantly discriminated more advanced BC (T1-T4) from earlier stages (Ta-Tis), with IL-8 and MMP-12 being the best performers. Urine MMP12 exhibited the highest specificity of 80% at fixed sensitivity for identifying advanced BC. Systems biology analysis implicated molecular functions related to extracellular matrix, collagen, integrin, heparin and transmembrane tyrosine kinase signaling in BC susceptibility, with HNF4A and NFKB1 emerging as key molecular regulators. STEM analysis of the dysregulated pathways implicated a functional role for the immune system, complement and interleukins in BC disease progression. Urine C2, D-dimer, and Elastase were additionally ELISA validated in a second BC cohort of Chinese ethnicity. Thus, comprehensive proteomics and independent ELISA validation have uncovered novel urine biomarkers for initial BC diagnosis and subsequent disease progression, that warrant further head to head comparison with current diagnostic tests. Citation Format: Chandra Mohan, Kamala Vanarsa, Jessica Castillo, Long Wang, Yair Lotan. 11,000-plex aptamer based proteomics of bladder cancer urine for the discovery of novel disease biomarkers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3762.

Significance and Mechanisms Analyses of RB1 Mutation in Bladder Cancer Disease Progression and Drug Selection by Bioinformatics Analysis.

Bladder cancer is still a disease of significant morbidity and mortality. In bladder cancer, RB1 is one of the most common mutant genes. In this study, we explored the Genomics of Drug Sensitivity in Cancer (GDSC) database for drug sensitivity. The latest TCGA data were downloaded for analysis. To deal with functional enrichment analysis, GSEA, KEGG and GO were used. Prognostic analyses have been carried out using the GEPIA online tool. Results from the GDSC database showed that bladder cancer cells with RB1 mutation are more resistant to Dactolisib, MK-2206 and GNE-317. RB1 mutation was found in 25%bladder cancer patients. Patients with RB1 mutation often had lower RB1 mRNA expression level and higher histologic grade. In addition, we identified 999 differentially expressed genes in both groups. Functional enrichment analysis suggested that DEGs were primarily enriched in multiple metabolic progressions, cell proliferation and cancer related pathways. There were strong correlations between WT1, GPR37, CHRM2 and EZH2 expression levels and the prognosis. In all, the significance of RB1 mutation in disease progression and drug selection in bladder cancer was suggested by our results, and multiple genes and pathways related to such a program were identified.

Urinary glycoproteomic profiling of non-muscle invasive and muscle invasive bladder carcinoma patients reveals distinct N-glycosylation pattern of CD44, MGAM, and GINM1.

Clinical management of bladder carcinomas (BC) remains a major challenge and demands comprehensive multi-omics analysis for better stratification of the disease. Identification of patients on risk requires identification of signatures predicting prognosis risk of the patients. Understanding the molecular alterations associated with the disease onset and progression could improve the routinely used diagnostic and therapy procedures. In this study, we investigated the aberrant changes in N-glycosylation pattern of proteins associated with tumorigenesis as well as disease progression in bladder cancer. We integrated and compared global N-glycoproteomic and proteomic profile of urine samples from bladder cancer patients at different clinicopathological stages (non-muscle invasive and muscle-invasive patients [n = 5 and 4 in each cohort]) with healthy subjects (n = 5) using SPEG method. We identified 635 N-glycopeptides corresponding to 381 proteins and 543 N-glycopeptides corresponding to 326 proteins in NMIBC and MIBC patients respectively. Moreover, we identified altered glycosylation in 41 NMIBC and 21 MIBC proteins without any significant change in protein abundance levels. In concordance with the previously published bladder cancer cell line N-glycoproteomic data, we also observed dysregulated glycosylation in ECM related proteins. Further, we identified distinct N-glycosylation pattern of CD44, MGAM, and GINM1 between NMIBC and MIBC patients, which may be associated with disease progression in bladder cancer. These aberrant protein glycosylation events would provide a novel approach for bladder carcinoma diagnosis and further define novel mechanisms of tumor initiation and progression.

Significance of TP53 mutation in bladder cancer disease progression and drug selection.

BackgroundThe tumor protein p53 (TP53) mutant is one of the most frequent mutant genes in bladder cancer. In this study, we assessed the importance of the TP53 mutation in bladder cancer progression and drug selection, and identified potential pathways and core genes associated with the underlying mechanisms.MethodsGene expression data used in this study were downloaded from The Cancer Genome Atlas and cBioportal databases. Drug sensitivity data were obtained from the Genomics of Drug Sensitivity in Cancer. We did functional enrichment analysis by gene set enrichment analysis (GSEA) and the Database for Annotation, Visualization and Integrated Discovery (DAVID).ResultsWe found the TP53 mutation in 50% of bladder cancer patients. Patients with the TP53 mutation were associated with a lower TP53 mRNA expression level, more advanced tumor stage and higher histologic grade. Three drugs, mitomycin-C, doxorubicin and gemcitabine, were especially more sensitive to bladder cancer with the TP53 mutation. As for the mechanisms, we identified 863 differentially expressed genes (DEGs). Functional enrichment analysis suggested that DEGs were primarily enriched in multiple metabolic progressions, chemical carcinogenesis and cancer related pathways. The protein–protein interaction network identified the top 10 hub genes. Our results have suggested the significance of TP53 mutation in disease progression and drug selection in bladder cancer, and identified multiple genes and pathways related in such program, offering novel basis for bladder cancer individualized treatment.

Disease progression in bladder cancer: which developments since 1994

Objectives The introduction of PSA in clinical practice has resulted in decreasing the death rate form prostate cancer and in a downward shift of the pathological stage in radical prostatectomy specimens. This seems not to be the case for bladder cancer. In order to verify this assumption, we have reviewed the charts of the patients operated on of radical prostatectomy and radical cystectomy between 1994 and 2006. METHODS 456 and 491 consecutive patients, respectively, underwent radical cystectomy and radical prostatectomy with bilateral lymph nodes dissection. We excluded all the patients who had received neoadjuvant treatment or did not undergo node dissection. The patients were divided into two consecutive groups according to the year of treatment: group 1 included pts treated from 1994 to 2000, and group 2 pts from 2001 to 2006. The histopathological findings of the two groups of pts were compared. The difference among TNM systems has been balanced evaluating histopathological reports critically and converting them to the 2002 edition. RESULTS. For patients with prostate cancer, those in group 2 had a decrease in the incidence of extracapsular extension and lymph nodes invasion. The bladder cancer patients belonging to group 2 had a greater number of T2, but there was an increased number of pN+ in this group. CONCLUSIONS Even if there is a decline in locally advanced disease in patients with bladder cancer, our retrospective analysis did not show a comparable success in early diagnosis as it did for prostate cancer. There is undoubtedly an increase in the lymph node dissemination, whether this is due to a more extended lymph node dissection or to a premature dissemination remains questionable. Public awareness regarding bladder cancer and its risk factors is limited, but several studies have reported that a delay in diagnosis of invasive bladder cancer is an adverse prognostic factor. A higher care in the development of new diagnostic markers for bladder tumors and especially in the screening protocols together with an earlier radical therapy could hopefully improve the management of such a pathology, as it happened for prostate cancer.

Expression of the Long Non-Coding RNA HOTAIR Correlates with Disease Progression in Bladder Cancer and Is Contained in Bladder Cancer Patient Urinary Exosomes.

Exosomes are 30-150nM membrane-bound secreted vesicles that are readily isolated from biological fluids such as urine (UEs). Exosomes contain proteins, micro RNA (miRNA), messenger RNA (mRNA), and long non-coding RNA (lncRNA) from their cells of origin. Although miRNA, protein and lncRNA have been isolated from serum as potential biomarkers for benign and malignant disease, it is unknown if lncRNAs in UEs from urothelial bladder cancer (UBC) patients can serve as biomarkers. lncRNAs are > 200 nucleotide long transcripts that do not encode protein and play critical roles in tumor biology. As the number of recognized tumor-associated lncRNAs continues to increase, there is a parallel need to include lncRNAs into biomarker discovery and therapeutic target algorithms. The lncRNA HOX transcript antisense RNA (HOTAIR) has been shown to facilitate tumor initiation and progression and is associated with poor prognosis in several cancers. The importance of HOTAIR in cancer biology has sparked interest in using HOTAIR as a biomarker and potential therapeutic target. Here we show HOTAIR and several tumor-associated lncRNAs are enriched in UEs from UBC patients with high-grade muscle-invasive disease (HGMI pT2-pT4). Knockdown of HOTAIR in UBC cell lines reduces in vitro migration and invasion. Importantly, loss of HOTAIR expression in UBC cell lines alters expression of epithelial-to-mesenchyme transition (EMT) genes including SNAI1, TWIST1, ZEB1, ZO1, MMP1 LAMB3, and LAMC2. Finally, we used RNA-sequencing to identify four additional lncRNAs enriched in UBC patient UEs. These data, suggest that UE-derived lncRNA may potentially serve as biomarkers and therapeutic targets.

Cell-cycle control in urothelial carcinoma: large-scale tissue array analysis of tumor tissue from Maine and Vermont.

Cell-cycle proteins are important predictive markers in urothelial carcinoma but may also exhibit exposure-specific heterogeneity. Tumor tissue from 491 bladder cancer cases enrolled in the Maine and Vermont component of the New England Bladder Cancer Study was assembled as tissue microarrays and examined for aberrant expression of p53, p63, p16, cyclin D1, Rb, and Ki-67. The association between expression and histopathology, demographics, and cigarette smoking was examined using χ(2) tests, multivariable Poisson, and multinomial regression models. We found that overexpression of p53 and Ki-67 was associated with high-stage/grade tumors [relative risk (RR), 1.26; P(trend) = 0.003; and RR, 3.21; P(trend) < 0.0001, respectively], whereas expression of p63 and p16 was decreased in high-stage/grade tumors (RR, 0.52; P(trend) < 0.0001; and RR, 0.88; P(trend) = 0.04, respectively). No significant aberrations of cell-cycle proteins were identified using various smoking variables and multiple statistical models. The results of this population-based study of histologically confirmed urothelial carcinomas show significant aberration of cell-cycle proteins p53, p63, p16, and Ki-67, but not Rb or cyclin D1. p53 showed the most significant heterogeneity with respect to tumor stage and grade, especially when stratified for different staining intensities using novel digital image analysis techniques. Our findings do not support that smoking modifies expression of cell-cycle proteins. Our study shows significant heterogeneity in the expression of key cell-cycle proteins that are associated with disease progression in bladder cancer. Further studies may lead to the identification of biomarkers and their multiplexed interactions as useful prognostic and therapeutic targets.

Abstract 300: Identification of a panel of protein markers associated with disease progression in bladder cancer

Abstract Most bladder cancer patients are initially diagnosed with noninvasive or superficially invasive urothelial tumors, comprising benign low grade Ta tumors that rarely progress to high grade T1 tumors with concomitant CIS that progress in up to 60% of cases. So far, no biomarkers are available for solid prediction of the individual disease course. Previously, we identified an 88-transcript signature for prediction of progression for patients with non-muscle invasive bladder cancer. Aim: validation of key candidate markers on the protein level. We selected four protein coding genes, where high transcript levels were related to a reduced risk of progression and five genes where high expression identified patients with an increased risk of progression. Microarray transcript profiling data on 7 bladder cancer cell lines were used to select appropriate model cell lines. Specificity of antibodies against the selected proteins, protein expression and potential splice variants were investigated by western blotting using extracts from bladder cancer cell lines (T24, SW780) and HeLa cells. Specific antibodies were identified for six of the nine selected proteins. Immunohistochemistry was applied to a Tissue-microarray with 289 primary non-muscle invasive urothelial tumors (182 pTa, 101 pT1 and 6 CIS). 118 patients later experienced disease progression to pT2-4 bladder cancer (median time to progression 22 months). None of the remaining 171 patients showed later disease progression within the follow-up period (median follow-up 93 months). In univariate Cox regression analysis, expression of 4 of the 6 proteins correlated significantly with disease progression from stage Ta or T1 to muscle invasive bladder cancer. A reduced risk of progression correlated significantly with a strong nuclear expression of the G1/S-specific cell cycle protein (p=0.0003) and its expression in &amp;gt;80% nuclei (p=0.0004) as well as a membranous localization of a disintegrin and metallopeptidase (p = 0.0002). Increased risk of progression correlated significantly with strong nuclear and cytoplasmic expression of an ubiquitin-conjugating enzyme (p=0.0001) and its expression in &amp;gt;10% nuclei (p=0.0000) and with the expression of a DNA replication licensing factor in &amp;gt;80% nuclei (p=0.0026). In multivariate Cox regression analysis, the G1/S-specific cell cycle protein (HR=0.62, p=0.020, 95%CI 0.42-0.92), the disintegrin and metallopeptidase (HR=0.56, p=0.008, 95%CI 0.37-0.86) and the ubiquitin-conjugating enzyme (HR=1.68, p=0.015, 95%CI 1.1-2.5) were independent prognostic variables for predicting disease progression to muscle invasive bladder cancer when stratifying for other known clinical and histopathological risk factors as stage, age and grade. The Ubiquitin-conjugating enzyme was selected for in vitro functional analyses probably affecting cell proliferation and cell migration of bladder cancer cell lines. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 300. doi:10.1158/1538-7445.AM2011-300

Abstract 3438: Oncogenes-induced interferon-beta: modulation of tranformation phenotypes through interferon-ISG15 autocrine signaling

Abstract ISG15 (interferon-stimulated gene 15), an ubiquitin-like protein (UBL), is a new tumor marker whose expression has been correlated with disease progression in bladder cancer and suggested to be prognostic for breast cancer. However, the molecular basis for, and the function of, ISG15 overexpression in tumors remain unclear. Here, we show that ISG15 overexpression in both breast cancer cells and oncogenic Ras-transformed mammary epithelial cells (MCF-10A) is due to elevated secretion of interferon-β. Moreover, interferon-β-ISG15 autocrine signaling in oncogenic Ras-transformed MCF-10A cells results in increased colony formation efficiency. Interestingly, gain of epithelial-mesenchymal transition (EMT) characteristics, including appearance of spindle-shaped cell morphology and EMT markers, as well as increased cell migration, in Ras-transformed MCF-10A cells is also dependent on interferon-β-ISG15 autocrine signaling. Our results demonstrate for the first time that tumor cell-secreted interferon-β may modulate tumor cell growth and migration through ISG15, suggesting potential cancer therapeutic approaches by antagonizing interferon autocrine signaling. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3438. doi:10.1158/1538-7445.AM2011-3438

139 Annexin A10 (ANXA10) is a marker for metastasis and disease progression in bladder cancer

139 Annexin A10 (ANXA10) is a marker for metastasis and disease progression in bladder cancer

Disease Progression in Bladder Cancer: Which Developments since 1994

Objectives The introduction of PSA in clinical practice has resulted in decreasing the death rate form prostate cancer and in a downward shift of the pathological stage in radical prostatectomy specimens. This seems not to be the case for bladder cancer. In order to verify this assumption, we have reviewed the charts of the patients operated on of radical prostatectomy and radical cystectomy between 1994 and 2006. Methods 456 and 491 consecutive patients, respectively, underwent radical cystectomy and radical prostatectomy with bilateral lymph nodes dissection. We excluded all the patients who had received neoadjuvant treatment or did not undergo node dissection. The patients were divided into two consecutive groups according to the year of treatment: group 1 included pts treated from 1994 to 2000, and group 2 pts from 2001 to 2006. The histopathological findings of the two groups of pts were compared. The difference among TNM systems has been balanced evaluating histopathological reports critically and converting them to the 2002 edition. Results For patients with prostate cancer, those in group 2 had a decrease in the incidence of extracapsular extension and lymph nodes invasion. The bladder cancer patients belonging to group 2 had a greater number of T2, but there was an increased number of pN+ in this group. Conclusions Even if there is a decline in locally advanced disease in patients with bladder cancer, our retrospective analysis did not show a comparable success in early diagnosis as it did for prostate cancer. There is undoubtedly an increase in the lymph node dissemination, whether this is due to a more extended lymph node dissection or to a premature dissemination remains questionable. Public awareness regarding bladder cancer and its risk factors is limited, but several studies have reported that a delay in diagnosis of invasive bladder cancer is an adverse prognostic factor. A higher care in the development of new diagnostic markers for bladder tumors and especially in the screening protocols together with an earlier radical therapy could hopefully improve the management of such a pathology, as it happened for prostate cancer.

Gain of 5p15.33 Is Associated with Progression of Bladder Cancer

Objective: To search for a biological marker to distinguish low-risk from high-risk bladder cancer indicating disease progression. Methods: The whole genome-wide copy numbers were screened in 18 patients with bladder cancer using array comparative genomic hybridization (CGH) consisting of 4,030 bacterial artificial chromosome clones. Results: Gain of 5p15.33, including TPPP (tubulin polymerization-promoting protein)and ZDHHC11 (zinc finger DHHC domain-containing protein 11) genes, was detected in 5 of 9 (55.6%) high-grade bladder cancers and no (0%; n = 9) low-grade bladder cancer. To confirm the preliminary data, 5p15.33 gain was studied by fluorescence in situhybridization (FISH) in 100 patients, and the results were compared with biological characteristics. In FISH analysis, gain of 5p15.33 was significantly correlated with higher histological grade (p < 0.0001) and advanced pathological stage (p = 0.0284). Tumors with a gain of 5p15.33 had a significantly higher progression-free survival rate than those without (p = 0.0006, log-rank test). Multivariate analysis revealed that gain of 5p15.33 was a predictor for disease progression in bladder cancer (hazard ratio: 1.887, 95% confidence interval: 1.215–2.968, p = 0.0050). Conclusion: These data suggest that gain of 5p15.33 (TPPP and ZDHHC11) may become a potential biomarker identifying high-risk patients with disease progression in bladder cancer.

No association between HER-2 gene polymorphism at codon 655 and a risk of bladder cancer.

The amplification and overexpression of the human epidermal growth factor receptor 2 gene HER-2 (also known as c-erb-B2 or neu) have been shown to be associated with bladder cancer and its progression. Recent studies indicated an association between the Ile to Val polymorphism at codon 655 of HER-2 and susceptibility to breast cancer. To investigate the correlation between the Ile/Val polymorphism and the susceptibility and progression of bladder cancer, we analyzed the polymorphism in 232 patients with transitional cell carcinoma of the bladder and 408 normal controls. The frequencies of the Ile/Ile, Ile/Val and Val/Val genotype were 75.9%, 21.6% and 2.6%, respectively, in patients with bladder cancer and 75.7%, 23.0% and 1.2%, respectively, in controls. Statistical analyses of the genotype prevalence showed no significant difference between bladder cancer patients and normal controls (p = 0.419). Moreover, no significant differences in the genotype prevalence were observed when the patients were stratified according to the tumor grade, stage and smoking habits. When the Ile/Ile genotype was compared to the Ile/Val and Val/Val genotypes, a significant difference was found only between the patients with tumor stage Ta and those with T1-4 (age, gender and smoking habits-adjusted odds ratio = 2.13, 95% confidence interval = 1.09-4.15, p = 0.027). When the Ile/Ile + Ile/Val genotypes compared to the Val/Val genotype, no significant findings were observed. These results suggested that the HER-2 polymorphism at codon 655 is unlikely to be associated with the onset of bladder cancer. Furthermore, the findings suggest no association between this polymorphism and the disease progression in bladder cancer, although the possibility remains that the Ile/Ile genotype may be related to an increased risk of disease progression.