Survival In Muscle-invasive Bladder Cancer Patients Research Articles

Identification of a cisplatin resistant-based prognostic immune related gene signature in MIBC

Cisplatin resistance plays a significant role in the dismal prognosis and progression of muscle-invasive bladder cancer (MIBC). However, the strategies to predict prognosis and cisplatin resistance are inefficient, and it remains unclear whether cisplatin resistance is associated with tumor immunity. In this study, we integrated the transcriptional data from cisplatin-resistant cell lines and a TCGA-MIBC cohort to establish cisplatin-resistance-related cluster classification and a cisplatin-resistance-related gene risk score (CRRGRS). Kaplan–Meier survival curves showed that compared with those in low CRRGRS group, MIBC patients belonging to high CRRGRS group had worse prognosis in TCGA-MIBC cohort and external GEO cohorts. Meanwhile, CRRGRS was able to help forecast chemotherapy and immunotherapy response of MIBC patients in the TGCA cohort and IMvigor210 cohort. Moreover, compared with the low CRRGRS group, the high CRRGS group possessed a relatively immunosuppressive “cold tumor” phenotype with a higher tumor immune dysfunction and exclusion (TIDE) score, ESTIMATE score, stromal score and immune score and a lower immunophenoscore (IPS) score. The upregulated expression levels of immune checkpoint genes, including PD-1, PD-L1 and CTLA4, in the high CRRGRS group also further indicated that a relative immunosuppressive tumor microenvironment may exist in MIBC patients belonging to high CRRGRS group. In addition, we integrated CRRGRS and clinical characteristics with prognostic value to develop a nomogram, which could help forecast overall survival of MIBC patients. Furthermore, DIAPH3 was identified as a regulator of proliferation and cisplatin resistance in MIBC. The expression of DIAPH3 was increased in cisplatin-resistant cell lines and chemotherapy-unsensitive people. Further mechanism exploration revealed that DIAPH3 facilitated tumor proliferation and cisplatin resistance by regulating the NF-kB and epithelial-mesenchymal transition (EMT) pathways. In conclusion, the comprehensive investigations of CRRGRS increased the understanding of cisplatin resistance and provided promising insights to restrain tumor growth and overcome chemoresistance by targeting DIAPH3.

Role of Serum Lymphocyte-derived Biomarkers in Nonmetastatic Muscle-invasive Bladder Cancer Patients Treated with Trimodal Therapy

BackgroundThe role of serum lymphocyte-based biomarkers, such as the neutrophil-to-lymphocyte (NLR), lymphocyte-to-monocyte (LMR), and platelet-to-lymphocyte (PLR) ratios, was previously studied in patients with muscle-invasive bladder cancer (MIBC) treated with radical cystectomy but remains underexplored in patients treated with trimodal therapy (TMT). ObjectiveTo analyze the impact of serum lymphocyte-based biomarkers on main oncological outcomes after TMT for MIBC. Design, setting, and participantsA retrospective study, including 176 patients treated with TMT for nonmetastatic MIBC (cT2–4/cN0–2) between 2001 and 2017 at a tertiary academic center, was conducted. InterventionTMT, consisting of initial maximal transurethral resection of the bladder tumor, followed by radiotherapy with concurrent chemotherapy. Outcome measurements and statistical analysisClinicopathological characteristics, serum laboratory tests, and imaging reports were collected. NLR, LMR, and PLR were calculated before and at the end of TMT. Dynamic patterns of NLR, LMR, and PLR during TMT were studied. Multivariable regression models were performed to estimate the effect of these biomarkers on complete response (CR) to TMT and survival. Results and limitationsThe median age was 75 yr (interquartile range 66–82). Staging was cT2 in 156 (89%) and cN0 in 159 (90%) patients. A pretreatment NLR (pre-NLR) of ≥4.0 was independently associated with lower CR rates (odds ratio 0.32; p = 0.013). In addition, a pre-NLR of ≥4.0 was associated with worse cancer-specific survival (hazard ratio [HR] 1.88; p = 0.032) and overall survival (OS; HR 1.61; p = 0.033) together with other factors such as hydronephrosis, Eastern Cooperative Oncology Group performance status, and cT stage 3-4a. When both pre- and post-treatment variables were considered, an increase in NLR beyond 75% during TMT (HR 1.63; p = 0.035) was associated with worse OS. This study was limited by its retrospective design. ConclusionsA high pre-NLR value was independently associated with lower rates of CR and worse survival in MIBC patients undergoing TMT. Prospective validation is needed to implement NLR into clinical practice. Patient summaryIn this study, we reported the oncological outcomes of patients with muscle-invasive bladder cancer treated with trimodal therapy. We found that the neutrophil-to-lymphocyte ratio, a cheap and available blood-derived biomarker, was associated with response to trimodal therapy and survival outcomes.

A Competing Endogenous RNA Network and an 8-lncRNA Prognostic Signature Identify MYO16-AS1 as an Oncogenic lncRNA in Bladder Cancer

Recently, growing evidence has shed light on the competitive endogenous RNAs (ceRNAs) activity of long noncoding RNAs (lncRNAs) in carcinogenesis and tumor progression. To better elucidate the regulatory mechanisms of lncRNA in muscle-invasive bladder cancer (MIBC), we identified aberrantly expressed mRNAs, lncRNAs, and miRNAs in tumor tissues by using RNA sequence profiles from The Cancer Genome Atlas. The MIBC-specific ceRNA network, including 58 lncRNAs, 22 miRNAs, and 52 mRNAs, was constructed and visualized in Cytoscape. Further, using the univariate and multivariate Cox regression model, we screened 8 lncRNAs (AC078778.1, LINC00525, AC008676.1, AP000553.1, SACS-AS1, AC009065.1, AC127496.3, and MYO16-AS1) to construct an lncRNA signature for predicting the overall survival of MIBC patients. Kaplan-Meier analysis and a receiver operating characteristic curve were applied to evaluate the performance of the signature. Real-time quantitative PCR analysis was carried out to test expression levels of the 8 lncRNAs in MIBC patient tissues. Transwell assays demonstrated that overexpressing MYO16-AS1 can enhance UMUC2 migration and invasion. Our study offers a novel lncRNA-correlated ceRNA model to better understand the molecular mechanisms involved in MIBC. In addition, we developed an independent 8-lncRNAs biomarker for prognostic prediction and identified MYO16-AS1 as an oncogenic lncRNA in bladder cancer.

Meta-analysis of neoadjuvant chemotherapy compared to radical cystectomy alone in improving overall survival of muscle-invasive bladder cancer patients

BackgroundMost patients with muscle-invasive bladder cancer (MIBC) developed metastasis within 2 years, even after radical cystectomy (RC). The recurrence rate of MIBC was more than 50% of the cases. A meta-analysis conducted by Yin et al. showed that neoadjuvant chemotherapy (NAC) + RC improves overall survival in MIBC compared with RC only. However, a new meta-analysis by Li et al. concluded that NAC + RC was not superior to RC only in improving overall survival. The inconsistencies of these studies required further comprehensive analysis to recommend NAC use in bladder cancer treatment. Therefore, this meta-analysis aims to analyze previous studies that compare the efficacy of NAC + RC versus RC only to improve overall survival of MIBC.MethodsThe articles were searched using Pubmed with keywords “muscle-invasive bladder cancer”, “neoadjuvant chemotherapy”, “cystectomy”, and “overall survival”. The articles that were published until June 2020 were screened. The overall survival outcome was analyzed as hazard ratio (HR) and presented in a forest plot.ResultSeventeen studies were included in meta-analysis with a total sample of 13,391 patients, consist of 2890 received NAC followed by RC and 10,418 underwent RC only. Two studies used methotrexate/vinblastine/doxorubicin/cisplatin (MVAC), two studies used gemcitabine/cisplatin (GC), one study used Cisplatin-based regimen, one study used MVAC or GC, one study used gemcitabine/carboplatin (GCarbo) or GC or MVAC, one study used Cisplatin/Gemcitabine or MVAC, one study used Cisplatin only, one study used Cisplatin-based (GC, MVAC) or non-Cisplatin-based (combined paclitaxel/gemcitabine/carboplatin), one study used GC, MVAC, Carboplatin, or Gemcitabine/Nedaplatin (GN), and five studies did not mention the regimen The overall survival in the NAC + RC only group was significantly better than the RC only group (HR 0.82 [0.71–0.95], p = 0.009).ConclusionNAC + RC is recommended to improve overall survival in MIBC patients. A further study assessing side effects and quality of life regarding NAC + RC is needed to establish a strong recommendation regarding this therapy.

Abstract IA24: Predictive biomarkers of response to neoadjuvant therapy

Abstract The addition of cisplatin-based neoadjuvant chemotherapy prior to radical cystectomy provides a modest overall survival benefit for patients with muscle-invasive bladder cancer (MIBC). Moreover, the extent of pathologic response at cystectomy correlates with survival, and patients who achieve a complete pathologic response following neoadjuvant chemotherapy have significantly improved survival compared to patients with residual MIBC. Clinical features alone are unable to predict which patients will respond to neoadjuvant chemotherapy, and there is an urgent clinical need to identify genomic biomarkers to guide neoadjuvant therapy for MIBC. The Cancer Genome Atlas (TCGA) and related efforts have defined the genomic landscape of MIBC and have identified recurrent genomic alterations including mutations, chromosomal translocations, and copy number alterations. Additionally, gene expression profiling has defined molecular subtypes of MIBC that have distinct clinical and genomic features. These genomic data are being harnessed to understand the association among genomic features, response to neoadjuvant therapy, and clinical outcomes. Alterations in DNA damage and repair (DDR) genes have emerged as a biomarker of response to neoadjuvant cisplatin-based chemotherapy in several studies. Loss-of-function mutations in the nucleotide excision repair gene ERCC2 as well as in related DDR genes including ATM, FANCC, and RB1 have been associated with complete pathologic response to cisplatin-based chemotherapy and improved survival in MIBC patients receiving cisplatin-based chemotherapy. Functional studies have confirmed that many of the clinically identified DDR gene mutations confer loss of DNA repair capacity and drive cisplatin sensitivity in preclinical bladder systems. These data have informed the design of several ongoing biomarker-driven clinical trials that seek to identify a subset of patients with DDR-altered tumors who may be cured with cisplatin-based chemotherapy alone. In addition to driving response to DNA damaging-based chemotherapy, DDR gene alterations have also recently been associated with improved response to immune checkpoint inhibition in patients with metastatic urothelial cancer. The mechanism underlying this association remains incompletely understood; however, DDR pathway alterations can lead to increased tumor mutational burden and neoantigen load, which have each been associated with improved response to anti-PD1/PD-L1 agents. Immune checkpoint inhibitors have transformed the management of metastatic bladder cancer, and these agents are now being investigated in numerous clinical trials for patients with localized bladder cancer. However, only a subset of patients benefit from anti-PD1/PD-L1 therapy; therefore, identifying reliable predictive biomarkers of immunotherapy response in bladder cancer remains a critical need. Gene expression-based analyses have defined molecular subtypes that exhibit distinct clinical properties and therapeutic vulnerabilities. Tumors with basal-like gene expression properties appear to respond more robustly to neoadjuvant chemotherapy than luminal-like tumors. However, the specific genetic/epigenetic drivers of chemotherapy response among basal tumors are incompletely understood. Recently, gene expression subtypes were also found to correlate with response to immune checkpoint inhibition, including robust responses among a small but unique subset of patients with a neuronal tumor subtype. It is likely that applying similar genomic approaches to additional cohorts may provide further insights into the genomic drivers of therapeutic response and resistance to neoadjuvant therapies. Citation Format: Kent W. Mouw. Predictive biomarkers of response to neoadjuvant therapy [abstract]. In: Proceedings of the AACR Special Conference on Bladder Cancer: Transforming the Field; 2019 May 18-21; Denver, CO. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(15_Suppl):Abstract nr IA24.

Characterization of urinary extracellular vesicle proteins in muscle-invasive bladder cancer.

The mechanisms of bladder cancer progression are unknown, and new treatments and biomarkers are needed. Patient urinary extracellular vesicles (EVs) derive in part from bladder cancer cells and contain a specific protein cargo which may provide information about the disease. We conducted a proteomics study comparing EVs from the muscle-invasive bladder cancer (MIBC) cell line TCCSUP to EVs from normal urothelial line SVHUC. GO term analysis showed that TCCSUP EVs are enriched in proteins associated with the cell membrane, extracellular matrix, and inflammation and angiogenesis signaling pathways. Proteins characteristic of cancer EVs were further screened at the mRNA level in bladder cancer cell lines. In Western blots, three of six proteins examined showed greater than fifteenfold enrichment in patient urinary EVs compared to healthy volunteers (n = 6). Finally, we performed immunohistochemical staining of bladder tissue microarrays for three proteins of interest. One of them, transaldolase (TALDO1), is a nearly ubiquitous enzyme and normally thought to reside in the cytoplasm. To our surprise, nuclei were stained for transaldolase in 94% of MIBC tissue samples (n = 51). While cytoplasmic transaldolase was found in 89–90% of both normal urothelium (n = 79) and non-muscle-invasive samples (n = 71), the rate falls to 39% in MIBC samples (P < 0.001), and negative cytoplasmic staining was correlated with worse cancer-specific survival in MIBC patients (P = 0.008). The differential EV proteomics strategy reported here successfully identified a number of proteins associated with bladder cancer and points the way to future investigation.

Abstract PR18: Somatic ERCC2 mutations, nucleotide excision repair (NER) function, and cisplatin response in muscle-invasive bladder cancer (MIBC)

Abstract ERCC2 is a core member of the nucleotide excision repair (NER) pathway, a highly conserved and remarkably versatile DNA repair pathway responsible for repairing intrastrand DNA adducts created by genotoxic agents such as UV irradiation and platinum-based chemotherapies. Recent large-scale genomic efforts have shown that somatic ERCC2 missense mutations are present in approximately 20% of all primary muscle-invasive bladder cancers (MIBC). We previously showed that ERCC2 mutations are associated with treatment response and overall survival in MIBC patients treated with cisplatin-based chemotherapy. Initial functional studies on a subset of the observed ERCC2 mutations suggest that the mutations confer loss of normal cellular NER capacity. However, sequencing of additional MIBC cohorts has revealed that mutations occur across the ERCC2 gene, and the functional effects of the majority of these mutations remain unknown. In order to understand the functional landscape of ERCC2 mutations in MIBC, we have developed a high-throughput fluorescence-based assay to test the functional consequences of mutations in ERCC2 and other NER genes on cellular NER capacity. We apply this approach to all observed ERCC2 mutations across three published MIBC cohorts and find that the majority of ERCC2 mutations result in complete or near-complete loss of cellular NER. In addition, by correlating our functional results with available clinical data, we find interesting examples of cases in which ERCC2 status and cisplatin response are decoupled, highlighting the importance of using functional data to complement genomic and clinical endpoints in the search for reliable predictive biomarkers. This abstract is also being presented as Poster A29. Citation Format: Kent Mouw, Jean-Bernard Lazaro, Alexis Damish, Elizaveta Reznichenko, Zoe Frazier, David Liu, Jaegil Kim, Paz Polak, Levi Garraway, Gad Getz, Jonathan Rosenberg, Eliezer Van Allen, Alan D'Andrea. Somatic ERCC2 mutations, nucleotide excision repair (NER) function, and cisplatin response in muscle-invasive bladder cancer (MIBC) [abstract]. In: Proceedings of the AACR Special Conference on DNA Repair: Tumor Development and Therapeutic Response; 2016 Nov 2-5; Montreal, QC, Canada. Philadelphia (PA): AACR; Mol Cancer Res 2017;15(4_Suppl):Abstract nr PR18.

Overexpression of long non-coding RNA TUG1 predicts poor prognosis and promotes cancer cell proliferation and migration in high-grade muscle-invasive bladder cancer.

Long non-coding RNA TUG1 is involved in the development and progression of a variety of tumors. Little is known about TUG1 function in high-grade muscle-invasive bladder cancer (MIBC). The aims of our study were to determine expression levels of long non-coding RNA TUG1 in tumor tissue, to evaluate its relationship with clinico-pathological features of high-grade MIBC, and to describe its function in MIBC cells in vitro. TUG1 expression levels were determined in paired tumor and adjacent non-tumor bladder tissues of 47 patients with high-grade MIBC using real-time PCR. Cell line T-24 and siRNA silencing were used to study the TUG1 function in vitro. We observed significantly increased levels of TUG1 in tumor tissue in comparison to adjacent non-tumor bladder tissue (P<0.0001). TUG1 levels were significantly increased in metastatic tumors (P=0.0147) and were associated with shorter overall survival of MIBC patients (P=0.0241). TUG1 silencing in vitro led to 34% decrease in cancer cell proliferation (P=0.0004) and 23% reduction in migration capacity of cancer cells (P<0.0001). We did not observe any significant effects of TUG1 silencing on cell cycle distribution and number of apoptotic cells. Our study confirmed overexpression of TUG1 in MIBC tumor tissue and described its association with worse overall survival in high-grade MIBC patients. Together with in vitro observations, these data suggest an oncogenic role of TUG1 and its potential usage as biomarker or therapeutic target in MIBC.

Abstract 816: Genetic variants in the dopaminergic system and bladder cancer clinical outcomes

Abstract Bladder cancer is estimated to be the fourth most common cancer diagnosed and the eighth most common cancer related death in men in 2015. A hallmark of bladder cancer outcomes is the high rates of recurrence and progression in patients diagnosed with non-muscle invasive bladder cancer (NMIBC) and poor survival rates in those that have progressed to or are diagnosed with muscle invasive bladder cancer (MIBC). Because of the poor outcomes, research for predictors of clinical outcomes is necessary. Recent studies show that current depressive symptoms at time of bladder cancer diagnosis are associated with bladder cancer mortality. We are interested in investigating the genetic mechanism underlying this relationship. The dopaminergic pathway is of interest because of its associations with depression and smoking addiction (with smoking being the main risk factor for bladder cancer). Therefore we aimed to do a candidate pathway analysis examining 256 single nucleotide polymorphisms (SNPs) in 15 genes from the dopaminergic system. We analyzed the association of genetic variants with recurrence and progression in NMIBC patients (N = 497) and the association with survival in MIBC patients (N = 399). Overall, there were four SNPs with a p-value of less than 0.01associated with recurrence in NMIBC. The most significant variant being rs2797853 located in DBH under the recessive model (HR = 1.77, 95% CI: 1.28- 2.46. P = 0.0012). Eight polymorphisms had a statistically significant association with progression in NMIBC patients, with COMT: rs5993891 being the most significant variant showing an increased odds of bladder cancer progression (HR = 2.54, 95% CI: 1.50- 4.28, P = 0.0013). Interestingly, the variant rs174675 located in COMT (an enzyme responsible for degrading dopamine) was associated with an increased risk of recurrence (HR = 1.23, 95% CI: 1.01-1.49, P = 0.046) and progression (HR = 1.55, 95% CI: 1.13-2.12, P = 0.007). Finally, five SNPs were statistically significant for bladder cancer survival in MIBC patients. BDNF: rs2203877 was associated with a 71% increased risk of death (HR = 1.71, 95% CI: 1.25-2.34, P = 0.001). There was a significant difference in median survival time (MST). Those with the homozygous dominant or heterozygous genotype had a MST of 66.3 months and those with the homozygous recessive genotype had a MST of 31.8 months (Plog-rank = 0.02). The variant rs2239395 in COMT was associated with an increased risk of progression (HR = 2.54, 95% CI: 1.49-4.34, P = 0.002) and death (HR = 1.63, 95% CI: 1.05-2.53, P = 0.041). In addition to validation studies that are underway, the next steps include analyzing the association between these factors and BMI (a factor associated with depression, the dopamine system, and bladder cancer). The results of this study have the potential to contribute to identifying bladder cancer patients who may have poor clinical outcomes as well as the development of interventions that may improve outcomes. Citation Format: Jeanne A. Pierzynski, Yuanquing Ye, Alma Rodriguez, Michelle A.T. Hildebrandt, Ashish M. Kamat, Colin P.N. Dinney, Xifeng Wu. Genetic variants in the dopaminergic system and bladder cancer clinical outcomes. [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 816.