Bacillus Calmette-Guérin Treatment Research Articles

CyTOF analysis identifies unusual immune cells in urine of BCG-treated bladder cancer patients.

High grade non-muscle-invasive bladder tumours are treated with transurethral resection followed by recurrent intravesical instillations of Bacillus Calmette Guérin (BCG). Although most bladder cancer patients respond well to BCG, there is no clinical parameter predictive of treatment response, and when treatment fails, the prognosis is very poor. Further, a high percentage of NMIBC patients treated with BCG suffer unwanted effects that force them to stop treatment. Thus, early identification of patients in which BCG treatment will fail is really important. Here, to identify early stage non-invasive biomarkers of non-responder patients and patients at risk of abandoning the treatment, we longitudinally analysed the phenotype of cells released into the urine of bladder cancer patients 3-7 days after BCG instillations. Mass cytometry (CyTOF) analyses revealed a large proportion of granulocytes and monocytes, mostly expressing activation markers. A novel population of CD15+CD66b+CD14+CD16+ cells was highly abundant in several samples; expression of these markers was confirmed using flow cytometry and qPCR. A stronger inflammatory response was associated with increased cell numbers in the urine; this was not due to hematuria because the cell proportions were distinct from those in the blood. This pilot study represents the first CyTOF analysis of cells recruited to urine during BCG treatment, allowing identification of informative markers associated with treatment response for sub-selection of markers to confirm using conventional techniques. Further studies should jointly evaluate cells and soluble factors in urine in larger cohorts of patients to characterise the arms of the immune response activated in responders and to identify patients at risk of complications from BCG treatment.

FGFBP1 as a potential biomarker predicting bacillus Calmette–Guérin response in bladder cancer

Accurate prediction of Bacillus Calmette–Guérin (BCG) response is essential to identify bladder cancer (BCa) patients most likely to respond sustainably, but no molecular marker predicting BCG response is available in clinical routine. Therefore, we first identified that fibroblast growth factor binding protein 1 (FGFBP1) was upregulated in failures of BCG therapy, and the increased FGFBP1 had a poor outcome for BCa patients in the E-MTAB-4321 and GSE19423 datasets. These different expression genes associated with FGFBP1 expression are mainly involved in neutrophil activation, neutrophil-mediated immunity, and tumor necrosis factor-mediated signal pathways in biological processes. A significant positive correlation was observed between FGFBP1 expression and regulatory T-cell (Treg) infiltration by the Spearman correlation test in the BCG cohort (r = 0.177) and The Cancer Genome Atlas (TCGA) cohort (r = 0.176), suggesting that FGFBP1 may influence the response of BCa patients to BCG immunotherapy through immune escape. Though FGFBP1 expression was positively correlated with the expressions of PD-L1, CTLA4, and PDCD1 in TCGA cohort, a strong association between FGFBP1 and PD-L1 expression was only detected in the BCG cohort (r = 0.750). Furthermore, elevated FGFBP1 was observed in BCa cell lines and tissues in comparison to corresponding normal controls by RT-qPCR, Western blotting, and immunohistochemical staining. Increased FGFBP1 was further detected in the failures than in the responders by immunohistochemical staining. Notably, FGFBP1 is positively associated with PD-L1 expression in BCa patients with BCG treatment. To sum up, FGFBP1 in BCa tissue could be identified as a promising biomarker for the accurate prediction of BCG response in BCa.

The Role of Fluorescence In Situ Hybridization in the Surveillance of Non-Muscle Invasive Bladder Cancer: An Updated Systematic Review and Meta-Analysis.

Background: Fluorescence in situ hybridization (FISH) has become a popular biomarker for subsequent monitoring the recurrence of non-muscle invasive bladder cancer (NMIBC), several studies have investigated the ability of FISH to detect recurrence in the surveillance of NMIBC. However, the results were inconsistent. Methods: We conducted a systematic literature search extensively on authenticated databases including PubMed/Medline, Embase, Web of Science, Ovid, and Cochrane Library. Meta-analysis was performed to find out the sensitivity and specificity of FISH in predicting recurrence of NMIBC. Results: 15 studies were ultimately included in this meta-analysis, a total of 2941 FISH evaluations from 2385 NMIBC patients were available. The pooled sensitivity of FISH was 68% (95% CI: 0.58–0.76), and the pooled specificity was 64% (95% CI: 0.53–0.74). Subgroup analyses were performed in 7 studies without Bacillus Calmette–Guerin (BCG) treatment, the pooled sensitivity was 82% (95% CI: 0.68–0.90), and the pooled specificity was 63% (95% CI: 0.37–0.82). And in 9 studies using “UroVysion standard” to define positive FISH results showed a pooled sensitivity of 60% (95% CI: 0.50–0.70) and specificity of 70% (95% CI: 0.61–0.78). Conclusions: The findings of this study indicate that FISH has a satisfactory sensitivity (68%) and specificity (64%) and could be a potential biomarker in the surveillance of NMIBC. Moreover, BCG treatment and different FISH methods may have an impact on the sensitivity and specificity, these factors should be taken into account when making clinical strategy.

Castration immunoregulates toll-like receptor-4 in male bladder cancer.

Among diverse Pattern Recognition Receptors (PRRs), Toll-like receptor-4 (TLR-4) is a key urothelial trigger for innate immune response impacting urothelial bladder carcinoma (BC). Androgen activation promotes immunotolerance, playing an immunoregulatory role by unknown mechanisms. We explored the castration impact on urothelial TLR-4 modulation in carcinogenesis and immunotherapeutic scenario. Intact (SHAM) versus castrated male Fisher-344 rats were evaluated in 2 scenarios: (A) Carcinogenesis: After randomization to SHAM (n = 5) and Castration (n = 5), carcinogenesis was induced by four intravesical doses of 1.5mg/kg n-methyl-n-nitrosourea (MNU) every 15days. (B) Treatment: After ultrasonographic confirmed MNU-induced papillary BC on week 8, rats were randomized to SHAM (n = 5) and Castration (n = 5) and offered 6 weekly intravesical treatment of 106CFU of bacillus Calmette Guerin (BCG) in 0.2ml saline. After 15weeks the urinary bladders underwent histopathology. Urothelial cell proliferation was measured by Ki-67 immunohistochemistry (IHC), and TLR-4 expression was quantified by IHC and WB. Castration induced higher TLR-4 urothelial expression (p = 0.007) and anticarcinogenic effect with fewer urothelial tumors (60 vs. 80%) and lower urothelial cell proliferation compared to intact animals (p = 0.008). In the intravesical BCG treatment setting, castration has potentialized the BCG activation of TLR-4 (p = 0.007) with no residual in situ carcinoma compared to intact animals, suggesting the potential to amplify the BCG immune response. To our knowledge, this is the first description of TLR-4 urothelial expression hormonal modulation. The described castration-mediated immunomodulation will help to improve the knowledge of urothelial cancer gender diversities and PRRs modulations with treatment implications.

FACTORS PREDICTING THE OUTCOME OF BCG (BACILLUS CALMETTE-GUERIN) THERAPY

Background: Bacillus Calmette-Guerin is currently con­sidered to be the gold standard in conservative treatment for intermediate and high risk non-muscle invasive blad­der cancer (NMIBC). However, up to 40% of the patients, treated with adjuvant BCG therapy, experience failure. As the new methods of treatment after BCG failure ap­pear, there is a need to assess efficacy of this therapy in order to improve appropriate treatment during or before BCG therapy. Aim: To review the latest studies on factors, predicting BCG efficacy, and provide summarized information re­garding treatment outcomes in current practice. Methodology: The search was conducted in MEDLINE (Pubmed) and ScienceDirect databases using predeter­mined keywords: Bacillus Calmette-Guerin (BCG), fai­lure, intravesical therapy, outcome, response, predicting factors, immunological markers, efficacy. 10 articles pu­blished since 2017 were included in this literature review. Results: Prognostic factors such as the use of antibiotics prior to the therapy, body mass index (BMI), lympho­vascular invasion (LVI), endothelin-1 (ET-1), tumor substaging, CD4/CD8, GATA3/T-bet, CD163/CD68, neu­trophil-to-lymphocyte (NLR) ratios are promising factors for estimating the possible response of BCG therapy be­fore undergoing treatment. Blood eosinophil count and percentage, FISH test following 3 months after first BCG instillation and a Cytokine Panel for Response to Intrave­sical Therapy (CyPRIT) nomogram showed huge poten­tial for predicting BCG outcome during BCG treatment. Conclusion: There are quite many possible clinicopatho­logic, immunological, immunohistochemical and anam­nestical factors that could predict the success of BCG therapy. However, in order to include prognostic fac­tors in treatment guidelines, further studies should be performed to determine which of these factors have the greatest clinical value.

Lymphocyte-to-Monocyte Ratio (LMR) During Induction Is a Better Predictor Than Preoperative LMR in Patients Receiving Intravesical Bacillus Calmette -Guerin for Non-Muscle-Invasive Bladder Cancer.

The prognostic value of the lymphocyte-to-monocyte ratio during induction (ILMR) remains unclear in non-muscle-invasive bladder cancer (NMIBC) patients receiving Bacillus Calmette-Guérin (BCG). We aimed to determine and compare the prognostic value of the ILMR, preoperative lymphocyte-to-monocyte ratio (PLMR) and their dynamic changes (PILMR). This study collected the data from NMIBC patients receiving BCG treatment in our institution. The prognostic value of the PLMR, ILMR and PILMR was analyzed by the Kaplan−Meier method and Cox proportional hazard regression models. The concordance index and receiver operating characteristic curve analysis were employed to compare the prognostic value of these three factors. Our study enrolled 197 patients. These patients included 170 male patients, and the mean age was 64.17 years. During the follow-up time, 85 patients experienced recurrence, and 55 patients experienced progression. According to the results of COX multivariable analysis, PLMR (P=0.011) and ILMR (P<0.001) could independently predict the recurrence of NMIBC patients receiving BCG. Meanwhile, ILMR (P=0.001) and PILMR (P=0.036) were also the independent prognostic factors of progression. Compared with PLMR and PILMR, ILMR was associated with better accuracy for NMIBC patients receiving BCG. This study first found that the ILMR could independently predict the prognosis of NMIBC patients receiving BCG. Furthermore, we also identified that ILMR was associated with higher prognostic value than PLMR and PILMR, which might help to select an optimal treatment schedule for patients with NMIBC.

Abstract 1282: Elevated T cell exhaustion and immune cell infiltration is associated with BCG failure in patients with non-muscle invasive bladder cancer

Abstract Introduction: Gold standard treatment in patients with high-risk non-muscle invasive bladder cancer (NMIBC) includes Bacillus Calmette-Guérin (BCG), which activates the immune cells to kill remaining cancerous cells after surgical removal of the tumors. However, 40% of patients do not have a clinical benefit of BCG. The presence of immune cells, their functional state, as well as genomic alterations in the tumor and the normal appearing urothelial tissue, defined as field effect, may have significant impact on therapeutic outcome. Materials and methods: To investigate this, we analyzed samples from 156 patients diagnosed with NMIBC, including 237 tumors, 569 urine samples, and 305 biopsies of adjacent normal appearing urothelium. Patients had a median follow-up time of 8 years. 70 patients (45%) progressed to muscle-invasive bladder cancer or experienced early high grade recurrence within two years after ended BCG treatment, defined as BCG failure. Urinary levels of 92 immune-oncology related proteins were measured in pre- and post-treatment samples using the Olink proteomics platform. Total RNA- and whole exome sequencing (WES) data was generated from tumors before and after BCG. Clonal patient-specific mutations were selected for deep-targeted sequencing of the adjacent normal appearing biopsies. The level of field effect was defined as the mean of the variant allele frequency for mutations observed in normal biopsies. Results: We found that treatment with BCG activated the immune system regardless of clinical outcome. However, patients differed in urinary protein profile after BCG depending on their clinical response. Transcriptomic analysis of tumors showed that UROMOL2021 subtypes were significantly associated with outcome and clinical response (p=0.018). Paired tumors showed a shift in subtype to an immune infiltrated subtype (class 2b) after treatment in 36% of cases. Patients with BCG failure showed signs of immune exhaustion after treatment indicated by higher expression of the T cell exhaustion marker genes CTLA4 (p=0.0067), LAG3 (p=0.00012), TIM-3 (p=0.022), KLRG1 (p=0.028), and PD-1 (p=0.047), and higher immune cell infiltration (p=0.011). Genomic features such as mutational signatures and mutational load were not predictive of clinical response. Interestingly, BCG-responsive patients had a high level of field effect before BCG (p=0.0026), suggesting that field effect could lead to increased BCG efficacy. Conclusion: BCG treatment was associated with immune system activation reflected in both urine and tissue samples. In patients with BCG failure, we observed an immune exhausted phenotype after treatment, and we observed a high level of field effect before treatment in responsive patients. Collectively, this could indicate that increased immunogenicity and prolonged immune activation are key factors in BCG treatment responsiveness. Citation Format: Trine Strandgaard, Iver Nordentoft, Emil Christensen, Sia Lindskrog, Philippe Lamy, Karin Birkenkamp-Demtröder, Torben Steiniche, Jørgen Bjerggaard Jensen, Lars Dyrskjøt. Elevated T cell exhaustion and immune cell infiltration is associated with BCG failure in patients with non-muscle invasive bladder cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1282.

Abstract 5831: Urinary exosome analysis as a marker of treatment response in bladder cancer patients

Abstract Introduction: Intravesical Bacillus Calmette-Guerin (BCG) is an effective immunotherapy for non-muscle invasive bladder cancer (NMIBC). However, BCG treatment failure will lead to recurrence and tumor progression. In this study, urinary exosomes content (miRNA profile) was evaluated as a possible marker of BCG treatment response in NMIBC patients. Methods: Urine samples from patients with bladder cancer were collected at the time of surgery and during patient follow up to 1 year. Urine from healthy volunteers were also included. Clinical and pathological information such as tumor grade (High Grade (HG), Low Grade (LG)), depth of invasion (Ta, T1, CIS), and response to BCG treatment was also obtained. Exosome Isolation and total RNA extraction including microRNAs from cell-free urine after centrifugation were obtained. Library preparation for miRNA expression was done (QIAseq® miRNA Library) for Next-Generation Sequencing (NGS) analysis in a NextSeq 2000 single read platform, 75 bp with 15-20 million reads per sample. Reads were then queried against miRDeep2 software for identification. Only miRNAs having at least 20 counts considering all samples were included. After normalization, significantly and deferentially expressed microRNAs (&amp;gt;2-Fold) were selected for analysis. Bioinformatic analysis including sequence alignment was performed under the STAR-based approach. Identified microRNAs were then used to classify/predict the response to treatment and its relationship with other clinicopathologic variables. Results: A total of 56 urine samples from 13 patients were available/used for analysis including 10 High Grade Ta and 3 High Grade T1 patients. Urine from normal healthy donors (N=3) was also included. Clinicopathological features were patients with HGTa=10, HGT1=3 and 3 control samples. Regarding treatment response 9 patients were BCG responders and 4 BCG unresponsive. When compared to BCG unresponsive patients, BCG responders showed 45 differentially expressed miRNAs. Statistically significant differentially expressed miRNAs (Fold-change &amp;gt;2, p value &amp;lt;0.05) were 12 miRNAs, upregulated were miR132-3p (p=0.042); miR-187-3p (p=0.021); miR-409-3p (p=0.043) and miR1301-3p (p=0.048). Downregulated miRNAs were miR-let7-5p (p=0.007), miR-3605-3p (p=0.047), miR-140-5p (p=0.031), miR-500a-5p (p=0.051), miR-629-5p (p=0.039), miR-454-3p (p=0.05), miR-2110 (p=0.049) and miR-30c-5p (p=0.03). Interestingly, miRPathDABv2.0 analysis predicted those microRNAs be related to cancer, including bladder cancer. They showed targeting important pathways as PI3K.Akt, Wnt/B catenin and P53 signaling related with disease progression and treatment response. Conclusion: Our study supports the value of urinary exosomal microRNAs as non-invasive biomarkers to predict BCG treatment response in nonmuscle-invasive bladder cancer. Citation Format: Vladimir Valera, Beatriz Walter, Maria J. Merino. Urinary exosome analysis as a marker of treatment response in bladder cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5831.

Abstract 2058: Effect of route of Bacillus-Calmette Guerin administration on systemic immune responses in a murine model of non-muscle invasive bladder cancer

Abstract Background: Bladder cancer is the 5th most common cancer worldwide, with the most common type being non-muscle invasive bladder cancer (NMIBC). Treatment for high grade NMIBC includes resection of the bladder tumor followed by repeated intravesical administration of Bacillus-Calmette Guérin (BCG), the live-attenuated form of Mycobacterium bovis. Unfortunately, 60-70% of patients will experience disease recurrence. Our current understanding is that BCG acts within the local bladder tumor microenvironment and draining lymph nodes, leading to the activation of a systemic immune response. Using a murine model of NMIBC, here we investigated whether different routes of BCG administration alter the systemic immune response to BCG and affect tumor size. Methods: To establish syngeneic orthotopic bladder tumors in a mouse model, female C57Bl/6 mice were catheterized and 2.5 x 105 mouse bladder cancer cells (MB49) were instilled into the bladder. Beginning on day 7, mice received either intravesical, intravenous (IV) or intraperitoneal (IP) BCG (8 mg/ml) or saline once weekly for three weeks and bladders were imaged through ultrasound. At endpoint (day 23), tumors, tumor draining lymph nodes (tdLNs), spleen, and bone marrow were harvested and stored. Immune composition of the various lymphoid organs was evaluated using polychromatic flow cytometry. Ex vivo responsiveness to lipopolysaccharide (LPS) was assessed in bone marrow cells by measuring cytokine levels in culture supernatants. Results: Mice treated with IV BCG had significantly smaller tumor volumes when compared to intravesical BCG treatment. Ex vivo LPS stimulation of bone marrow from BCG-treated mice, regardless of administration route, led to increased release of pro-inflammatory cytokines compared with saline controls. IP and IV BCG treatment skewed immune microenvironments of secondary lymphoid organs towards a cytotoxic phenotype compared with intravesical BCG treatment. Within the lymphoid compartment of tdLNs, the proportion of CD8+ T cells was significantly increased while the proportion of CD4+ T cells was significantly decreased in mice treated with IV and IP BCG compared with intravesical BCG. Conversely, the proportion of CD11b+ dendritic cells (DCs) was significantly increased in tdLNs from mice receiving intravesical BCG compared with BCG administered IV or IP. Within the spleen, the proportion of CD8+CD11b- DCs were significantly increased in the DC compartment following IP BCG treatment compared to other treatment routes. Conclusion: This provides evidence that route of BCG administration affects the immune composition of secondary lymphoid organs that may be associated with anti-tumor responses. A better understanding of the mechanism of BCG immunotherapy will lead to novel approaches to optimize anti-tumor immune responses and, consequently, improved patient outcomes. Citation Format: Arielle Grossman, Aline Atallah, Tiziana Cotechini, Jean-Francois Pare, Robert Siemens, Charles H. Graham. Effect of route of Bacillus-Calmette Guerin administration on systemic immune responses in a murine model of non-muscle invasive bladder cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2058.

Abstract 3540: Effect of route of Bacillus Calmette Guerin administration on the immune microenvironment and growth of bladder tumors

Abstract Background: Bladder cancer is the fifth most common cancer in North America. The standard of care for high-risk non-muscle invasive bladder cancer (NMIBC) involves intravesical immunotherapy with Bacillus Calmette Guérin (BCG). However, it is possible that the efficacy of this modality of BCG administration is suboptimal, as most patients do not respond fully to this immunotherapy. Furthermore, our current understanding of the immunotherapeutic effect of BCG is incomplete. Using a mouse model of NMIBC, we compared the tumor immune microenvironment (TiME) following intravesical versus intravenous administration of BCG, as well as changes in tumor volume and mice survival. Methods: Female C57Bl/6 mice (6-8 weeks old) were catheterized and instilled with 2.5x105 MB49 bladder cancer cells after poly-L-lysine treatment. Beginning on day 7, mice were treated with intravenous (IV) or intravesical BCG (8 mg/ml) or saline once weekly for three weeks. Ultrasound was performed weekly to monitor tumor growth. Similarly, a cohort of non-tumor bearing mice was treated with poly-L-lysine on day one followed by three weekly intravesical instillations of BCG. In both cohorts, mice were sacrificed on day 23 and bladders were harvested, enzymatically dispersed to generate single-cell suspensions for analysis by flow cytometry, or snap frozen for transcriptomic analysis using NanoString nCounter Platform. Results: Mice receiving IV BCG had better survival and their bladder tumors were significantly smaller compared with mice receiving intravesical BCG. This reduction in tumor size was associated with a significantly increased proportion of CD8+ T cells and a significantly reduced proportion of inflammatory monocytes in bladder tumors from mice treated with IV BCG compared with intravesical BCG. Whole tumor transcriptome analysis revealed alterations in various signalling pathways associated with route of BCG administration. Our results demonstrate similar trends in the distribution of immune populations in the TiME following intravesical saline and IV BCG treatment, as well as following IV saline and intravesical BCG treatment. We also found that the TiME after intravesical BCG treatment has more immune cells which are predominantly immature myeloid cells. Moreover, our results indicate that intravesical BCG treatment leads to a significant population of immature myeloid cells in the bladders of non-tumor bearing mice. Conclusion/Significance: These results provide evidence that the route of BCG administration is an important determinant of the TiME composition and may influence anti-tumor responses. Understanding the link between the TiME and BCG therapy may facilitate the development of new approaches to improve outcomes and reduce recurrence rates in patients with NMIBC. Citation Format: Aline Atallah, Arielle Grossman, Jean-Francois Pare, Robert Siemens, Tiziana Cotechini, Charles H. Graham. Effect of route of Bacillus Calmette Guerin administration on the immune microenvironment and growth of bladder tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3540.

Assessment of changes in the international prostate symptom score for patients enrolled in a phase I trial of intravesical bacillus Calmette-Guérin combined with intravenous pembrolizumab for recurrent or persistent high-grade non-muscle-invasive bladder cancer after previous bacillus Calmette-Guérin treatment.

e16553 Background: We conducted the first phase I dose-escalation trial (NCT02324582) of intravesical Bacillus Calmette-Guérin (BCG) in combination with systemic pembrolizumab in patients with high-grade non-muscle-invasive bladder cancer who had persistent or recurrent disease after prior intravesical therapy with BCG. The International Prostate Symptom Score (IPSS) were administered to assess subjects’ urinary bother during the trial. Methods: Eighteen subjects were consented for the study, 13 were treated. The study population was comprised of 11 males, 2 females, with age range 42 - 92 years. The IPSS was assessed at pre-treatment, Week 10 (during combined BCG and pembrolizumab treatment), and at 3 and 6 months from treatment completion. IPSS outcomes were analyzed using a mixed-model repeated measures analysis with p-value &lt; 0.05 considered statically significant. Results: For each time point, there was no significant change in any of the component variables of the IPSS. There was no increase in reported bother symptoms over the course of treatment, given median IPSS scores of 9, 11, 7, and 8 (IQR 1.75) for each time point, respectively. Furthermore, quality of life measurements demonstrated no significant change in subjective burden with 75% of subjects reporting that they were “mostly satisfied” with treatment outcomes, followed by 13% reporting “delighted” or “pleased” at 6 month follow-up. Conclusions: There is no significant change in quality of life or the perceived bother of symptoms indicated in the component variables of the IPSS for our study population before, during or after treatment.

Expression of RPL9 predicts the recurrence of non-muscle invasive bladder cancer with BCG therapy

Numerous biomarkers and risk tables can be used to predict recurrence or progression of patients with primary or recurrent non-muscle invasive bladder cancer (NMIBC) receiving Bacillus Calmette-Guerin (BCG). However, few are suitable for BCG-unresponsive disease (i.e., recurrence or progression after BCG treatment). Therefore, identification of a novel marker that allows accurate prediction of prognosis, particularly risk of recurrence, is critically important in clinical practice. In the current study, gene ontology and gene set enrichment analyses of microarray datasets (GSE13507, n = 47) revealed that differentially expressed genes in recurred NMIBC patients after BCG treatment were associated with virus and ribosomal pathways. Among the core-enrichment genes, the expression of RPL9, a putative tumor suppressor, was lower in recurred NMIBC patients after BCG therapy than in patients without recurrence (P = 0.033) from the E-MTAT-4321 European cohort (n = 84). Data from The Cancer Genome Atlas (n = 406) showed that bladder cancer patients with higher RPL9 expression had a longer overall survival probability than patients with lower RPL9 expression (P = 0.011). Moreover, we used the latest digital PCR platform to examine 59 NMIBC patients and identified downregulation of RPL9 in patients with recurrence after BCG therapy (P = 0.031). The Kaplan-Meier survival estimator showed that NMIBC patients with higher expression of RPL9 had longer recurrence-free survival (log-rank test, P = 0.015). Therefore, we conclude that RPL9 expression is a prospective predictor of recurrence after BCG therapy in NMIBC patients.

MP59-06 PD-L1 EXPRESSION AND BCG RESPONSE IN NON-MUSCLE INVASIVE BLADDER CANCER

You have accessJournal of UrologyCME1 May 2022MP59-06 PD-L1 EXPRESSION AND BCG RESPONSE IN NON-MUSCLE INVASIVE BLADDER CANCER Solomon Woldu, Thomas Gerald, Vitaly Margulis, Daniel Halstuch, Yaara Ber, Karin Lifshitz, David Margel, Yair Lotan, and Liwei Jia Solomon WolduSolomon Woldu More articles by this author , Thomas GeraldThomas Gerald More articles by this author , Vitaly MargulisVitaly Margulis More articles by this author , Daniel HalstuchDaniel Halstuch More articles by this author , Yaara BerYaara Ber More articles by this author , Karin LifshitzKarin Lifshitz More articles by this author , David MargelDavid Margel More articles by this author , Yair LotanYair Lotan More articles by this author , and Liwei JiaLiwei Jia More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002642.06AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Intravesical bacillus Calmette Guerin (BCG) is the standard of care adjuvant therapy for high risk non-muscle invasive bladder cancer (NMIBC), yet many patients experience recurrence or disease progression. The mechanism of BCG is thought to be related to stimulation of immune surveillance. Relatedly, systemic immune checkpoint inhibition is being utilized in advanced bladder cancer and approved for BCG unresponsive NMIBC. We sought to determine the association between PD-L1 expression and BCG treatment. METHODS: We identified 102 BCG-naïve patients with high grade (HG) NMIBC treated with BCG. All patients underwent initial transurethral resection (TUR). Dako 22c3 assay was used to determine PD-L1 expression. Patients were defined as PD-L1 positive if the combined positive score > 0. BCG unresponsiveness was defined by presence of HG disease at 6 months following adequate BCG (one induction and maintenance cycle or two induction cycles) for pT1 or 12 months for CIS or presence of pT1 at 3 months after induction. HG relapse was defined as HG disease after being followed for 6 months after BCG. RESULTS: The median follow-up was 57 months. Median number of BCG maintenance cycles was 1, and 17 (16.7%) patients underwent immediate reinduction BCG. PD-L1 expression was observed in 5.9% of pTa, 30.0% of pT1, and 3.6% of CIS. BCG unresponsiveness and HG relapse were observed in 32 (35.6%) and 29 (34.5%), respectively. On univariate analysis, PD-L1 expression was inversely associated with BCG unresponsiveness (OR=0.11; 95% CI 0.01-0.90) but not HG relapse (OR=0.30; 95% CI 0.06-1.44). The sensitivity, specificity, positive predictive value, and negative predictive value of PD-L1 expression for BCG responsiveness were 22%, 97%, 93%, and 41%, respectively. PD-L1 positive patients had a post-test probability of BCG responsiveness of 93%. On multivariate regression, pT1 (OR = 0.16; 95% CI 0.05-0.60), CIS (OR=0.25; 95% CI 0.07-0.86), and PD-L1 expression (OR=15.63; 95% CI 1.78-142.86) were associated with BCG responsiveness. CONCLUSIONS: PD-L1 expression in HG NMIBC was low, and those with PD-L1 expression at initial TUR were more likely to harbor invasive disease. Patients with PD-L1 expression were more likely to demonstrate BCG responsiveness. These findings suggest a role of PD-L1 in the immune surveillance mechanism of BCG at initial diagnosis and may assist in predicting response to BCG in patients with HG NMIBC. Further investigation is needed to determine if additional immune checkpoint markers have strong correlation with BCG response, particularly among patients without PD-L1 expression. Source of Funding: None © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e1003 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Solomon Woldu More articles by this author Thomas Gerald More articles by this author Vitaly Margulis More articles by this author Daniel Halstuch More articles by this author Yaara Ber More articles by this author Karin Lifshitz More articles by this author David Margel More articles by this author Yair Lotan More articles by this author Liwei Jia More articles by this author Expand All Advertisement PDF DownloadLoading ...

MP40-05 RISK FACTORS OF SUBSEQUENT UPPER URINARY TRACT CARCINOMA TREATED WITH INTRAVESICAL BACILLUS CALMETTE-GUERIN THERAPY FOR NON-MUSCLE-INVASIVE BLADDER CANCER: FROM MULTIPLE JAPANESE HOSPITALS

You have accessJournal of UrologyCME1 May 2022MP40-05 RISK FACTORS OF SUBSEQUENT UPPER URINARY TRACT CARCINOMA TREATED WITH INTRAVESICAL BACILLUS CALMETTE-GUERIN THERAPY FOR NON-MUSCLE-INVASIVE BLADDER CANCER: FROM MULTIPLE JAPANESE HOSPITALS Kazuyuki Numakura, Makito Miyake, Yuya Sekine, Yumina Muto, Mizuki Kobayashi, Rikiya Taoka, Takashi Kobayasi, Takahiro Kojima, Hiroshi Kitamura, Hiroyuki Nishiyama, Kiyohide Fujimoto, and Tomonori Habuchi Kazuyuki NumakuraKazuyuki Numakura More articles by this author , Makito MiyakeMakito Miyake More articles by this author , Yuya SekineYuya Sekine More articles by this author , Yumina MutoYumina Muto More articles by this author , Mizuki KobayashiMizuki Kobayashi More articles by this author , Rikiya TaokaRikiya Taoka More articles by this author , Takashi KobayasiTakashi Kobayasi More articles by this author , Takahiro KojimaTakahiro Kojima More articles by this author , Hiroshi KitamuraHiroshi Kitamura More articles by this author , Hiroyuki NishiyamaHiroyuki Nishiyama More articles by this author , Kiyohide FujimotoKiyohide Fujimoto More articles by this author , and Tomonori HabuchiTomonori Habuchi More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002600.05AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Patients with urothelial cancer experience frequent asynchronous and multi-locational recurrence. An upper urinary tract urothelial carcinoma (UTUC) is rare after intravesical bacillus Calmette-Guerin (BCG) therapy, and its incidence, clinical impacts, and risk factors have not been fully understood. Our study aimed to elucidate the clinical impact of UTUC after intravesical BCG therapy. METHODS: This retrospective cohort study used data collected between December 2017 and January 2000 from multiple Japanese hospitals (JUOG-UC 1901). Research involving human participants was approved by the Ethics board of all hospitals. A total of 3,226 patients diagnosed with non-muscle-invasive bladder cancer (NMIBC) and treated with intravesical BCG therapy were enrolled. We evaluated the impact of UTUC by comparing the intravesical recurrence-free (RFS), cancer-specific (CSS), and overall survival (OS) rates. The predictor of UTUC after BCG was assessed by multivariate logistic regression analysis. RESULTS: The median follow-up period from BCG treatment after transurethral resection of the bladder tumor (TURBT) was 48.0 months (IQR: 26.0–76.0). UTUC was detected in 175 (5.4%) patients during follow-up. Patients with UTUC had worse survival rates than those without UTUC: RFS (70.5 m vs 147.3 m, p <0.001), CSS (168.3 m vs 227.4 m, p <0.001), and OS (156.0 vs 189.6, p=0.019) (Figure 1). Multivariate analyses revealed that tumor multiplicity (OR, 1.681; 95% CI, 1.005–2.812; p=0.048), Connaught strain (OR, 2.211; 95% CI, 1.380–3.543; p=0.001), and intravesical recurrence (OR, 5.097; 95% CI, 3.225–8.056; p <0.001) were associated with UTUC after BCG therapy (Table 1). CONCLUSIONS: Patients with recurrent UTUC showed worse survival rates in terms of RFS, CSS, and OS than those without UTUC. Multiple primary bladder tumors, Connaught strain, and intravesical recurrence after BCG may be predictive factors for UTUC recurrence. Source of Funding: none © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e675 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Kazuyuki Numakura More articles by this author Makito Miyake More articles by this author Yuya Sekine More articles by this author Yumina Muto More articles by this author Mizuki Kobayashi More articles by this author Rikiya Taoka More articles by this author Takashi Kobayasi More articles by this author Takahiro Kojima More articles by this author Hiroshi Kitamura More articles by this author Hiroyuki Nishiyama More articles by this author Kiyohide Fujimoto More articles by this author Tomonori Habuchi More articles by this author Expand All Advertisement PDF DownloadLoading ...

MP59-02 APPLICATION OF A MULTIPLEX URINALYSIS TEST FOR THE PREDICTION OF INTRAVESICAL BCG TREATMENT RESPONSE: A PILOT STUDY

You have accessJournal of UrologyCME1 May 2022MP59-02 APPLICATION OF A MULTIPLEX URINALYSIS TEST FOR THE PREDICTION OF INTRAVESICAL BCG TREATMENT RESPONSE: A PILOT STUDY Kaoru Murakami, Ashish M. Kamat, Yunfeng Dai, Ian Pagano, Runpu Chen, Yijun Sun, Amit Gupta, Nari Kim, Steve Goodison, Charles J. Rosser, and Hideki Furuya Kaoru MurakamiKaoru Murakami More articles by this author , Ashish M. KamatAshish M. Kamat More articles by this author , Yunfeng DaiYunfeng Dai More articles by this author , Ian PaganoIan Pagano More articles by this author , Runpu ChenRunpu Chen More articles by this author , Yijun SunYijun Sun More articles by this author , Amit GuptaAmit Gupta More articles by this author , Nari KimNari Kim More articles by this author , Steve GoodisonSteve Goodison More articles by this author , Charles J. RosserCharles J. Rosser More articles by this author , and Hideki FuruyaHideki Furuya More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002642.02AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Intravesical Bacillus Calmette-Guerin (BCG), a live attenuated tuberculosis vaccine that acts as a non-specific immune system stimulant, is the most effective adjuvant treatment for patients with intermediate or high-risk non-muscle-invasive bladder cancer (NMIBC). However, to date, there are no reliable tests that are predictive of BCG treatment response. In this study, we evaluated the performance of OncuriaTM, a bladder cancer detection test, to predict response to intravesical BCG. METHODS: OncuriaTM data was evaluated in voided urine samples obtained from a prospectively collected cohort of 64 subjects with intermediate or high risk NMIBC prior to treatment with intravesical BCG. The OncuriaTM test, which measures 10 cancer-associated biomarkers was performed in an independent clinical laboratory. The ability of the test to identify those patients in whom BCG is ineffective against tumor recurrence was tested. Predictive models were derived using supervised learning and cross-validation analyses. Model performance was assessed using ROC curves. RESULTS: Pre-treatment urinary concentrations of MMP9, VEGFA, CA9, SDC1, PAI1, APOE, A1AT, ANG and MMP10 were increased in patients who developed disease recurrence. A combinatorial predictive model of treatment outcome achieved an AUROC 0.89 [95% CI: 0.80–0.99], outperforming any single biomarker, with a test sensitivity of 81.8% and a specificity of 84.9%. Hazard ratio analysis revealed that patients with higher urinary levels of ANG, CA9 and MMP10 had a significantly higher risk of disease recurrence. CONCLUSIONS: Monitoring the urinary levels of a cancer-associated biomarker panel enabled the discrimination of patients who did not respond to intravesical BCG therapy. With further study, the multiplex OncuriaTM test may be applicable for the clinical evaluation of bladder cancer patients considering intravesical BCG treatment. We are currently performing a validation study with a larger retrospective cohort of ∼120 subjects with intermediate or high risk NMIBC prior to treatment with intravesical BCG. The results will be presented in the AUA 2022. Source of Funding: NIH/NCI R01CA198887 (CJR) © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e1001 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Kaoru Murakami More articles by this author Ashish M. Kamat More articles by this author Yunfeng Dai More articles by this author Ian Pagano More articles by this author Runpu Chen More articles by this author Yijun Sun More articles by this author Amit Gupta More articles by this author Nari Kim More articles by this author Steve Goodison More articles by this author Charles J. Rosser More articles by this author Hideki Furuya More articles by this author Expand All Advertisement PDF DownloadLoading ...

PD12-11 CASTRATION IMPACT ON PATTERN RECOGNITION RECEPTORS (PRRS) BOOSTS TOLL-LIKE RECEPTOR-4 EXPRESSION IN AN IMMUNOCOMPETENT BLADDER CANCER MALE MODEL

You have accessJournal of UrologyCME1 May 2022PD12-11 CASTRATION IMPACT ON PATTERN RECOGNITION RECEPTORS (PRRS) BOOSTS TOLL-LIKE RECEPTOR-4 EXPRESSION IN AN IMMUNOCOMPETENT BLADDER CANCER MALE MODEL Leonardo Reis Leonardo ReisLeonardo Reis More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002538.11AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Among diverse Pattern Recognition Receptors (PRRs), the Toll-Like Receptor-4 (TLR-4) is the key urothelial trigger for innate immune response and plays a central role as Bacillus Calmette Guerin (BCG) main target. Also, its gene polymorphism and expression are associated to bladder cancer (BCa) prognosis. Androgen activation promotes immunotolerance, playing an immunoregulatory role, though by unknown mechanisms. METHODS: We described an efficient male immunocompetent BCa animal model. Intact (sham) versus castrated male Fisher-344 rats were evaluated in 2 scenarios: A) Carcinogenesis: After randomization to SHAM (n=5) and Castration (n=5), carcinogenesis was induced by four intravesical doses of 1.5 mg/kg n-methyl-n-nitrosourea (MNU) every 15 days. B) Treatment: After ultrasonographic confirmed MNU induced papillary BCa on week 8, rats were randomized to SHAM (n=5) and Castration (N=5) and offered 6 weekly intravesical treatment of 106 CFU of BCG in 0.2 ml saline. After 15 weeks of protocol, animals were euthanized and the urinary bladders submitted to histopathology, immunohistochemistry (IHC), and Western blotting (WB). Urothelial cell proliferation was measured by Ki-67 IHC and TLR-4 expression quantified by IHC and WB. RESULTS: Castration induced higher TLR-4 urothelial expression (p=.007) and anticarcinogenic effect with less urothelial tumors (60 vs. 80%) and lower urothelial cell proliferation compared to intact animals (p=.008). In the intravesical BCG treatment setting, castration has potentialized the BCG activation of TLR-4 (p=.007) with no residual in situ carcinoma compared to intact animals, suggesting the potential to amplify the immune initiation phase activated by BCG. CONCLUSIONS: To our knowledge, this is the first description of TLR-4 urothelial expression hormonal modulation. The described male immunocompetent animal model will help to improve the knowledge of urothelial cancer gender diversities and PRRs modulations with treatment implications. Source of Funding: none © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e199 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Leonardo Reis More articles by this author Expand All Advertisement PDF DownloadLoading ...

Bacillus Calmette-Guerin (BCG) induces superior anti-tumour responses by Vδ2+ T cells compared with the aminobisphosphonate drug zoledronic acid.

Vδ2+ T cells can recognize malignantly transformed cells as well as those infected with mycobacteria. This cross-reactivity supports the idea of using mycobacteria to manipulate Vδ2+ T cells in cancer immunotherapy. To date, therapeutic interventions using Vδ2+ T cells in cancer have involved expanding these cells in or ex vivo using zoledronic acid (ZA). Here, we show that the mycobacterium Bacillus Calmette–Guérin (BCG) also causes Vδ2+ T-cell expansion in vitro and that resulting Vδ2+ cell populations are cytotoxic toward tumour cell lines. We show that both ZA and BCG-expanded Vδ2+ cells effectively killed both Daudi and THP-1 cells. THP-1 cell killing by both ZA and BCG-expanded Vδ2+ cells was enhanced by treatment of targets cells with ZA. Although no difference in cytotoxic activity between ZA- and BCG-expanded Vδ2+ cells was observed, BCG-expanded cells degranulated more and produced a more diverse range of cytokines upon tumour cell recognition compared to ZA-expanded cells. ZA-expanded Vδ2+ cells were shown to upregulate exhaustion marker CD57 to a greater extent than BCG-expanded Vδ2+ cells. Furthermore, ZA expansion was associated with upregulation of inhibitory markers PD-1 and TIM3 in a dose-dependent manner whereas PD-1 expression was not increased following expansion using BCG. Intradermal BCG vaccination of rhesus macaques caused in vivo expansion of Vδ2+ cells. In combination with the aforementioned in vitro data, this finding suggests that BCG treatment could induce expansion of Vδ2+ T cells with enhanced anti-tumour potential compared to ZA treatment and that either ZA or BCG could be used intratumourally as a means to potentiate stronger anti-tumour Vδ2+ T-cell responses.

Response and Outcomes to Immune Checkpoint Inhibitors in Advanced Urothelial Cancer Based on Prior Intravesical Bacillus Calmette-Guerin.

Immune checkpoint inhibitors (ICI) improve overall survival (OS) in patients with locally advanced, unresectable, or metastatic urothelial carcinoma (aUC), but response rates can be modest. We compared outcomes between patients with and without prior intravesical Bacillus Calmette-Guerin (BCG), who received ICI for aUC, hypothesizing that prior intravesical BCG would be associated with worse outcomes. We performed a retrospective cohort study across 25 institutions in US and Europe. We compared observed response rate (ORR) using logistic regression; progression-free survival (PFS) and OS using Kaplan-Meier and Cox proportional hazards. Analyses were stratified by treatment line (first line/salvage) and included multivariable models adjusting for known prognostic factors. A total of 1026 patients with aUC were identified; 614, 617, and 638 were included in ORR, OS, PFS analyses, respectively. Overall, 150 pts had history of prior intravesical BCG treatment. ORR to ICI was similar between those with and without prior intravesical BCG exposure in both first line and salvage settings (adjusted odds radios 0.55 [P= .08] and 1.65 [P= .12]). OS (adjusted hazard ratios 1.05 [P= .79] and 1.13 [P= .49]) and PFS (adjusted hazard ratios 1.12 [P= .55] and 0.87 [P= .39]) were similar between those with and without intravesical BCG exposure in first line and salvage settings. Prior intravesical BCG was not associated with differences in response and survival in patients with aUC treated with ICI. Limitations include retrospective nature, lack of randomization, presence of selection and confounding biases. This study provides important preliminary data that prior intravesical BCG exposure may not impact ICI efficacy in aUC.

Prognostic and Predictive Value of Fibroblast Growth Factor Receptor Alterations in High-grade Non–muscle-invasive Bladder Cancer Treated with and Without Bacillus Calmette-Guérin Immunotherapy

BackgroundLimited data are available on the prognostic and predictive value of fibroblast growth factor receptor alterations (FGFRa) relative to clinical outcomes in patients with non–muscle-invasive bladder cancer (NMIBC). ObjectiveTo determine whether FGFRa may be clinically useful in stratifying for treatment response in a real-world cohort of patients with pT1 NMIBC treated and untreated with bacillus Calmette-Guérin (BCG) instillation therapy. Design, setting, and participantsA pooled dataset of matched clinical and genomic data (1992–2015) for pT1 NMIBC patients was assessed by the Bladder Cancer Research Initiative for Drug Targets in Germany consortium. Outcome measurements and statistical analysisKey efficacy outcomes were recurrence-free survival (RFS), progression-free survival (PFS), and disease-specific survival (DSS), which were estimated by Kaplan-Meier analysis, with hazard ratios calculated using a multivariate Cox proportional-hazard model. Results and limitationsIn this retrospective study of 263 patients with high-grade NMIBC, at a median follow-up of 63 mo, 32% showed recurrence and 15% progressed to muscle-invasive bladder cancer. FGFRa were found in 43% of patients, including 39% mutations and 5.7% fusions. FGFRa were associated with lower rates of concomitant carcinoma in situ. Among patients with or without FGFRa, there was no significant difference in PFS, RFS, and DSS in those who were BCG treated or BCG naive, or in the overall population. Limitations include the retrospective design from a single-center setting. ConclusionsIn patients with high-risk NMIBC, FGFRa were frequently observed. Patients with FGFRa who often exhibit recurrence/relapse after BCG treatment have a high unmet need. Patient summaryOur retrospective study suggests that fibroblast growth factor receptor alterations (FGFRa) occur frequently in non–muscle-invasive bladder cancer (NMIBC). Outcomes were similar with or without FGFRa in patients with NMIBC, both overall and for standard bacillus Calmette-Guérin (BCG) treatment.

Radiomics of Contrast-Enhanced Computed Tomography: A Potential Biomarker for Pretreatment Prediction of the Response to Bacillus Calmette-Guerin Immunotherapy in Non-Muscle-Invasive Bladder Cancer.

Background: Bacillus Calmette-Guerin (BCG) instillation is recommended postoperatively after transurethral resection of bladder cancer (TURBT) in patients with high-risk non-muscle-invasive bladder cancer (NMIBC). An accurate prediction model for the BCG response can help identify patients with NMIBC who may benefit from alternative therapy. Objective: To investigate the value of computed tomography (CT) radiomics features in predicting the response to BCG instillation among patients with primary high-risk NMIBC. Methods: Patients with pathologically confirmed high-risk NMIBC were retrospectively reviewed. Patients who underwent contrast-enhanced CT examination within one to 2 weeks before TURBT and received ≥5 BCG instillation treatments in two independent hospitals were enrolled. Patients with a routine follow-up of at least 1 year at the outpatient department were included in the final cohort. Radiomics features based on CT images were extracted from the tumor and its periphery in the training cohort, and a radiomics signature was built with recursive feature elimination. Selected features further underwent an unsupervised radiomics analysis using the newly introduced method, non-negative matrix factorization (NMF), to compute factor factorization decompositions of the radiomics matrix. Finally, a robust component, which was most associated with BCG failure in 1 year, was selected. The performance of the selected component was assessed and tested in an external validation cohort. Results: Overall, 128 patients (training cohort, n = 104; external validation cohort, n = 24) were included, including 12 BCG failures in the training cohort and 11 failures in the validation cohort each. NMF revealed five components, of which component 3 was selected for the best discrimination of BCG failure; it had an area under the curve (AUC) of .79, sensitivity of .79, and specificity of .65 in the training set. In the external validation cohort, it achieved an AUC of .68, sensitivity of .73, and specificity of .69. Survival analysis showed that patients with higher component scores had poor recurrence-free survival (RFS) in both cohorts (C-index: training cohort, .69; validation cohort, .68). Conclusion: The study suggested that radiomics components based on NMF might be a potential biomarker to predict BCG response and RFS after BCG treatment in patients with high-risk NMIBC.