Immunotherapy For Bladder Cancer Research Articles

Gut microbiota Parabacteroides distasonis enchances the efficacy of immunotherapy for bladder cancer by activating anti-tumor immune responses

ObjectiveBladder cancer(BCa) was a disease that seriously affects patients’ quality of life and prognosis. To address this issue, many researches suggested that the gut microbiota modulated tumor response to treatment; however, this had not been well-characterized in bladder cancer. In this study, our objective was to determine whether the diversity and composition of the gut microbiota or the density of specific bacterial genera influence the prognosis of patients with bladder cancer.MethodsWe collected fecal samples from a total of 50 bladder cancer patients and 22 matched non-cancer individuals for 16S rDNA sequencing to investigate the distribution of Parabacteroides in these two groups. Further we conducted follow-up with cancer patients to access the impact of different genera of microorganisms on patients survival. We conducted a Fecal Microbiota Transplantation (FMT) and mono-colonization experiment with Parabacteroides distasonis to explore its potential enhancement of the efficacy of anti-PD-1 immunotherapy in MB49 tumor-bearing mice. Immunohistochemistry, transcriptomics and molecular experiment analyses were employed to uncover the underlying mechanisms.ResultsThe 16S rDNA showed that abundance of the genus Parabacteroides was elevated in the non-cancer control group compared to bladder cancer group. The results of tumor growth curves showed that a combination therapy of P. distasonis and ICIs treatment significantly delayed tumor growth and increased the intratumoral densities ofboth CD4+T and CD8+T cells. The results of transcriptome analysis demonstrated that the pathways associated with antitumoral immune response were remarkably upregulated in the P. distasonis gavage group.ConclusionP. distasonis delivery combined with α-PD-1 mAb could be a new strategy to enhance the effect of anti-PD-1 immunotherapy. This effect might be achieved by activating immune and antitumor related pathways.

Chemotherapy, immunotherapy, or combination first-line treatment for metastatic urothelial carcinoma of the bladder: A large real-world experience

IntroductionFirst-line systemic therapy for metastatic urothelial carcinoma of the bladder (mUC) consists of platinum-based chemotherapy in most patients and PD1/L1 inhibitors in selected patients. Multiple combination chemoimmunotherapy trials failed to show a clear benefit over chemotherapy alone. We used real-world data to evaluate clinical and sociodemographic factors associated with receipt of first-line chemotherapy, immunotherapy, or combination chemoimmunotherapy treatment for metastatic bladder cancer and examined differences in overall survival (OS). Materials and MethodsWe used the National Cancer Database to identify patients with stage IV mUC diagnosed between 2014 and 2018, who were treated with first-line immunotherapy, chemotherapy, or combination treatment. We performed multivariable logistic regression modeling to determine factors associated with treatment receipt Adjusted Kaplan-Meier survival analysis and multivariable Cox proportional hazards regression were used to evaluate the association between treatment and OS. ResultsIn our cohort of 4,169 patients, multivariable analysis identified increasing age (RRR: 1.07, 95%CI, 1.06–1.08) and comorbidity burden (, as independent predictors of receiving immunotherapy. Treatment at an academic facility was associated with increased likelihood of combination treatment (RRR: 1.29, 95%CI, 1.01–1.65). After IPTW, we found that combination therapy (hazard ratio [HR]: 0.72; 95%CI, 0.62–0.83) was associated with improved survival compared to chemotherapy. ConclusionsPatients with older age and more comorbidities were more likely to receive immunotherapy than chemotherapy for first-line treatment of metastatic urothelial carcinoma of the bladder. Utilization of chemoimmunotherapy was observed to be higher in academic centers and was associated with improved survival compared to chemotherapy.

Abstract B006: GSTT2 modulates immune activation and impacts response to BCG immunotherapy in bladder cancer

Abstract Whilst M. bovis Bacillus Calmette-Guérin (BCG) therapy is the gold-standard for treating high risk non-muscle invasive bladder cancer (NMIBC), 30-40% of patients fail therapy, resulting in disease recurrence and progression. Loss of glutathione-S-transferase theta 2 (GSTT2) expression resulting from a promoter deletion, has been associated with modulation of intracellular ROS and BCG survival in BC cells. Retrospective analysis of NMIBC patients revealed that patients with the promoter deletion responded better to fewer instillations of BCG. To understand these responses, wildtype (WT) and GSTT2-knockout (KO) mice were implanted with MB49-PSA BC cells, either subcutaneously or orthotopically, and treated with weekly BCG instillations. Subcutaneous tumors were harvested after 3 BCG instillations for flow cytometry analysis, or after 4 BCG instillations for immunofluorescence (IF) imaging and gene expression analysis. Bladders (orthotopic model) were harvested after 4 BCG instillations for single-cell RNA sequencing. Splenocytes from WT and KO male mice were also stimulated in-vitro with TLR agonists, for 48h, to assess immune responses. In the subcutaneous model, in male mice, the cure rate was higher in WT mice (57%) compared to KO mice (0%). Flow cytometry analysis showed significantly higher numbers of tumor-associated CD8+ T-cells in BCG-treated KO males compared to WT males; however, IF imaging revealed higher penetration of CD8+ T-cells in tumors from WT males. In female mice, the cure rate was similar in WT (25%) and GSTT2-KO (38%) mice. IF imaging showed higher penetration of CD8+ T-cells in KO females. Quantitative real-time PCR (q-PCR) analysis of BCG-treated subcutaneous tumors from female mice revealed lower expression of the immune exhaustion markers, PD-L1 and CTLA-4, in KO mice compared to WT mice. Similarly, in the orthotopic model (female mice), q-PCR analysis of bladders also showed lower expression of PD-L1 and CTLA-4 in KO mice compared to WT mice. Single-cell RNA sequencing of bladders from BCG-treated mice revealed differences in the expression of genes such as Peak1, Hmga2, Scl2a1, Kras, Ahank, Gsta4, Mecom and Sh3gl2, of which Gsta4, Mecom, Sh3gl2, Hmga2 and Peak1 were validated using q-PCR. In-vitro analysis of splenocytes from male mice revealed higher levels of IL-6, IL-1β and IFN-α, after stimulation with TLR7/8 and TLR9 agonists, in KO splenocytes relative to WT splenocytes. Despite the higher levels of cytokine production by splenocytes and higher proportion of tumor-associated CD8+ T-cells in KO male mice, these mice fared poorly, which could be explained by immune exhaustion or lack of immune penetration into the tumor. In females, lower expression of immune exhaustion markers in KO mice did not yield better responses; however, single-cell data showed differences in expression of genes associated with pro- and anti-tumorigenic pathways, which could influence response. Thus, loss of GSTT2 may influence the response to BCG therapy, through modulating downstream signaling pathways and immune responses. Citation Format: Mugdha Vijay Patwardhan, Kane Toh, Edmund Chiong, Ratha Mahendran. GSTT2 modulates immune activation and impacts response to BCG immunotherapy in bladder cancer [abstract]. In: Proceedings of the AACR Special Conference on Bladder Cancer: Transforming the Field; 2024 May 17-20; Charlotte, NC. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(10_Suppl):Abstract nr B006.

Breaking Barriers: Modulation of Tumor Microenvironment to Enhance Bacillus Calmette–Guérin Immunotherapy of Bladder Cancer

Breaking Barriers: Modulation of Tumor Microenvironment to Enhance Bacillus Calmette–Guérin Immunotherapy of Bladder Cancer

Machine learning-based identification of a consensus immune-derived gene signature to improve head and neck squamous cell carcinoma therapy and outcome.

Head and neck squamous cell carcinoma (HNSCC), an extremely aggressive tumor, is often associated with poor outcomes. The standard anatomy-based tumor-node-metastasis staging system does not satisfy the requirements for screening treatment-sensitive patients. Thus, an ideal biomarker leading to precise screening and treatment of HNSCC is urgently needed. Ten machine learning algorithms-Lasso, Ridge, stepwise Cox, CoxBoost, elastic network (Enet), partial least squares regression for Cox (plsRcox), random survival forest (RSF), generalized boosted regression modelling (GBM), supervised principal components (SuperPC), and survival support vector machine (survival-SVM)-as well as 85 algorithm combinations were applied to construct and identify a consensus immune-derived gene signature (CIDGS). Based on the expression profiles of three cohorts comprising 719 patients with HNSCC, we identified 236 consensus prognostic genes, which were then filtered into a CIDGS, using the 10 machine learning algorithms and 85 algorithm combinations. The results of a study involving a training cohort, two testing cohorts, and a meta-cohort consistently demonstrated that CIDGS was capable of accurately predicting prognoses for HNSCC. Incorporation of several core clinical features and 51 previously reported signatures, enhanced the predictive capacity of the CIDGS to a level which was markedly superior to that of other signatures. Notably, patients with low CIDGS displayed fewer genomic alterations and higher immune cell infiltrate levels, as well as increased sensitivity to immunotherapy and other therapeutic agents, in addition to receiving better prognoses. The survival times of HNSCC patients with high CIDGS, in particular, were shorter. Moreover, CIDGS enabled accurate stratification of the response to immunotherapy and prognoses for bladder cancer. Niclosamide and ruxolitinib showed potential as therapeutic agents in HNSCC patients with high CIDGS. CIDGS may be used for stratifying risks as well as for predicting the outcome of patients with HNSCC in a clinical setting.

Reply to “Knowledge-map analysis and bladder cancer immunotherapy: Comment”

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Abstract 3924: Dual COX-2/sEH inhibition enhances immunotherapy and chemotherapy to induce bladder cancer regression

Abstract Unresolved inflammation plays a critical role in bladder cancer initiation and progression. Chemotherapy (e.g. gemcitabine and cisplatin), the standard of care for advanced bladder cancer, disrupts inflammation resolution and is only partially effective in preventing tumor recurrence after treatment. Immunotherapy has emerged as a potential treatment option for bladder cancer patients, however it is ineffective in 80% of patients and can induce a pro-inflammatory cytokine storm. Therefore, there is a critical unmet medical need to improve chemotherapy and immunotherapy in bladder cancer. To address this, we have developed dual COX-2/sEH inhibitors (e.g. PTUPB). This lead compound is representative of a newly patented class of “dual acting” molecules that target two important enzymes in the arachidonic acid cascade with nanomolar binding affinity: cyclooxygenase-2 (COX-2) and soluble epoxide hydrolase (sEH). Oral dosing of PTUPB is well-tolerated in animal models. Since chemotherapy and immunotherapy both induce tumor-promoting inflammation via an eicosanoid/cytokine storm, we hypothesized that dual COX-2/sEH inhibition would enhance immunotherapy in experimental bladder cancer via anti-inflammatory mechanisms. When syngeneic (MB49) bladder tumors reached ~200 mm3 in immunocompetent mice, treatment was initiated with PTUPB, anti-CTLA-4, anti-PD1, gemcitabine & cisplatin, or various combinations thereof. Here we demonstrate that chemotherapy and/or immunotherapy stimulated sEH and COX-2 expression in tumor tissue which was counter-regulated by PTUPB. While monotherapy treatment with PTUPB, gemcitabine & cisplatin, anti-PD1 or anti-CTLA-4 suppressed tumor growth compared to control, the tumors escaped single treatments by the end of the study. Remarkably, dual COX-2/sEH inhibition in combination with chemotherapy (gemcitabine and cisplatin) and/or immune checkpoint blockade (anti-CTLA-4 or anti-PD1) induced sustained tumor regression via synergistic anti-tumor activities. Chemotherapy and/or immunotherapy induced the expression of ER stress response genes (e.g. BiP and CHOP) and angiogenic factors (e.g. EGF and VEGF-C) in bladder cancer tissue, which were repressed by PTUPB. PTUPB also prevented chemotherapy-induced toxicity and prolonged survival in an orthotopic MB49 tumor model. Taken together, our results demonstrate dual COX-2/sEH inhibition as a novel therapeutic approach to enhance immunotherapy in bladder cancer without overt toxicity. Citation Format: Kimberly Lupita Vazquez, Eva Rothenberger, Weicang Wang, Sung Hee Hwang, Diane R. Bielenberg, Bruce D. Hammock, Dipak Panigrahy. Dual COX-2/sEH inhibition enhances immunotherapy and chemotherapy to induce bladder cancer regression [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 3924.

Abstract 7611: Circulating blood RNAseq detects activation of immune system and predicts response to immunotherapy in bladder cancer

Abstract Background: Anti-PD(L)1 immunotherapies have revolutionized oncology by getting durable tumor responses in advanced cancers. Nevertheless, currently approved biomarkers (PD-L1, MSI, TMB) have suboptimal positive/negative predictive values for tumor responses and survival. Aims & Methods: We sought to assess the value of gene expression on circulating blood to predict responses to immunotherapy. We performed total paired-end RNAseq at 20-million reads and analyzed differential gene expressions according to cancer outcomes on baseline frozen whole blood from a cohort of 186 patients prospectively enrolled in the IOPREDI study, a French ancillary cohort of the international multi-center STRONG Phase III trial testing the anti-PD-L1 durvalumab in advanced bladder cancers (NCT03084471). Results: One hundred and sixty four out of the 186 baseline samples generated valuable blood transcriptome data. Thanks to mathematical corrections, all samples were normalized, and the analysis was possible without introducing any bias despite the sample origin from 16 enrolling sites. We first observed that patients with a complete or partial response or a stable disease overexpressed 19 genes, 5 of which are markers of immune cell activation: CXCL8, EOMES, GBP5, GZMA, and GZMH. Conversely, patients with a progressive disease overexpressed 11 genes including CD177, a marker of neutrophil cells, recently associated to immunotherapy resistance. Next, patients with a PFS > 3 months showed an overexpression of 25 genes associated with immune cells activation such as CD8A, CXCL8, EOMES, FASLG, GZM genes, whereas patients with a shorter PFS highly expressed genes of immune cell inhibition: CD177, CD300LG, CD99, and CST7. Finally, patients with an OS > 7 months had also a higher expression of the same previous genes completed with 6 other genes, still indicators of immunity activation (CD3D, CD3E, CD3G, GZMM, IL2RB, and PTGDS). Conclusion: Total RNAseq from RNA directly extracted from whole blood can detect baseline activation of T-cells in patients who will subsequently benefit from anti-PD-L1 immunotherapy. This work shows that RNAseq can detect an immune activation state in circulating blood without specific cell selection. This technique could be used in clinical trials to select for patients prone to benefit from cancer immunotherapies. Citation Format: Corentin Richard, Sandy Chevrier, Aurélien Marabelle, Romain Boidot. Circulating blood RNAseq detects activation of immune system and predicts response to immunotherapy in bladder cancer [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 7611.

Abstract 2515: Cation transport gene signature predicts overall and progression free survival in bacillus-calmette-guerrin-treated non-muscle invasive bladder cancer in The Cancer Genome Atlas

Abstract Introduction: Bacillus Calmette-Guerrin (BCG) is a commonly used immunotherapy for non-muscle invasive bladder cancer (NMIBC), and there is limited understanding of its in vivo mechanisms. In vitro studies on patient-derived tumor cells suggest the role of calcium signaling as an effector of immune response. We aimed to elucidate further understanding of important signaling pathways implicated in BCG response using gene expression data. The goal is to identify a gene signature for predicting BCG response and survival post-BCG therapy. Methods: Gene expression data for the BCG treated NMIBC cohort was downloaded from TCGA. Individual gene expression data analyzed with univariate cox proportional hazard models versus overall survival (OS) and progression-free survival (PFS) independently. The survival outputs were then fed into String DB network analysis. Overlapping genes in the 2 networks were combined into a Z- sum composite score for survival prediction. Correlation analysis was performed for the overlapping genes. Results: Based on the selection criteria, 35 patients were available for the study. In OS analysis a total of 622 genes were filtered based on HR>2 or <0.5 with adjusted-p <0.05. Network analysis of those significant genes demonstrated significant calcium ion transmembrane transport upregulation in patients with better OS, showing a subnetwork of 16 genes. In the PFS analysis, 500 genes with HR>2 or <0.5 with adjusted-p<0.05 were subjected to network analysis. The main cluster was enriched in cation transmembrane transport upregulated in patients with better PFS, and consisted of a subnetwork of 39 genes. There were 295 shared genes between the genes associated with improved OS and PFS gene sets. A composite score constructed from these 295 genes revealed an optimized cutoff of the lower 70th percentile of expression versus the upper 30th percentile of normalized summed gene expression (HR = 0.192, p = 0.00048 for OS; and HR = 0.171, p < 0.0001 for PFS). Internal correlation reveals 3 sub-clusters of gene expression within our data set. Conclusion: Similar to in vitro studies we report increased cation transport expression in vivo, that was significantly associated with improved OS after BCG immunotherapy. Our composite gene signature score was able to identify a poor survival subgroup—about 1/3 of patients—with 5-fold worse PFS and OS following BCG therapy. Increasing calcium signaling may potentiate BCG response in currently poor BCG-responders. Citation Format: Lynn Kim Hoang Tran, Seth P. Lerner. Cation transport gene signature predicts overall and progression free survival in bacillus-calmette-guerrin-treated non-muscle invasive bladder cancer in The Cancer Genome Atlas [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 2515.

Systemic versus localized Bacillus Calmette Guérin immunotherapy of bladder cancer promotes an anti-tumoral microenvironment: Novel role of trained immunity.

Treatment for higher-risk non-muscle invasive bladder cancer (NMIBC) involves intravesical immunotherapy with Bacillus Calmette Guérin (BCG); however, disease recurrence and progression occur frequently. Systemic immunity is critical for successful cancer immunotherapy; thus, recurrence of NMIBC may be due to suboptimal systemic activation of anti-tumor immunity after local immunotherapy. We previously reported that systemically acquired trained immunity (a form of innate immune memory) in circulating monocytes is associated with increased time-to-recurrence in patients with NMIBC treated with BCG. Herein, we used a mouse model of NMIBC to compare the effects of intravesical versus intravenous (systemic) BCG immunotherapy on the local and peripheral immune microenvironments. We also assessed whether BCG-induced trained immunity modulates anti-tumor immune responses. Compared with intravesical BCG, which led to a tumor-promoting immune microenvironment, intravenous BCG resulted in an anti-tumoral bladder microenvironment characterized by increased proportions of cytotoxic T lymphocytes (CTLs), and decreased proportions of myeloid-derived suppressor cells. Polarization toward anti-tumoral immunity occurred in draining lymph nodes, spleen, and bone marrow following intravenous versus intravesical BCG treatment. Pre-treatment with intravesical BCG was associated with increased rate of tumor growth compared with intravenous BCG pre-treatment. Trained immunity contributed to remodeling of the tumor immune microenvironment, as co-instillation of BCG-trained macrophages with ovalbumin-expressing bladder tumor cells increased the proportion of tumor-specific CTLs. Furthermore, BCG-trained dendritic cells exhibited enhanced antigen uptake and presentation and promoted CTL proliferation. Our data support the concept that systemic immune activation promotes anti-tumor responses, and that BCG-induced trained immunity is important in driving anti-tumor adaptive immunity.

Multi-cohort validation of Ascore: an anoikis-based prognostic signature for predicting disease progression and immunotherapy response in bladder cancer

Bladder cancer ranks as the 10th most common cancer worldwide, with deteriorating prognosis as the disease advances. While immune checkpoint inhibitors (ICIs) have shown promise in clinical therapy in both operable and advanced bladder cancer, identifying patients who will respond is challenging. Anoikis, a specialized form of cell death that occurs when cells detach from the extracellular matrix, is closely linked to tumor progression. Here, we aimed to explore the anoikis-based biomarkers for bladder cancer prognosis and immunotherapeutic decisions. Through consensus clustering, we categorized patients from the TCGA-BLCA cohort into two clusters based on anoikis-related genes (ARGs). Significant differences in survival outcome, clinical features, tumor immune environment (TIME), and potential ICIs response were observed between clusters. We then formulated a four-gene signature, termed "Ascore", to encapsulate this gene expression pattern. The Ascore was found to be closely associated with survival outcome and served as an independent prognosticator in both the TCGA-BLCA cohort and the IMvigor210 cohort. It also demonstrated superior predictive capacity (AUC = 0.717) for bladder cancer immunotherapy response compared to biomarkers like TMB and PD-L1. Finally, we evaluated Ascore’s independent prognostic performance as a non-invasive biomarker in our clinical cohort (Gulou-Cohort1) using circulating tumor cells detection, achieving an AUC of 0.803. Another clinical cohort (Gulou-Cohort2) consisted of 40 patients undergoing neoadjuvant anti-PD-1 treatment was also examined. Immunohistochemistry of Ascore in these patients revealed its correlation with the pathological response to bladder cancer immunotherapy (P = 0.004). Impressively, Ascore (AUC = 0.913) surpassed PD-L1 (AUC = 0.662) in forecasting immunotherapy response and indicated better net benefit. In conclusion, our study introduces Ascore as a novel, robust prognostic biomarker for bladder cancer, offering a new tool for enhancing immunotherapy decisions and contributing to the tailored treatment approaches in this field.

Bibliometric analysis of immunotherapy for bladder cancer: A correspondence

Bibliometric analysis of immunotherapy for bladder cancer: A correspondence

Quality analysis of BCG vaccine for bladder cancer immunotherapy using Shewhart control charts

SCIENTIFIC RELEVANCE. The complexity of standardising immunobiologicals, including bacillus Calmette–Guérin (BCG) vaccines, requires improving the approach to statistical analysis of consistency in their quality indicators. Quality analysis tools include Shewhart control charts, which can be used for real-time and retrospective identification and analysis of trends in the processes of interest.AIM. This study aimed to analyse quality indicators of a BCG vaccine for bladder cancer immuno­therapy using Shewhart control charts to monitor consistency in manufacturing processes and quality testing of vaccines.MATERIALS AND METHODS. The study involved a retrospective analysis of the bacterial concentration, dispersity index, loss-on-drying, and potency data obtained during the quality control of Imuron-vac, a BCG vaccine for bladder cancer immunotherapy, in the past 5 years. As part of the statistical data analysis, the authors calculated the means, investigated the corre­lation between the test results provided by the manufacturer and the quality control results obtained at the testing centre (TC), and checked the distributions for normality. Individuals and moving range control charts were plotted and analysed for each quality indicator.RESULTS. The correlation coefficients (r) ranged from 0.34 to 0.70 for the data submitted by the manufacturer and the data obtained at the TC. Pearson’s goodness-of-fit test showed that the distribution of the quality control results obtained at the TC was normal, with χ2 = 14.03 for the dispersity index values and χ2 = 4.93 for the loss-on-drying measurements. Shewhart control charts for loss-on-drying and potency data suggested that the production process and/or the testing process deviated from the state of statistical control. Consistency monitoring of the bacterial concentration and the dispersity index showed that the state of statistical control was achieved. The authors established control limits for further real-time monitoring of the dispersity index consistency.CONCLUSIONS. Shewhart control charts are applicable to the quality control of BCG vaccines for bladder cancer immunotherapy at the TC. Continuous monitoring of consistency in quality indicators is needed to improve the standardisation of vaccine production processes and testing methods, as this standardisation will result in high product quality.

Analysis of neoadjuvant immunotherapy and chemotherapy for muscle-invasive bladder cancer in a national registry.

Analysis of neoadjuvant immunotherapy and chemotherapy for muscle-invasive bladder cancer in a national registry.

POLQ identifies a better response subset to immunotherapy in muscle-invasive bladder cancer with high PD-L1.

Though programmed cell death-ligand 1 (PD-L1) has been used in predicting the efficacy of immune checkpoint blockade (ICB), it is insufficient as a single biomarker. As a key effector of an intrinsically mutagenic microhomology-mediated end joining (MMEJ) pathway, DNA polymerase theta (POLQ) was overexpressed in various malignancies, whose expression might have an influence on genomic stability, therefore altering the sensitivity to chemotherapy and immunotherapy. A total of 1304 patients with muscle-invasive bladder cancer (MIBC) from six independent cohorts were included in this study. The Zhongshan Hospital (ZSHS) cohort (n = 134), The Cancer Genome Atlas (TCGA) cohort (n = 391), and the Neo-cohort (n = 148) were included for the investigation of chemotherapeutic response. The IMvigor210 cohort (n = 234) and the UNC-108 cohort (n = 89) were used for the assessment of immunotherapeutic response. In addition, the relationship between POLQ and the immune microenvironment was assessed, and GSE32894 (n = 308) was used only for the evaluation of the immune microenvironment. We identified POLQhigh PD-L1high patients could benefit more from immunotherapy and platinum-based chemotherapy. Further analysis revealed that high POLQ expression was linked to chromosome instability and higher tumor mutational burden (TMB), which might elicit the production of neoantigens. Further, high POLQ expression was associated with an active tumor immune microenvironment with abundant infiltration of immune effector cells and molecules. The study demonstrated that high POLQ expression was correlated with chromosome instability and antitumor immune microenvironment in MIBC, and the combination of POLQ and PD-L1 could be used as a superior companion biomarker for predicting the efficacy of immunotherapy.

Abstract A020: An early determination of patients eligibility for a bladder cancer immunotherapy using a data science approach

Abstract Advanced bladder cancer patients have limited treatment options and, in most cases, limited overall survival. The first clinical trial with adoptive T cell therapy (ACT) that uses bladder cancer patients’ autologous tumor-infiltrating T lymphocytes (TILs) has been recently opened at Moffitt. To help identify which patients could benefit from ACT-TIL we are developing a machine learning predictor that takes advantage of tumor evolution data, such as tumor stages and grades routinely collected in clinic. The first phase of ACT-TIL involves resection of the primary tumor and the growth of TILs followed by a rapid expansion protocol of the reactive T cells. It may take up to 4-5 weeks before bladder cancer patients can get their own expanded TILs. Our predictor aims to determine whether the tumor isolated from a patient will grow TILs. In this work, we present a machine learning predictor of expansion of tumor infiltrating lymphocytes (ML-PETIL) that uses retrospectively collected patient tumor data with either demographic, clinical, or biological sample-related features. The proposed method consists of a suite of algorithms to identify a robust set of predictive features (feature selection algorithms) that are used to discriminate the dataset into two classes-that with successful TIL expansion or with no TIL growth (classification algorithms). Our predictor includes metrics which are used to address data imbalance and handle small to medium-sized datasets that contain features of varying datatypes and magnitudes. The ML-PETIL protocol offers an early determination of patients who would be eligible for ACT-TIL therapy. Citation Format: Kayode D. Olumoyin, Ahmet M. Aydin, Brittany L Bunch, Shari Pilon-Thomas, Michael Poch, Katarzyna A. Rejniak. An early determination of patients eligibility for a bladder cancer immunotherapy using a data science approach [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Translating Cancer Evolution and Data Science: The Next Frontier; 2023 Dec 3-6; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(3 Suppl_2):Abstract nr A020.

Bladder Cancer: Immunotherapy and Pelvic Lymph Node Dissection.

Bladder cancer, a common malignancy of the urinary system, is routinely treated with radiation, chemotherapy, and surgical excision. However, these strategies have inherent limitations and may also result in various side effects. Immunotherapy has garnered considerable attention in recent years as a novel therapeutic approach. It harnesses and activates the patient's immune system to recognize and eliminate cancer cells, which not only prolongs therapeutic efficacy but also minimizes the toxic side effects. Several immune checkpoint inhibitors and cancer vaccines have been developed for the treatment of bladder cancer. Whereas blocking immune checkpoints on the surface of tumor cells augments the effect of immune cells, immunization with tumor-specific antigens can elicit the production of anti-tumor immune effector cells. However, there are several challenges in applying immunotherapy against bladder cancer. For instance, the efficacy of immunotherapy varies considerably across individual patients, and only a small percentage of cancer patients are responsive. Therefore, it is crucial to identify biomarkers that can predict the efficacy of immunotherapy. Pelvic lymph nodes are routinely dissected from bladder cancer patients during surgical intervention in order to remove any metastatic tumor cells. However, some studies indicate that pelvic lymph node dissection may reduce the efficacy of immunotherapy by damaging the immune cells. Therefore, the decision to undertake pelvic lymph node removal should be incumbent on the clinical characteristics of individual patients. Thus, although immunotherapy has the advantages of lower toxic side effects and long-lasting efficacy, its application in bladder cancer still faces challenges, such as the lack of predictive biomarkers and the effects of pelvic lymph node dissection. Further research is needed to explore these issues in order to improve the efficacy of immunotherapy for bladder cancer.

Targeting STAT3 in tumor-associated antigen-presenting cells as a strategy for kidney and bladder cancer immunotherapy.

Immune checkpoint blockade (ICB) improved clinical outcomes in renal and bladder cancer patients, but the response rates remain limited especially in metastatic disease. While STAT3 transcription factor is well-known master regulator of tumor immune evasion, little is known about the role of STAT3 in the resistance of renal or bladder cancers to immunotherapy. To better understand immune alterations associated with ICB resistance, we assessed blood biomarkers in renal cancer patients classified as responders or non-responders to first line nivolumab/ipilimumab immunotherapy. We observed that non-responders showed elevated levels of proinflammatory mediators, such as IL-1RA, IL-6, IL-8 and to lesser extent IL-10, which are associated with STAT3 activation and tumor immunosuppression. In addition, we found STAT3 activation primarily in circulating myeloid immune cells such as tolerogenic MDSCs. To assess whether STAT3 inhibition within these cell subsets can promote antitumor immune responses and/or enhance sensitivity to ICB in vivo, we used an original antisense oligonucleotide (ASO) strategy for myeloid-cell selective STAT3 knockdown (CpG-STAT3ASO). Our results in syngeneic models of renal and bladder cancers in mice demonstrated potent antitumor activity of CpG-STAT3ASO alone in contrast to PD1 blockade alone in both models. The CpG-STAT3ASO/anti-PD1 combination improved therapeutic efficacy especially against bladder tumors. Therapeutic efficacy correlated with activation of dendritic cells (DCs) and M1 macrophages in the tumor microenvironment, reduced percentages of regulatory T cells (Tregs) and the expansion of CD8 T cells in both tumor models. Our study underscores the potential of using myeloid-cell targeted CpG-STAT3 inhibitors for genitourinary cancer therapy to disrupt tolerogenic signaling, restore immune cell activity and sensitivity to immune checkpoint inhibitors and/or T cell-based immunotherapies.

Nanoreactor based on single-atom nanoenzymes promotes ferroptosis for cancer immunotherapy

Immunotherapy is a promising mainstream approach in anti-tumor therapy. It boasts advantages such as durable responses and lower side effects. However, there are still some limitations to be addressed. Current cancer immunotherapy has shown low response rates due to inadequate immunogenicity of certain tumor cells. To address these challenges, an acid-specific nanoreactor was developed, designed to induce immunogenicity by triggering ferroptosis in tumor cells. The nanoreactor integrates glucose oxidase (GOx) with a single-atom nanoenzyme (SAE), which exhibits high peroxidase (POD)-like activity in the acidic tumor microenvironment (TME). This specific acid-sensitivity transforms endogenous hydrogen peroxide (H2O2) into cytotoxic hydroxyl radicals (•OH). GOx enhances the POD-like SAE activity in the nanoreactor by metabolizing glucose in tumor cells, producing gluconic acid and H2O2. This nanoreactor induces high levels of oxidative stress within tumor cells through the synergistic action of SAE and GOx, leading to depletion of GSH and subsequently triggering ferroptosis. The resulting nanoreactor-induced ferroptosis leads to immunogenic cell death (ICD) and significantly recruits T lymphocyte infiltration in tumor tissues. This study was designed with the concept of triggering ferroptosis-dependent ICD mechanism in bladder cancer cells, and developed an acid-specific nanoreactor to enhance the immunotherapy efficacy for bladder cancer, which introduces a novel approach for immunotherapy of bladder cancer.

Electrocardiographic characteristics of bladder cancer patients receiving preoperative chemotherapy combined with immunotherapy.

Patients treated with preoperative chemotherapy and immunotherapy for bladder cancer may be at increased risk of cardiotoxicity and electrophysiological abnormalities. This study aimed to analyze their electrocardiographic (ECG) alterations. Patients with bladder cancer who were hospitalized and receiving tislelizumab plus nab-paclitaxel (TnP) were enrolled prospectively. ECG, cardiac biomarkers, and echocardiography were performed at baseline and the end of TnP. A total of 60 patients (76.7% males), including 30 muscle-invasive and 30 non-muscle-invasive bladder cancer, received three or four cycles of TnP, respectively. Hypertension was the commonest comorbidity (41.7%), and 25 patients (41.7%) were prescribed cardiovascular drugs. In comparison with baseline characteristics, cardiac troponin I (cTnI) and N-terminal pro-brain natriuretic peptide (NT-proBNP) were within normal ranges after TnP. However, echocardiographic parameter of left ventricular ejection fraction slightly decreased after TnP (62.81 ± 3.81% to 61.10 ± 4.37%, p = .011). The incidence of abnormal ECG increased from 65.0% at baseline to 76.7%, of which only a higher prevalence of fragmented QRS (fQRS) was observed (33.3% to 50.0%, p = .013; mainly in inferior leads). ECG parameters of QT dispersion (QTd) were prolonged significantly after the regimen (39.50 ± 11.37 to 44.20 ± 15.85 ms, p = .019). In bladder cancer patients receiving preoperative chemotherapy combined with immunotherapy, the main ECG abnormality was fQRS and QTd, with relatively normal cardiac biomarkers and echocardiographic parameters. Regular ECG screening should be carried out carefully to detect potential cardiotoxicity in the long-term follow-up.