BACKGROUND AND OBJECTIVE:To determine whether the existing European Organisation for Research and Treatment of Cancer (EORTC) QLQ-NMIBC24 and QLQ-BLM30 modules remain fit for purpose and to assess health-related quality of life (HRQoL) issues relevant for patients with metastatic bladder cancer (mBC). METHODS:Following the EORTC Quality of Life Group (QLG) guidelines, phase 1 included an expert meeting, five systematic reviews, and interviews with 14 healthcare professionals (HCPs) and 48 bladder cancer (BC) patients across six countries. HRQoL issues were identified through the literature and rated by HCPs and patients for relevance and importance. Decision rules guided selection for phase 2. In phase 2, selected issues were translated into items using the EORTC Item Library and expert consensus. KEY FINDINGS AND LIMITATIONS:A total of 443 articles and 62 interviews yielded 150 unique HRQoL issues; 40 issues and two branching questions were retained for the provisional module. Seventeen new items were added beyond the existing EORTC BC modules, while 11 existing items were excluded. Issues specific to mBC and newer treatments, such as immunotherapy, were identified. Limitations include potential selection bias due to purposive sampling and small sample sizes per treatment subgroup. CONCLUSIONS AND CLINICAL IMPLICATIONS:This phase 1-2 study supports the need to update the existing EORTC BC modules and incorporate additional items relevant to all disease stages, including mBC. A single, flexible module with branching logic may improve clinical applicability across patient subgroups. The provisional module will undergo international pretesting and psychometric evaluation in phase 3.

The intratumoral microbiome has recently emerged as a potential hallmark of cancer, with implications for response or resistance to therapy. Bacteria can either promote or inhibit cancer growth. However, intratumoral bacteria can also be engineered using synthetic biology to remodel the tumor microenvironment. Here, we engineered the probiotic bacterium Escherichia coli Nissle 1917 (EcN) to express the human chemokine CXCL13 (C-X-C motif chemokine ligand 13), a critical component of germinal center (GC) formation. Antibody affinity maturation and class switching are fundamental aspects of adaptive immune response. Both occur primarily in the GCs of secondary lymphoid organs for defense against pathogens. Immune checkpoint blockade (ICB) efficacy is primarily driven by T cells; however, recent studies in mice and humans have shown that humoral immune responses act as critical partners for ICB-mediated antitumor activity. Using orthotopic models of bladder cancer, intravesically delivered engineered CXCL13-expressing EcN colonized bladder tumors and elicited GC responses in bladder tumor-draining lymph nodes after intravesical delivery. When combined with programmed cell death protein 1 (PD-1) blockade, engineered EcN improved antitumor activity in two aggressive, fast-growing, and immunologically cold orthotopic mouse models of bladder cancer. Mechanistically, this antitumor effect was dependent on the presence of CD8+ T cells and CD4+ T follicular helper cells; combination therapy increased tumor-specific antibody responses and promoted long-term survival and protective immunity upon tumor rechallenge. Thus, we demonstrate that synthetically engineered CXCL13-expressing EcN can enhance the efficacy of PD-1 checkpoint blockade immunotherapy by amplifying tumor-specific humoral immunity.

BACKGROUND:In the tumor microenvironment, immune, stromal and cancer cells interact. They communicate in paracrine and autocrine manners via secreted cytokines, altering anti-tumor immunity. The pattern consisting of the relative secretions of these proteins is referred-to as the "immune secretome". To date, the immune secretome of human cancers remains largely unexplored. Herein, we describe the dominant cytokines released by human tumors. METHODS:We analyzed multiple prospectively collected samples of tumoral and peritumoral tissues. After surgical resection, we promptly titered 40 cytokines released by the samples. RESULTS:Macrophage migration Inhibitory Factor (MIF), Chemokine (C-C motif) ligand 21 (CCL21) and Interleukin 16 (IL-16) were the most strongly detected cytokines in the human tumor secretome. Surprisingly, GM-CSF, IL-10 and IL-2 were undetectable or measured at low levels. Moreover, we observed significant differences in cytokine secretions across different tumor histotypes: while liver metastases, head and neck carcinomas and thyroid tumors secreted large amounts of cytokines (103-105 pg/ gram of tissue across most analytes), the other histotypes secreted at least 10-100 times less of each cytokine per gram of tissue. The liver metastases secreted particularly high amounts of CCL21, IL-16, CXCL10, CCL24 and CCL15, but had low CD45 + cell infiltrates compared to primary tumors. We also found that tumor tissues secreted significantly higher quantities of CXCL1 and CXCL9 than surrounding healthy tissues. CONCLUSIONS:Tumor histology is not associated with a specific secretome, but subgroups of patients share similar secretome profiles independently from their cancer type. This finding supports the development of biology-oriented tumor-agnostic immunotherapy strategies. As the secretome interacts with tumor-infiltrating T-cells, the cytokines identified within it become potential therapeutic targets for personalized cancer immunotherapy.

Background and objective There is no standardized regimen for follow-up after radical cystectomy (RC) for bladder cancer (BC). To address this gap, we conducted a multicenter study involving urologist members from the European Association of Urology (EAU) bladder cancer guideline panels. Our objective was to identify consistent post-RC follow-up strategies and develop a practice-based framework based on expert opinion. Methods We surveyed 27 urologist members of the EAU guideline panels for non–muscle-invasive bladder cancer and muscle-invasive and metastatic bladder cancer using a pre-tested questionnaire with dichotomous responses. The survey inquired about follow-up strategies after RC and the use of risk-adapted strategies. Consistency was defined as >75% affirmative responses for follow-up practices commencing 3 mo after RC. Descriptive statistics were used for analysis. Key findings and limitations We received responses from 96% of the panel members, who provided data from 21 European hospitals. Risk-adapted follow-up is used in 53% of hospitals, with uniform criteria for high-risk (at least ≥pT3 or pN+) and low-risk ([y]pT0/a/1N0) cases. In the absence of agreement for risk-based follow up, a non-risk-adapted framework for follow-up was developed. Higher conformity was observed within the initial 3 yr, followed by a decline in subsequent follow-up. Follow-up was most frequent during the first year, including patient assessments, physical examinations, and laboratory tests. Computed tomography of the chest and abdomen/pelvis was the most common imaging modality, initially at least biannually, and then annually from years 2 to 5. There was a lack of consistency for continuing follow-up beyond 10 yr after RC. Conclusions and clinical implications This practice-based post-RC follow-up framework developed by EAU bladder cancer experts may serve as a valuable guide for urologists in the absence of prospective randomized studies. Patient summary We asked urologists from the EAU bladder cancer guideline panels about their patient follow-up after surgical removal of the bladder for bladder cancer. We found that although urologists have varying approaches, there are also common follow-up practices across the panel. We created a practical follow-up framework that could be useful for urologists in their day-to-day practice.

Treatment and dose modifications in patients according to their DCB status.

The observation that bacteria colonize tumors, but not normal organs, after intravenous administration has spurred new interest in bacteria as tumor-targeted delivery systems. Since bacteria replicate within the tumor site, intravenous administration of bacteria leads to specific expression of peptide-based therapeutics at the tumor site. However, most prior bacterial cancer therapies have relied on this natural tumor tropism since the mechanisms behind this tumor-specificity are poorly understood. Additionally, clinical trials with bacterial therapeutics have shown lower tumor colonization rates in humans, which limits their translatability. This study used a pooled barcode library approach to screen the E. coli genome for pathways which may influence tumor colonization. We found specific pathways, include glucose catabolism, which promote E. coli tumor colonization, and non-glucose catabolic pathways which inhibit E. coli tumor colonization. KO of these inhibitory metabolic pathways increase E. coli glucose uptake and utilization. This was associated with increased tumor colonization as validated by competitive infection experiments, which was further enhanced when glucose was introduced into the tumor microenvironment. Together, these results support a major role for E. coli glucose catabolism in defining bacterial tumor tropism and suggest a promising method by which bacterial tumor tropism can be fine-tuned. Noah Chen, Mathieu Rouanne, Zaofeng Yang, Tal Danino, Nicholas Arpaia. Engineering tumor-avid bacteria as anticancer immunotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 959.

Additional significant differences in circulating chemokines.

Cytotoxic response in tumor biopsies of patients with No DCB upon combination treatment.

Further descriptions of tumor immune infiltrates.

Adverse events emergent during treatment and related to experimental drugs by the investigators according to CTCAE Version 4.03.

Gating strategy for flow cytometry analyses of T cells infiltration on tumor biopsies.

Immuno-Histo-Chemistry scoring of baseline tumor biopsies according to their DCB status.

Differential Gene Expression (DGE) of baseline tumor biopsies between patients with DCB and No DCB.

Immune checkpoint blockade (ICB) have revolutionized medical oncology, with the occurrence of cures in situations regarded as palliative. However, most patients develop resistance and the selection of responders remains difficult. The search for predictive biomarkers for immunotherapy has revealed that tertiary lymphoid structures (TLS) in the tumor microenvironment (TME) may be indicative of treatment effectiveness, since they may reflect the presence of an immune infiltrate with an anti-tumor effect. A better understanding of TLS neogenesis would help to establish anti-tumor strategies. For this purpose, this review outlines therapeutic modalities that could increase the immunogenicity and antigenicity of tumor cells, in particular immunogenic cell death (ICD), to turn "cold" tumors in "hot" tumors through interventions that promote the development of TLS. Second, the review describes the therapeutic strategies to enhance immune response against tumors, especially the resources to induce TLS, to strengthen cytotoxic T response / decrease Tregs and to promote the homing of immune cells. Future strategies should focus on a synergistic tactic combining both the enhancement of cancer cell immunogenicity and the stimulation of the immune system. PROVENANCE AND PEER REVIEW: Review commissioned by Aurelien Marabelle; externally peer reviewed.

Baseline characteristics of the patients treated.

Production of amphiregulin (Areg) by regulatory T (Treg) cells promotes repair after acute tissue injury. Here, we examined the function of Treg cells in non-alcoholic steatohepatitis (NASH), a setting of chronic liver injury. Areg-producing Treg cells were enriched in the livers of mice and humans with NASH. Deletion of Areg in Treg cells, but not in myeloid cells, reduced NASH-induced liver fibrosis. Chronic liver damage induced transcriptional changes associated with Treg cell activation. Mechanistically, Treg cell-derived Areg activated pro-fibrotic transcriptional programs in hepatic stellate cells via epidermal growth factor receptor (EGFR) signaling. Deletion of Areg in Treg cells protected mice from NASH-dependent glucose intolerance, which also was dependent on EGFR signaling on hepatic stellate cells. Areg from Treg cells promoted hepatocyte gluconeogenesis through hepatocyte detection of hepatic stellate cell-derived interleukin-6. Our findings reveal a maladaptive role for Treg cell-mediated tissue repair functions in chronic liver disease and link liver damage to NASH-dependent glucose intolerance.

Background and objective Failure rates after first-line treatment of localized prostate cancer (PCa) treatment remain high; therefore, it is essential to improve the selection and identification of at-risk patients to reduce mortality. The aim of the ANDROCAN study was to evaluate the biochemical recurrence (BCR) in patients with localized PCa treated by total prostatectomy at 5 yr after surgery, according to their presurgery gonadal status. Methods A prospective cohort study was conducted including 1318 patients undergoing total prostatectomy for localized PCa with a 5-yr postoperative follow-up. Clinical and hormonal data (assays of total testosterone [TT], bioavailable testosterone [BT], dihydrotestosterone, estrone, and estradiol were performed by gas chromatography/mass spectrometry) as well as metabolic syndrome parameters were collected at baseline before surgery. Pathological data (predominant Gleason grade 4 and stage) were collected and cross-referenced centrally. Factors associated with BCR were assessed by a multivariate analysis, and BCR-free survival was assessed by a Kaplan-Meier analysis. Key findings and limitations Among the 1318 patients, 237 had BCR of PCa. Considering demographic characteristics, populations with and without BCR were similar. However, patients with BCR had cancers with a higher Gleason score (p = 0.0001) and higher prostate-specific antigen (PSA) values (p = 0.0005) at baseline. Gleason score, pT >3a, and PSA level at baseline were positively correlated with BCR (p < 0.0001, p < 0.0001, and p = 0.0048, respectively), while BT and TT levels were not associated with BCR. This study includes patients with varying clinical characteristics, such as cancer history and metabolic syndrome, introducing variability that makes it challenging to isolate the specific effects of gonadal status on BCR. Another limitation is the lack of evaluation of long-term BCR beyond 5 yr, potentially overlooking recurrences that occur between 5 and 15 yr after surgery. This could lead to an underestimation of the actual long-term recurrence rates. Conclusions and clinical implications Overall, PSA levels, high Gleason score, and pT >3a are associated with a greater likelihood of disease recurrence following initial treatment and could serve as important prognostic indicators for predicting the risk of BCR. In this prospective study, biochemical hypogonadism was not associated with a higher occurrence of BCR within 5 yr of prostatectomy. The biological gonadal status of preoperative patients could potentially be useful for therapeutic decisions but does not provide an indication for the oncological follow-up. Patient summary Five-year follow up of patients after surgery showed that there is no association between hypogonadism (low levels of total testosterone and bioavailable testosterone) and cancer recurrence. However, cancer recurrence seems to be more associated with aggressiveness of cancer at the time of detection.

The intratumoral microbiome has recently emerged as a new hallmark of cancer, with implications for response or resistance to therapy. While bacteria can either promote or inhibit cancer growth, intratumoral bacteria can also be engineered using synthetic biology to remodel the tumor microenvironment. Here, we engineered the probiotic bacterium E. coli Nissle 1917 (EcN) to express the human chemokine CXCL13, a critical component of germinal center (GC) formation. The GC reaction is a fundamental aspect of adaptive immunity by which antibody affinity develops in secondary lymphoid organs for defense against pathogens. Using orthotopic models of bladder cancer, engineered CXCL13-expressing EcN colonized bladder tumors and elicited GC responses in bladder tumor-draining lymph nodes after intravesical delivery. Furthermore, when combined with PD-1 blockade, engineered EcN amplified the antitumor antibody response and promoted long-term survival and protective immunity upon tumor rechallenge. Thus, we demonstrate that synthetically engineered CXCL13-expressing EcN can enhance the efficacy of PD-1 checkpoint blockade immunotherapy by amplifying tumor-specific humoral immunity.