Background: Adenocarcinoma of the urinary bladder (AUB), encompassing urachal adenocarcinoma (UAC) and primary bladder adenocarcinoma (BAC), poses a formidable clinical challenge with few therapeutic options. Immunotherapy holds emerging promise but is hindered by an insufficient understanding of the tumor immune microenvironment (TIME). The objective of the study was to delineate the immune composition and spatial architecture of the AUB tumor ecosystem and their implications for patient prognosis and immunotherapy. Methods: We compiled clinical data from 177 AUB cases (112 UAC, 65 BAC). A total of 104 samples underwent single or multiplex immunohistochemistry on tissue microarrays, enabling immunophenotyping and spatial analysis. Seven fresh surgical tumor tissues were subjected to single-cell RNA sequencing (scRNA-seq). Immunotherapy efficacy was assessed in 21 real-world AUB patients. Immune phenotypes and spatial metrics were derived and correlated with disease outcomes. Findings: UAC and BAC exhibited similar clinicopathological and prognostic profiles. Digital pathology-based immunotypes and immunoscores stratified AUB patients into different risk groups with distinct survival. Despite ubiquitous and abundant immune infiltration, scRNA-seq revealed an immunologically inert status with limited tumor-immune interactions. Spatial analysis identified eight cellular neighborhoods (CNs) across the AUB cohort, demonstrating predominantly immune-depleted tumor niches. Three spatial archetypes (SAs) were further defined and could predict patient prognosis. Real-world immunotherapy data confirmed suboptimal performance, with only 16.7% achieving pathological complete response in neoadjuvant settings and no full remission in metastatic cases. Interpretation: This multifaceted depiction of immune composition and spatial architecture elucidates the constrained efficacy of current immunotherapy in AUB. Our work presents the first spatial immunity-based risk stratification for AUB and may guide the development of rational immunotherapeutic strategies.

Neuroendocrine bladder carcinoma (NEBC) is a rare but highly aggressive histologic subtype of bladder cancer with poor prognosis, often driven by delayed diagnosis and limited therapeutic options; despite widespread use of next-generation sequencing, its cellular origin remains unclear and controversial. We aimed to synthesize up-to-date molecular and immune features of NEBC and translate them into practical guidance for diagnosis and treatment. We performed a narrative review of English-language studies indexed in PubMed and Web of Science (January 2000–August 2025) using predefined keywords, integrating genomic, transcriptomic, immunohistochemical, and clinical outcome data. Key findings indicate frequent co-occurrence and probable common clonal origin with urothelial bladder carcinoma, with hallmark TP53 and RB1 alterations, prevalent APOBEC-driven mutagenesis, and recurrent TERT promoter mutations; tumor mutation burden is heterogeneous but can be high. Despite this, NEBC commonly exhibits an immune-cold or immune-excluded microenvironment characterized by low PD-L1 expression and T-cell dysfunction, which may blunt responses to immune checkpoint inhibitor monotherapy. Diagnostic practice still relies on morphology supported by immunohistochemistry (synaptophysin, chromogranin A, CD56, GATA3), with emerging tools such as INSM1 and a decision-tree model using synaptophysin, CD117, and GATA3 that improve accuracy. Therapeutically, neoadjuvant chemotherapy—most commonly EP or IA—followed by radical cystectomy improves outcomes compared with initial cystectomy alone, while metastatic disease is typically managed with EP chemotherapy and radiotherapy with limited durability. Early data support immunotherapy, particularly immune checkpoint inhibitors, and suggest potential benefit from chemoimmunotherapy; a prospective trial of neoadjuvant anti-PD-L1 plus EP is underway, and antibody-drug conjugates and bladder-sparing multimodality strategies are emerging. In conclusion, comprehensive molecular and immune characterization is critical to refine diagnosis, optimize patient selection, and accelerate prospective trials that evaluate neoadjuvant chemotherapy, chemoimmunotherapy, and targeted approaches in NEBC.

PURPOSE:Although trimodal therapy is currently the standard organ-sparing approach for muscle-invasive bladder cancer (MIBC), its clinical benefit is limited, and noninvasive biomarkers to guide dynamic decision-making are lacking. Here, we present a proof-of-concept study evaluating disitamab vedotin (RC48, a HER2-targeted antibody-drug conjugate) combined with toripalimab (JS001, anti-PD-1) and radiotherapy for bladder preservation in localized HER2-positive MIBC. PATIENTS AND METHODS:In the first-stage of an open-label phase II clinical trial (ClinicalTrials.gov identifier: NCT05979740), six patients were enrolled and received disitamab vedotin, toripalimab, and radiotherapy. Adverse events were documented according to the National Cancer Institute Common Terminology Criteria for Adverse Events (CTCAE v5.0). Tumor response was evaluated every 12 weeks by radiographic imaging, cystoscopy with biopsies, and urine cytology. In parallel, we performed longitudinal liquid biopsy analyses of circulating tumor DNA (ctDNA) and urinary tumor DNA (utDNA) using PredicineCARE assay. RESULTS:The combination was overall tolerable, with no grade 4 treatment-related adverse events or deaths. Five patients (83.3 %) achieved a complete response and remained recurrence-free. Notably, utDNA testing showed high accuracy in monitoring therapeutic effectiveness and enabled early detection of tumor relapse, whereas ctDNA was largely undetectable across blood samples. CONCLUSIONS:These findings establish the feasibility, efficacy, and potential biomarker utility of a novel bladder-preserving regimen, setting the stage for a paradigm shift in MIBC management.

Introduction Myocardial infarction (MI) is a leading cause of death worldwide. Immune cells play a significant role in the MI development. This study aims to identify a marker related to neutrophil for the diagnosis and early progression of MI.Methods Key genes were screened using three machine learning algorithms to establish a diagnostic model. A gene associated with the early progression of MI was identified based on single cell RNA sequencing data. To further validate the predictive value of the gene, the mouse models of MI were constructed. Immunofluorescence (IF) analysis demonstrated the co-expression of the gene with neutrophils. Immunohistochemistry (IHC) was performed to validate the role of the gene in the progression of MI.Results Neutrophils were identified and verified as the key infiltrating immune cells (IICs) involved in the onset of MI. A diagnostic panel with superior performance was developed using five key genes related to neutrophils in MI (AUC = 0.887). Among the panel, IL1R2 was found to early phase of MI, which was further corroborated by IHC in mouse models of MI.Conclusions This study suggests that IL1R2, which is specific to neutrophils, can predict the diagnosis and early progression of MI, providing new insights into the clinical management of MI.

p53 expression and mutational landscape in NEBC.

NMF analysis of NEBC mutational signatures.

PURPOSE:Neuroendocrine bladder cancer (NEBC) poses a formidable clinical challenge and attracts keen interests to explore immunotherapy as a viable treatment option. However, a comprehensive immunogenomic landscape has yet to be thoroughly investigated. EXPERIMENTAL DESIGN:Leveraging a long-term cohort of natural NEBC cases, we employed a multimodal approach integrating genomic (n = 19), transcriptomic (n = 3), single-cell RNA sequencing (n = 1), and immunohistochemical analyses (n = 34) to meticulously characterize the immunogenicity and immunotypes of primary NEBC tumors. Clinical, pathological, medical imaging, and treatment information was retrospectively retrieved and analyzed. RESULTS:Our study unveiled that despite a considerable mutational burden, NEBC was typically immunologically inactive, as manifested by 'immune-excluded' or 'immune-desert' microenvironment. Interestingly, a subset of mixed NEBC with concurrent urothelial bladder cancer (UBC) histology displayed an 'immune-infiltrated' phenotype with prognostic relevance. When compared to UBC, NEBC lesions were distinguished by a denser cellular composition and augmented peritumoral extracellular matrix, which might collectively impede lymphatic infiltration. As a result, single-agent immune checkpoint inhibitors demonstrated limited efficacy against NEBC, while pharmacologic immunostimulation with combination chemotherapy conferred a more favorable response. CONCLUSIONS:These new insights derived from genomic profiling and immune phenotyping pave the way for rational immunotherapeutic interventions in NEBC patients, with the potential to ultimately reduce mortality from this otherwise fatal disease.

Gene expression and immune repertoire in NEBC.

Clear cell renal cell carcinoma (ccRCC), the most common type of renal cell carcinoma (RCC), is not sensitive to traditional radiotherapy and chemotherapy. The polyphenolic compound Gallic acid (GA) can be naturally found in a variety of fruits, vegetables and plants. Autophagy, an intracellular catabolic process, regulates the lysosomal degradation of organelles and portions in cytoplasm. It was reported that autophagy and GA could affect the development of several cancers. Therefore, the aim of the present study was to evaluate the effects of GA on ccRCC development and clarify the role of autophagy in this process. In the present study, the effects of GA on the proliferation, migration and invasion of ccRCC cells were investigated in vitro by Cell Counting Kit-8, colony formation, flow cytometry, wound healing and Transwell migration assays, respectively. Additionally, the effects of GA on ccRCC growth and metastasis were evaluated using hematoxylin-eosin and immunohistochemical staining in vivo. Moreover, it was sought to explore the underlying molecular mechanisms using transmission electron microscopy, western blotting and reverse transcription-quantitative PCR analyses. In the present study, it was revealed that GA had a more potent viability inhibitory effect on ccRCC cells (786-O and ACHN) than the effect on normal renal tubular epithelial cell (HK-2), which demonstrated that GA selectively inhibits the viability of cancer cells. Furthermore, it was identified that GA dose-dependently inhibited the proliferation, migration and invasion of ccRCC cells in vitro and in vivo. It was demonstrated that GA promoted the release of autophagy markers, which played a role in regulating the PI3K/Akt/Atg16L1 signaling pathway. All the aforementioned data provided evidence for the great potential of GA in the treatment of ccRCC.

Table S1. Quality control of sequencing data. Table S2. Representativeness of study participants. Table S3. Clinicopathological characteristics of 35 patients diagnosed with NEBC. Table S4. Summary of clinicopathological characteristics for the NEBC cohort. Table S5. Comparing clinical characteristics of patients in the NEBC and UBC cohort before and after PSM. Table S6. Whole genome sequencing statistics of 19 patients in NEBC cohort. Table S7. Density of immune cells in CT and IM region and immunotypes of patients in NEBC cohort. Table S8. Cell density and Ki-67 expresion in NEBC region and UBC region of patients diagnosed with mixed-UBC. Table S9. Clinical characteristics of metastatic NEBC patients receiving systemic treatment. Table S10. Clinical characteristics and density of immune cells of patients receiving neoadjuvant treatment.

Tumor purity, ploidy, and CNVs in NEBC.

BackgroundDue to the clinical characteristic of frequent recurrence in urothelial bladder cancer (UBC), patients face significant health impacts and economic burdens. Therefore, understanding the molecular mechanisms involved in UBC recurrence is crucial for reducing its recurrence rate. The aim of our study is to help urologists and clinical researchers gain a deeper understanding of the changes in the tumor microenvironment (TME) during UBC recurrence.MethodsWe collected 10 samples from primary and recurrent non-muscle-invasive bladder cancer (NMIBC) and performed single-cell RNA sequencing. By distinguishing and annotating cell subpopulations, we identified tissue preferences of some novel cell subgroups. Next, pseudotime trajectory analysis, cell-cell communication analysis, and function enrichment analysis were applied to evaluate the dynamic changes in the TME and biological functions. Finally, we validated the distribution of some of these cell subgroups using multiplex immunofluorescence experiments.ResultsWe identified a tumor-associated fibroblast (CAF) subtype with high COL18A1 expression that is highly expressed in recurrent NMIBC, suggesting that the stromal component of the tumor may play a crucial role in the recurrence process. Additionally, pseudotime trajectory analysis revealed a macrophage subtype with high IL-6 expression at the terminal stage of macrophage differentiation, exhibiting significant immunosuppressive features. This indicated the presence of immune exhaustion during NMIBC recurrence. Lastly, we found an upregulation of estrogen in recurrent urothelial cancer cells, which may partially explain the gender disparity observed in UBC.ConclusionThis study identified several cell subpopulations influencing NMIBC recurrence, which were heavily infiltrated in the TME of recurrent NMIBC. Additionally, the enrichment of estrogen in urothelial cancer cells from various sources suggested a role of sex hormones in NMIBC recurrence.

Background A substantial number of patients with bladder cancer fail to benefit from immune checkpoint inhibitors (ICIs). We aim to investigate whether the addition of other therapeutic modalities into immunotherapy may augment the immune reactivity, thereby improving the overall response rate. Methods We conducted a comprehensive assessment of the immunological changes following immunotherapy and chemotherapy, employing both single-cell RNA sequencing and bulk RNA sequencing analyses. Results The bladder cancer patient treated with ICIs exhibited a higher abundance of B cells and T follicular helper cells compared to the treatment-naïve patient. Analysis of public datasets and the in-house RJBLC-I2N003 cohort revealed the induction of tertiary lymphoid structure (TLS) neogenesis and maturation by immunotherapy. The IMvigor 210 study suggested that TLS could serve as a predictor of immunotherapy response and patient prognosis. In addition, genome-wide transcriptome data unveiled a shift towards the immune-enriched subtype over the desert subtype in patients receiving neoadjuvant chemotherapy. Notably, the proportions of CD20 + B cells, T follicular helper cells, and TLSs were significantly increased. In patients treated with a combination of neoadjuvant chemotherapy and ICIs, TLS positivity and maturity were improved compared to the baseline. Furthermore, neoadjuvant chemoimmunotherapy resulted in a higher rate of pathological complete response compared to monotherapies. Conclusions This work pinpointed the individual effect of immunotherapy and chemotherapy in fostering TLS development, and underscored the superior effectiveness of combined modalities in enhancing TLS maturation and response rates.

Radiographic evaluation and survival analysis in NEBC and UBC.

Histopathological characteristics of NEBC and survival analysis in NEBC and UBC.

Clear cell renal cell carcinoma (ccRCC), the most common type of renal cell carcinoma (RCC), is not sensitive to traditional radiotherapy and chemotherapy. The polyphenolic compound Gallic acid (GA) can be naturally found in a variety of fruits, vegetables and plants. Autophagy, an intracellular catabolic process, regulates the lysosomal degradation of organelles and portions in cytoplasm. It was reported that autophagy and GA could affect the development of several cancers. Therefore, the aim of the present study was to evaluate the effects of GA on ccRCC development and clarify the role of autophagy in this process. In the present study, the effects of GA on the proliferation, migration and invasion of ccRCC cells were investigated in vitro by Cell Counting Kit‑8, colony formation, flow cytometry, wound healing and Transwell migration assays, respectively. Additionally, the effects of GA on ccRCC growth and metastasis were evaluated using hematoxylin‑eosin and immunohistochemical staining in vivo. Moreover, it was sought to explore the underlying molecular mechanisms using transmission electron microscopy, western blotting and reverse transcription‑quantitative PCR analyses. In the present study, it was revealed that GA had a more potent viability inhibitory effect on ccRCC cells (786‑O and ACHN) than the effect on normal renal tubular epithelial cell (HK‑2), which demonstrated that GA selectively inhibits the viability of cancer cells. Furthermore, it was identified that GA dose‑dependently inhibited the proliferation, migration and invasion of ccRCC cells in vitro and in vivo. It was demonstrated that GA promoted the release of autophagy markers, which played a role in regulating the PI3K/Akt/Atg16L1 signaling pathway. All the aforementioned data provided evidence for the great potential of GA in the treatment of ccRCC.

目的 探索膀胱小细胞癌(SCCB)的临床特征、预后因素以及新辅助治疗疗效.方法 收集2012年1月-2022年9月上海交通大学医学院附属仁济医院泌尿科收治的47例 SCCB患者的临床资料.通过与尿路上皮癌肿瘤对比,探讨 SCCB临床病理特征;进一步通过非转移性膀胱小细胞癌队列(37例)分析其预后相关因素及新辅助治疗在非转移性 SCCB 治疗中的应用.结果 SCCB基线期肿瘤分期高,易发生肌层浸润(100％)及远处转移(21.3％),总生存期短(中位生存时间13.1个月,1年生存率53.7％,5年总生存率15.4％).肿瘤T分期为 SCCB患者的预后危险因素,术前接受新辅助治疗为独立保护因素,能显著降低其复发转移风险(HR=0.189,95％CI:0.051～0.697,P=0.012)及患者死亡风险(HR=0.177,95％CI:0.045～0.698,P=0.013),显著提高患者的无疾病生存期以及总生存时间.此外,相比单纯新辅助化疗,新辅助化疗联合免疫治疗能提高SCCB的病理完全缓解率.结论 SCCB恶性程度高、易转移、预后差,推荐将新辅助治疗联合膀胱根治性切除术作为SCCB的首选治疗方案.

Abstract Aims To systematically review the incidence and risk factors for recurrent FSGS after kidney transplantation. Methods We searched PubMed, Embase, Medline, Web of Science, the Cochrane Library, CNKI, CBMdisc, Wanfang, and Weipu for case-control studies related to recurrent FSGS from the establishment until October 2022. The protocol was registered on PROSPERO (CRD42022315448). Data were analyzed using Stata 12.0, with odds ratios (counting data) and standardized mean difference (continuous data) being considered as effect sizes. If the I 2 value was greater than 50%, the random-effects model was used; otherwise, a fixed-effects model was used. A meta-analysis on the incidence and risk factors for recurrent FSGS after kidney transplantation was performed. Results A total of 22 studies with 966 patients and 12 factors were included in the meta-analysis. There were 358 patients with recurrent FSGS and 608 patients without FSGS after kidney transplantation. The results showed that the recurrence rate of FSGS after kidney transplantation was 38% (95% CI: 31%–44%). Age at transplantation (SMD = −0.47, 95% CI −0.73 to −0.20, p = .001), age at onset (SMD = −0.31, 95% CI −0.54 to −0.08, p = .008), time from diagnosis to kidney failure (SMD = −0.24, 95% CI −0.43 to −0.04, p = .018), proteinuria before KT (SMD = 2.04, 95% CI 0.91 − 3.17, p < .001), related donor (OR 1.99, 95% CI 1.20 − 3.30, p = .007) and nephrectomy of native kidneys (OR 6.53, 95% CI 2.68 − 15.92, p < .001) were associated with recurrent FSGS, whereas HLA mismatches, duration of dialysis before KT, sex, living donor, tacrolimus use and previous transplantation were not associated with recurrent FSGS after kidney transplantation. Conclusions The recurrence of FSGS after kidney transplantation remains high. Clinical decision-making should warrant further consideration of these factors, including age, original disease progression, proteinuria, related donor, and nephrectomy of native kidneys.

e16605 Background: Bladder cancer is a common malignancy and detection of tumor recurrence is important to improve patient outcomes. Transurethral resection of bladder tumor (TURBT) is the standard treatment for early-stage bladder cancer, but the molecular detection of disease relapse remains a challenge. Reliable and non-invasive methods for monitoring the molecular changes in urine are crucial for timely intervention and disease management. Methods: In this study, 108 patients with early-stage bladder cancer who were selected for detection of disease relapse after the 1 st TURBT (median timing 4.8 months after TURBT) were prospectively enrolled. The molecular recurrence in these patients was detected from the urinary tumor DNA (utDNA) using a targeted next-generation sequencing (NGS) assay (PredicineCARE). This assay identifies somatic alterations in utDNA, including single nucleotide variations (SNVs), gene fusions and copy number variations (CNVs), and based on these variations, the tumor fraction was called to identify molecular relapse in the patients. Results: 77 out of 108 (71.3%) patients were found to be tumor positive in utDNA with a tumor fraction ≥ 0.5%. The assay identified 525 SNVs and 32 CNVs in the tumor positive patients. The most frequently mutated genes in the urine samples were TP53 (38%), TERT (32%), KMT2D (28%), ARID1A (19%), FGFR3 (19%), PIK3CA (14%) and ERBB2 (12%). Conclusions: This study demonstrates the feasibility of using urine ctDNA assay to detect molecular recurrence in early-stage bladder cancer patients treated by TURBT. This information can be used to improve patient outcomes by enabling early detection of tumor recurrence and guiding treatment decisions. The findings of this study highlight the promising potential of urine-based NGS assays for cancer detection and disease surveillance.