Abstract Background: Previous MCED studies have shown differences in detection performance across different aggressiveness cancer subtypes. However, the methylation profiles and ctDNA shedding patterns across different aggressiveness subtypes have not been analyzed. We systematically compared cell-free DNA (cfDNA) tumor fraction and methylation profiles across aggressiveness subtypes in three cancers to identify epigenetic biomarkers that differentiate tumor aggressiveness. Methods: Blood samples from a case-control study (NCT06217900) including 757 cancer cases were analyzed using a targeted methylation-based MCED test. We compared cfDNA tumor fraction and methylation profiles between aggressiveness subtypes, stage-matched small cell lung cancer (SCLC) (n=137) versus (vs) non-small cell lung cancer (NSCLC) (n=137), stage I invasive lung adenocarcinoma (IAC) (n=81) vs minimally invasive adenocarcinoma (MIA) (n=81), triple-negative breast cancer (TNBC) (n=151) vs non-TNBC (n=151), and intermediate/high-grade prostate cancer (Gleason Grade Group[GG] 2-5)(n=14) vs low-grade prostate cancer(GG1)(n=6). Tumor fraction was estimated using zero-inflated Negative Binomial model based on the distribution of methylation signals, and methylation profiles were assessed by calculating methylated/unmethylated (M/U) ratios between different aggressiveness subtypes. Marker comparisons were conducted using the Mann-Whitney U test with multiple testing corrections. Results: The sensitivity is higher in more aggressive subtypes (lung cancer 93.73% vs SCLC 99.27%; breast cancer 70.76% vs TNBC 81.46%; prostate cancer 40% vs intermediate and high-grad prostate cancer 42.86%). In lung cancer, SCLC exhibited a significantly higher cfDNA tumor fraction than NSCLC (Wilcoxon p = 2.24×10-24), with 41,136 markers (79.18% hypomethylated) showing elevated M/U ratios. In stage I lung adenocarcinoma, IAC demonstrated a higher cfDNA tumor fraction (Wilcoxon p = 2.55×10-6) and systematic methylation differences compared to MIA. In breast cancer, TNBC had a significantly higher cfDNA tumor fraction than non-TNBC (Wilcoxon p = 2.55×10-6), with 25,717 markers (80.32% hypomethylated) displaying increased M/U ratios. Among prostate cancers of the same stage, GG2-5 cases exhibited higher cfDNA tumor fraction (Wilcoxon p = 0.038) and systematic methylation alterations compared to GG1. Conclusion: Aggressive tumor subtypes consistently display elevated cfDNA tumor fraction and characteristic methylation profiles marked by increased M/U ratios. These findings suggest cfDNA methylation patterns are promising epigenetic biomarkers for distinguishing tumor aggressiveness, potentially improving cancer screening precision and reducing overdiagnosis. Citation Format: Kezhong Chen, Jian Huang, Dahong Zhang, Shu Wang, Hongxu Liu, Wenzhao Zhong, Xiangnan Li, Qiang Zhang, Zhigao Li, Jiaqi Liu, Ziqing Tian, Fei Zhou, Gongsheng Jin, Xudong Xiang, Zhigang Li, Hui Xie, Ya Wei, Guochun Zhang, Guolin Ye, Ming Cai, Junfeng Wang, Yan Zhang, Chao Cheng, Hefei Li, Desong Yang, Jianhong Lian, Sheng Huang, Tao Xu, Zengjun Wang, Xi Guo, Zhuowei Liu, Minfeng Chen, Yang Wang, Yue An, Yanzhan Yang, Min Li, Jing Liu, Baoliang Zhu, Yonghui Li, Xiaohui Wu, Fan Yang, Jun Wang. Epigenetic profiling identifies markers of aggressive cancer subtype using a targeted methylation-based multi-cancer early detection (MCED) blood test [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 1100.

4626 Background: SHR-1501, an IL-15 agonist fusion protein composing of a humanized antibody Fc region fused with IL-15 and IL-15Rα sushi domain, demonstrated promising efficacy, well tolerance, and acceptable safety in alone or in combination with BCG in patients with BCG-naive and BCG-unresponsive high-risk NMIBC ( ASCO 2025 ). Here, we report the updated results of this phase 1/2 study (NCT05410730). Methods: In the dose-escalation phase 1a and 1b parts, SHR-1501 monotherapy (200, 400, and 600 μg) or SHR-1501 (600 μg) in combination with BCG (120 mg) was administered to patients with high-risk NMIBC. In the phase 2 part, patients with BCG-naive NMIBC (cohort A), BCG-unresponsive NMIBC carcinoma in situ (CIS; cohort B), and BCG-unresponsive high-grade Ta/T1 NMIBC without CIS (cohort C) were enrolled to receive SHR-1501 (600 μg) plus BCG (120 mg). During the induction phase, all patients received weekly intravesical study treatment for 6 weeks. In the maintenance phase, instillations were administered weekly for three weeks at months 3, 6, 12, 18, and 24 following the initial induction dose. Primary endpoints were dose-limiting toxicity (DLT), maximum tolerated dose (MTD), and recommended phase 2 dose in phase 1a and 1b parts; and was complete response (CR) rate for cohort B and 12-mo disease-free survival (DFS) rate for cohorts A and C in phase 2 part. Results: As of Oct 31, 2025, 112 patients were enrolled (n=8 in phase 1a; n=6 in phase 1b; n=30, 25, and 43 in cohorts A, B, and C in phase 2). The median follow-up duration was 23.0 months (range 3.5-25.6) in patients with BCG-naive NMIBC, 6.5 months (range 2.6-21.2) in patients with BCG-unresponsive NMIBC CIS, and 13.5 months (range 2.5-23.1) in patients with BCG-unresponsive high-grade Ta/T1 NMIBC without CIS. In cohort B, the overall CR rate was 80.0% (20/25), the median DFS was 12.0 months (95% CI 6.0-NR). The 12-mo DFS rate was 90.3% (95% CI, 72.8-96.8) in patients with BCG-naive NMIBC and 62.7% (95% CI, 44.9-76.1) in patients with BCG-unresponsive high-grade Ta/T1 NMIBC without CIS. The 18-mo DFS rates were 90.3% (95% CI, 72.8-96.8) and 58.2% (95% CI, 39.6-72.9), respectively. Treatment-related adverse events (TRAEs) and grade 3 TRAEs occurred in 90 (86.5%) and 19 (18.3%) of 104 patients with SHR-1501 + BCG. The most common TRAEs were urinary tract infection (62.5%) and pollakiuria (35.6%). No TRAEs led to death. Conclusions: This updated analysis confirms the promising efficacy and manageable safety profile of SHR-1501 monotherapy or in combination with BCG in BCG-naive and BCG-unresponsive high-risk NMIBC patients. Two randomized, controlled phase 3 trials are underway in both BCG-unresponsive and BCG-naïve, high-risk NMIBC populations, with the recommended dose of 600 μg SHR-1501 plus BCG. Clinical trial information: NCT05410730 .

Sunitinib, a tyrosine kinase inhibitor, exerts its crucial therapeutic effect in renal cell carcinoma (RCC) primarily by inhibiting tumor angiogenesis and cellular proliferation. However, despite its initial efficacy, therapeutic resistance invariably develops in many RCC patients, underscoring the urgent need for novel strategies to overcome this limitation. We constructed patient-derived xenograft (PDX) models using surgically resected RCC tissues and subsequently established stable sunitinib-resistant and sunitinib-sensitive PDX models. Proteomic profiling was performed to identify key regulators involved in sunitinib resistance. Mechanistic studies were conducted to investigate the molecular interactions among DHODH, TRIM28, and VEGFA. The therapeutic potential of lisaftoclax was evaluated in combination with sunitinib. Proteomic profiling identified DHODH as a key regulator in the development of sunitinib resistance in RCC. Mechanistically, DHODH competes with the E3 ligase TRIM37 for binding to TRIM28, thereby stabilizing TRIM28 by inhibiting its ubiquitination. TRIM28 subsequently activates VEGFA transcription, which promotes sunitinib resistance in RCC. Lisaftoclax, a small-molecule inhibitor that disrupts the DHODH-TRIM28 interaction, potentiates sunitinib efficacy and exerts a synergistic therapeutic effect. Collectively, our findings identify DHODH as a critical therapeutic target for overcoming sunitinib resistance in RCC and provide a novel strategy for the treatment of RCC.

BACKGROUND:Clear cell renal cell carcinoma remains a therapeutic challenge due to tumor heterogeneity and limited biomarkers. Proliferating cell subpopulations in the tumor microenvironment contribute to cancer progression, yet their transcriptional features and clinical relevance in ccRCC are not fully characterized. METHODS:We integrated single-cell RNA-seq (GSE156632) and bulk transcriptomic data (TCGA-KIRC, EMTAB1980, CPTAC). Proliferating (G2/M-S phase) cells were identified through cell cycle analysis, and the Scissor algorithm was applied to select phenotype-associated malignant subpopulations. A prognostic model was constructed using 117 machine-learning algorithms and validated through multi-omics analysis. Functional assays including CCK-8, colony formation, wound healing, Transwell, and in vivo xenograft models were performed to validate key targets. The interaction between ISG15 and KPNA2 was investigated using co-immunoprecipitation, immunofluorescence, and ubiquitination assays. The role of ISG15 in regulating KPNA2 stability via the ubiquitin-proteasome pathway was examined using cycloheximide chase experiments and proteasome inhibition. RESULTS:We identified proliferative Scissor+ cells in ccRCC scRNA-seq data and derived 108 proliferative-associated genes. A prognostic model comprising 11 genes was constructed via Lasso+plsRcox, demonstrating robust predictive performance across multiple cohorts. High-risk patients exhibited enriched HIF-1/VEGF signaling, elevated TMB, immunosuppressive microenvironments, and increased TIDE scores. ISG15 knockdown suppressed proliferation, migration, and invasion in ccRCC cells by promoting KPNA2 ubiquitination and degradation, which was rescued by KPNA2 overexpression.

Intrinsic resistance to sunitinib in advanced renal cell carcinoma (RCC) remains a major barrier to improving patient survival outcomes. However, the molecular mechanisms driving this resistance remain incompletely elucidated. In this study, we first observed elevated glutamine levels in sunitinib-resistant RCC models; notably, glutamine deprivation substantially impaired the growth and proliferation of RCC cells. We further demonstrated that abnormal upregulation of GFPT2-a key enzyme in glutamine metabolism-was associated with reduced sunitinib sensitivity and enhanced drug resistance in RCC. Mechanistically, we uncovered that GFPT2 modulates cellular O-GlcNAcylation levels, which in turn enhances the stability and nuclear translocation of YAP1-ultimately contributing to reduced sunitinib sensitivity. In addition, we also identified an additional non-metabolic role of GFPT2: it directly interacts with the Kelch domain of KEAP1, thereby reducing NRF2 binding to this domain and suppressing NRF2 ubiquitination-dependent degradation. Consequently, this regulatory cascade dysregulates the transcription of downstream antioxidant genes (e.g., HMOX1 and NQO1), ultimately driving NRF2-dependent sunitinib resistance in RCC. Critically, this KEAP1-NRF2 axis-mediated mechanism operates independently of GFPT2's metabolic role in regulating O-GlcNAcylation. Collectively, our findings demonstrate that GFPT2 modulates sunitinib sensitivity and drives drug resistance in RCC via dual mechanisms: a metabolic pathway (O-GlcNAcylation-YAP1) and a non-metabolic pathway (KEAP1-NRF2). Targeting the non-metabolic functions of GFPT2 thus holds promise for enhancing sunitinib sensitivity in RCC while potentially mitigating treatment-related side effects.

Background Sunitinib resistance remains a major clinical challenge in renal cell carcinoma, and the underlying molecular mechanisms are incompletely understood. SP5, a member of the specificity protein transcription factor family, has been implicated in tumor progression, but its role in sunitinib resistance and angiogenesis in RCC is unclear. Methods We integrated bulk RNA-seq datasets (GSE172165, GSE173572, GSE216494) and single-cell transcriptomics (GSE156632) to identify sunitinib resistance-associated differentially expressed genes. A prognostic model was constructed using 117 machine-learning algorithms and validated across multiple independent cohorts. Functional assays, including CCK-8, colony formation, transwell, tube formation, CUT&RUN, and xenograft models, were performed. Virtual screening and molecular docking identified suramin as a potential SP5 inhibitor. Results We identified SP5 as a core resistance-related gene. The risk model effectively stratified patients, with high-risk individuals showing worse prognosis, elevated TMB, and immunosuppressive microenvironments. Mechanistically, SP5 directly bound to the VEGFA promoter to upregulate VEGFA transcription, activating AKT phosphorylation via autocrine and paracrine loops, thereby promoting angiogenesis and sunitinib resistance. SP5 knockdown suppressed proliferation, migration, invasion, and tube formation, and resensitized cells to sunitinib. Suramin disrupted the SP5–VEGFA interaction, synergized with sunitinib to inhibit tumor growth, and reduced VEGFA secretion. Conclusion Our findings establish SP5 as a critical driver of sunitinib resistance and identify suramin as a promising therapeutic strategy.

ObjectivesAbout 20% of renal masses are proved to be benign after surgery. We aimed to develop a strategy based on radiomics to differentially diagnose renal masses.Methods: In this retrospective study, 726 renal masses (54 benign and 672 malignant) were collected from two hospitals in China and two public databases. Two diagnostic models, kidney mass diagnosis model (KMDM) and kidney cancer subtypes diagnosis model (KCSDM), were developed for differential diagnosis of renal cell carcinoma and their subtypes. Each model was divided into three units (radiomics model, clinical model, and integrated model) and evaluated in the training set and two independent validation sets.MethodsIn this retrospective study, 726 renal masses (54 benign and 672 malignant) were collected from two hospitals in China and two public databases. Two diagnostic models, kidney mass diagnosis model (KMDM) and kidney cancer subtypes diagnosis model (KCSDM), were developed for differential diagnosis of renal cell carcinoma and their subtypes. Each model was divided into three units (radiomics model, clinical model, and integrated model) and evaluated in the training set and two independent validation sets.ResultsA total of 39 radiomic variables were used for the KMDM, and 78 variables for the KCSDM. The radiomics model performed better than the clinical model in both KMDM and KCSDM. The integrated model of KMDM showed the best performance (AUC of 0.937 in training set, 0.942 in both two validation sets), when compared to the other two models and two experts. A cutoff value of risk of malignant index (RMI=1.661) was designated to distinguish the benign and malignant renal masses with a sensitivity of 0.929 and a specificity of 0.833 in validation sets. Similar results were found in the integrated model of KCSDM.ConclusionThe integrated model can noninvasively distinguish benign from malignant renal masses and renal cell carcinoma subtypes with satisfactory sensitivity and specificity.

BackgroundThere is no systematic classification of renal vascular injuries conducted for severe post-percutaneous nephrolithotomy (PCNL) bleeding.AimThe aim of the present study was to explore the various types of artery injury and clinical characteristics of patients who underwent transcatheter angioembolization (TAE) after PCNL.MethodsA retrospective analysis was performed on 52 patients who underwent renal arteriography (RA) because of severe bleeding after PCNL between April 2009 and December 2023. Among the patients, 38 underwent TAE due to positive RA results. Clinical data on the TAE patients, such as gender, age, body mass index, TAE interval, hemoglobin (Hb) decrease, operation time, stone size, the number and size of tracts, and clinical bleeding type, were summarized. The types of artery injury in TAE patients and their relationships with clinical characteristics were analyzed.ResultsRetrospective analysis revealed that, among the 38 TAE patients (32 males and 6 females), the mean TAE interval, average Hb decrease, mean tract number, and mean tract size reached 5.00 (6.25) days, 44.50 (24.50) g/L, 1 (0.25), and F20(6), respectively. Among the TAE patients, four kinds of vascular injury were observed, namely, 18 cases of pseudoaneurysm (PA), 12 cases of arteriocaliceal fistula (ACF), 7 cases of arteriovenous fistula (AVF), and 1 case of arterioperirenal fistula (APF). Analysis of the clinical characteristics of the three types of vascular injury (PA, ACF, and AVF) revealed that the number of tracts was the only factor that differed.ConclusionThe RA results indicate that the types of postoperative renal artery injury mainly include PA, ACF, AVF, and APF, and the number of tracts may be related to the type of vascular injury.

BACKGROUND:The underexplored potential of PD-L1 blockade in advanced renal cell carcinoma highlights an urgent need for novel agents. This trial aimed to compare benmelstobart (a novel PD-L1 inhibitor) plus anlotinib with sunitinib as first-line treatment for advanced renal cell carcinoma. METHODS:ETER100 was a multicentre, randomised, open-label, phase 3 trial conducted at 37 medical sites in China. We included patients aged 18-80 years, who had previously untreated, advanced, clear-cell renal cell carcinoma, and an Eastern Cooperative Oncology Group performance status of 0 or 1. We randomly assigned (1:1) patients to receive either benmelstobart (intravenous, 1200 mg, once every 3 weeks) plus anlotinib (oral, 12 mg, once daily for the first 2 weeks of a 3-week cycle) or sunitinib (oral, 50 mg, once daily for the first 4 weeks of a 6-week cycle) until disease progression, unacceptable toxicity, investigator's decision, or patient withdrawal. Randomisation was done centrally with stratified block randomisation (block size 4) and stratified by International Metastatic Renal Cell Carcinoma Database Consortium risk. The primary endpoint was progression-free survival as assessed by blinded independent central review according to the Response Evaluation Criteria in Solid Tumours version 1.1 in the full analysis set (ie, randomly assigned patients who received at least one dose of study drug without the violation of key inclusion criteria) and per-protocol set (ie, randomly assigned patients who received at least one cycle of protocol treatment without major protocol violations and had at least one efficacy assessment). In this Article, we report the results of a prespecified interim analysis. This ongoing study, closed to recruitment, is registered with ClinicalTrials.gov, NCT04523272. FINDINGS:Between Aug 25, 2020, and Feb 6, 2023, we assessed 687 patients for eligibility, 531 (77%) of whom were randomly assigned to receive either benmelstobart plus anlotinib (266 [50%] patients) or sunitinib (265 [50%] patients). 527 (99%) patients were included in the full analysis set (263 [50%] patients who received benmelstobart plus anlotinib and 264 [50%] who received sunitinib). All patients were Chinese (400 [76%] men and 127 [24%] women), with a median age of 60 years (IQR 54-67). As of the cutoff date (Jan 31, 2024), the median follow-up was 22·8 months (IQR 15·2-29·7). In the full analysis set, median progression-free survival was significantly longer with benmelstobart plus anlotinib than with sunitinib (19·0 months [95% CI 15·3-22·8] vs 9·8 months [8·4-12·4]; hazard ratio [HR] 0·53 [95% CI 0·42-0·67]; p<0·0001). In the per-protocol set, median progression-free survival was 19·0 months (16·5-22·8) in the benmelstobart-anlotinib group versus 11·0 months (8·5-13·6) in the sunitinib group (HR 0·55 [0·43-0·70]; p<0·0001). The most common grade 3 or worse treatment-related adverse event was hypertension (occurring in 91 [34%] of 264 patients in the benmelstobart-anlotinib group vs 55 [21%] of 264 in the sunitinib group). Serious treatment-related adverse events occurred in 63 (24%) patients in the benmelstobart-anlotinib group and in 42 (16%) patients in the sunitinib group. In the benmelstobart-anlotinib group, three (1%) deaths occurred due to treatment-related adverse events (one each with cardiac-respiratory arrest, unknown reason, and renal failure) and no deaths occurred in the sunitinib group. INTERPRETATION:Benmelstobart plus anlotinib improved progression-free survival compared with sunitinib among patients with previously untreated, advanced clear-cell renal cell carcinoma. These findings suggest the potential of benmelstobart plus anlotinib as a treatment option for this population. FUNDING:Chia Tai Tianqing Pharmaceutical Group and CSCO Clinical Oncology Research Foundation. TRANSLATION:For the Chinese translation of the abstract see Supplementary Materials section.

Objective: Seminal vesiculitis is a prevalent infectious disease in the genitourinary system, but its intrinsic pathogenesis and molecular features remain unclear. Methods: In this study, we utilized a next-generation sequencing technology to demonstrate transcriptomic landscapes of clinical specimens from seminal vesiculitis patients. We identified enriched Go Ontology terms, pathway maps, processes, toxicity, and metabolic networks of seminal vesiculitis. Results: Glutathione S-transferase activity is highly enriched, indicating its essential role in the disease's development. Furthermore, we established inflammation models of the seminal vesicle and verified the crucial function of glutathione S-transferase activity in these models. Conclusion: Glutathione S-transferase serves an essential role in seminal vesiculitis patients. Our results shall provide a robust platform for further intensive studies of seminal vesiculitis.

4536 Background: Dual immune checkpoint inhibitors (ICIs) or ICIs plus VEGF-directed therapies, have been approved as first-line treatment in patients (pts) with advanced renal cell carcinoma (RCC). The phase 3 ETER100 trial showed that benmelstobart (PD-L1 blockade) plus anlotinib improved the progression-free survival (PFS) (19.0 months 9.8 months) and objective response rate (ORR) (71.6% vs 25.1%) of advanced clear cell RCC (ccRCC) pts significantly. Pts with factors, such as intermediate-poor International Metastatic Renal Cell Carcinoma Database Consortium (IMDC) risk, liver metastasis, or bone metastasis were considered to have a poor prognosis. Here we report PFS and ORR in clinically relevant subgroups. Methods: ETER100 (NCT04523272) was a multicentre, randomised, open-label, controlled phase 3 trial conducted at 37 sites in China. Eligible patients were randomly assigned in a 1:1 ratio using stratified block randomisation to receive benmelstobart plus anlotinib or sunitinib. Randomisation was stratified according to the International Metastatic Renal Cell Carcinoma Database Consortium (IMDC) risk (favourable [score of 0], intermediate [score of 1-2], or poor risk [score of 3-6]). PFS analyses of clinically relevant subgroups were assessed using Kaplan-Meier method and the 95% CIs of response rate were calculated with the Clopper-Pearson method. Results: Overall, 527 pts received the trial treatments (264 in the benmelstobart-anlotinib group and 263 in the sunitinib group) and were evaluated for efficacy. A total of 454 (86%) pts had intermediate-poor IMDC risk, 62 (12%) pts had liver metastasis and 111(21%) had bone metastasis. Data cutoff for the interim analysis occurred on January 31, 2024. The median follow-up was 22.8 months. Benmelstobart plus anlotinib significantly improved PFS of subgroups with intermediate-poor IMDC risk (17.0 months [95% CI 14.0-20.1] vs 9.7 months [8.0-11.3], HR 0.55, 95% CI 0.43-0.72; p < 0.0001), liver metastasis (11.9 months [95% CI 5.8-NE] vs 5.4 months [1.5-6.7], HR 0.44, 95% CI 0.23-0.85; p < 0.0121), or bone metastasis (19.5 months [95% CI 16.5-27.2] vs 8.3 months [4.2-19.8], HR 0.52, 95% CI 0.30-0.89; p < 0.0154). ORR of benmelstobart-anlotinib group was significantly higher in the subgroups with intermediate-poor IMDC risk (70.0% [95%CI, 63.6- 75.9] vs 21.6% [16.4-27.5]), liver metastasis (60.0% [95%CI, 42.1-76.1] vs 7.4% [0.9-24.3]) and bone metastasis (63.2% [95%CI, 49.3-75.6] vs 16.7% [7.9-29.3]). Conclusions: The RCC pts with a poor prognosis such as intermediate-poor IMDC risk, liver metastasis and bone metastasis could significantly benefit from benmelstobart plus anlotinib. Clinical trial information: NCT04523272 .

Sunitinib resistance is one of the main reasons for the poor prognosis of clear renal cell carcinoma (ccRCC). Moreover, Stress granules (SGs) was found to enhance the stress adaptation capability of tumor cells, becoming an important mechanism for drug resistance in various cancers. We developed sunitinib-resistant patient-derived xenograft (PDX) and organoid (PDO) models to investigate sunitinib resistance in ccRCC. Proteomic analysis identified UBAP2L as a key mediator of this resistance. To explore its role in stress granule formation and sunitinib resistance, we conducted both in vitro and in vivo studies. We further elucidated the regulatory mechanisms of UBAP2L O-GlcNAcylation using immunoprecipitation, mass spectrometry, modification-based proteomics, RNA sequencing (RNA-seq), and RNA immunoprecipitation sequencing (RIP-seq). In this study, enrichment of UBAP2L was elucidated to be significantly associated with sunitinib-resistant ccRCC patient-derived xenograft (PDX) model. Functional experiments showed that UBAP2L protected ccRCC from apoptosis and promoted ccRCC prolifecation and angiogenesis upon sunitinib treatment, thus enhancing drug resistance of ccRCC cells. Furthermore, mechanistic investigation demonstrated that O-GlcNAcylation of UBAP2L promoted its protein stability via inhibiting TRIM37-mediated ubiquitination and it regulated stress granule formation, thereby enhancing the mRNA stability of Melk and activating the PI3K signaling pathways. These results validated the significant roles of O-GlcNAcylation of UBAP2L in ccRCC sunitinib resistance, which provided an innovative theoretical basis for the clinical diagnosis and therapy of ccRCC.

BACKGROUND:Clear cell renal cell carcinoma represents the most prevalent histological subtype of renal malignancy Emerging evidence underscores the critical involvement of mitochondrial dysfunction in oncogenesis and tumor progression. In this study, we systematically identified and characterized mitochondrial-associated proteins participating in fundamental biological processes, with the objective of elucidating their mechanistic contributions to ccRCC pathogenesis. METHOD:Through integrative analysis of multi-omics data from TCGA-KIRC and GEO datasets, we systematically evaluated the prognostic value of mitochondrial-associated differentially expressed genes derived from the MitoCarta3.0 database. Employing an ensemble machine learning approach encompassing 10 base algorithms and their combinatorial variants, we developed and optimized a robust prognostic risk model. Subsequent experimental validation focused on the core model gene FKBP10, investigating its functional role in ccRCC pathogenesis through comprehensive in vivo analyses. RESULT:We identified 102 mitochondrial-related differentially expressed genes showing significant tumor-normal tissue expression differences and strong prognostic associations. The LASSO+GBM algorithm was selected as the optimal model, from which we derived 7 core prognostic genes. Comprehensive characterization of high- and low-risk groups revealed distinct patterns in pathway activation, mutational profiles, immune microenvironment composition, and therapeutic vulnerabilities. Experimental validation confirmed that the key gene FKBP10 promotes malignant phenotypes in tumor cells through IL-6/JAK/STAT3 pathway activation.

MicroRNAs have been implicated in regulating diverse cellular pathways. Emerging evidence indicates that miR-143 plays causal roles in cancer tumorigenesis as a tumor suppress gene; however, its role in prostate cancer tumorigenesis remains largely unknown. The aims of this study were to verify the effect of miR-143 on proliferation and migration abilities of prostate cancer cells. The expression level of miR-143 and its target gene KRAS were measured by realtime PCR and western blotting, respectively. Effects of miR-143 in cell proliferation, migration and chemosensitivity were evaluated by MTT assay, FACS cell cycle analysis, colony formation assay, and transwell migratory assay. Our results revealed an inverse correlation of expression between miR-143 and KRAS protein in prostate cancer samples (Pearson’s correlation scatter plots: R = −0.707, P < 0.05). Moreover, over-expression of miR-143 in prostate cancer cells suppressed their proliferation and migration and increased their sensitivity to docetaxel by targeting EGFR/RAS/MAPK pathway. These findings suggest that miR-143 plays an important role in prostate cancer proliferation, migration and chemosensitivity by suppressing KRAS and subsequent inactivation of MAPK pathway, which provides a potential development of a new approach for the treatment of prostate cancer.

4606 Background: BCG is the standard therapy after transurethral resection of bladder tumor for high-risk NMIBC. IL-15 agonists can enhance the immune response induced by BCG via stimulating the proliferation and activation of natural killer cells and CD8+ cytotoxic T cells, without inducing regulatory T cells. SHR-1501 is an IL-15 agonist fusion protein, composed of a humanized antibody Fc region fused with IL-15 and IL-15Rα sushi domain. In this phase 1/2 study, we assessed the safety, tolerability, and efficacy of SHR-1501 in patients (pts) with high-risk NMIBC. Methods: The study comprised dose-escalation phase 1a and 1b parts of SHR-1501 alone or in combination with BCG in pts with high-risk NMIBC, followed by a phase 2 part of SHR-1501 plus BCG in multiple cohorts, including pts with BCG-naive NMIBC (cohort A), BCG-unresponsive NMIBC carcinoma in situ (CIS; cohort B), and BCG-unresponsive high-grade Ta/T1 NMIBC without CIS (cohort C). All pts received intravesical study treatment weekly for 6 weeks during induction period. During maintenance period, instillations occurred weekly for the first 3 weeks at 3, 6, 12, 18, and 24 months after the initial induction instillation. Primary endpoints were dose-limiting toxicity (DLT), maximum tolerated dose (MTD), and recommended phase 2 dose in phase 1a and 1b parts; and was complete response (CR) rate for cohort B and 12-mo disease-free survival (DFS) rate for cohorts A and C in phase 2 part. Results: As of Sep 7, 2024, 84 pts were enrolled (n = 8 in phase 1a; n = 6 in phase 1b; n = 29, 17, and 24 in cohorts A, B, and C in phase 2). In phase 1a part of SHR-1501 alone (200, 400, and 600 μg) and phase 1b part of SHR-1501 (600 μg) plus BCG (120 mg), no DLTs were observed, and MTD was not reached. Thus, 600 μg of SHR-1501 plus 120 mg of BCG was used in phase 2 part. Treatment-related adverse events (TRAEs) occurred in 4 (50.0%) of 8 pts with SHR-1501 and 53 (69.7%) of 76 pts with SHR-1501 + BCG. Grade 3 TRAEs were reported in 1 (12.5%) pt with SHR-1501 (urinary tract infection) and 7 (9.2%) pts with SHR-1501 + BCG (urinary tract infection and hypertension occurred in > 1 pt). No grade 4 or 5 TRAEs were reported. No serious TRAEs occurred. Of the efficacy evaluable pts in cohort B, the CR rate at 3 or 6 months was 90.9% (10/11). In cohorts A and C, the 12-month DFS rate was not reached. The 9-month DFS rate was 94.4% (95% CI, 66.6-99.2) in cohort A and 53.9% (95% CI, 15.5-81.4) in cohort C. Conclusions: SHR-1501 alone or in combination with BCG was well-tolerable and demonstrated a favorable efficacy in BCG-naive and BCG-unresponsive high-risk NMIBC pts, supporting further investigations. Clinical trial information: NCT05410730 .

Background: Clear cell renal cell carcinoma (ccRCC) is a prevalent and aggressive subtype of kidney cancer, often associated with metastasis and recurrence. Identifying key genes involved in ccRCC progression is critical for improving treatment strategies and patient outcomes. Methods: We performed a large-scale genome-wide CRISPR screening to identify genes crucial to ccRCC progression using the DepMap database. For discovery and validation, we integrated multi-omics data from The Cancer Genome Atlas (TCGA), GEO, and the NJMU-ccRCC clinical cohort. Bioinformatics analyses, including differential expression, pathway enrichment, and protein-protein interaction network analysis, were conducted to elucidate the biological functions. To validate our findings, we employed immunohistochemistry, qRT-PCR, and various cellular assays to investigate the role of PRC1 in ccRCC. Results: CRISPR screening identified PRC1 as a key gene significantly overexpressed in ccRCC tissues from the DepMap database. Elevated PRC1 expression was associated with poor overall survival, disease-specific survival, and progression-free interval. Silencing PRC1 in ccRCC cell lines inhibited cell proliferation, migration, and colony formation. Functional enrichment analyses revealed that PRC1 is involved in essential processes such as cell cycle regulation, mitosis, and cytokinesis. Additionally, PRC1 expression was correlated with the activation of the Wnt/β-catenin pathway, suggesting that PRC1 plays a pivotal role in tumor progression. Conclusion: PRC1 emerges as a promising biomarker and therapeutic target for ccRCC. Elevated PRC1 expression is associated with poor prognosis, and its inhibition suppresses ccRCC cell proliferation and migration. Our findings underscore the crucial role of PRC1 in ccRCC progression and highlight the need for further investigation into its molecular mechanisms and therapeutic potential.

INTRODUCTION:Polycyclic aromatic hydrocarbons (PAHs) are a group of chemicals that can induce oxidative stress and related cytotoxicity. Whether urinary concentrations of PAHs have effects on overactive bladder (OAB) in the general population is still unclear. This study investigated the associations between urinary PAHs and OAB. METHODS:7,146 adults aged over 20 who participated in the US National Health and Nutrition Examination Survey 2005-2016 were studied. The impact of the six PAHs on OAB was evaluated by multivariate logistic regression, and percent changes related to different quartiles of those six PAH levels were calculated. Confounders including age, logarithmic urinary creatinine, gender, race, body mass index, educational level, marriage, poverty income ratio, diabetes, hypertension, and metabolic syndrome were controlled. RESULTS:There is a significant positive correlation between urinary concentrations of the six PAHs we include in the study and the occurrence of OAB. Furthermore, individuals with higher PAH levels also reported a more severe OAB symptom score (OABSS). CONCLUSIONS:Our findings revealed that adult men in the USA with higher urinary PAHs had a higher risk of OAB incidence. These findings suggest the importance of strong environmental regulation of PAHs to protect population health. However, the underlying mechanisms still need further exploration.

Over the years, numerous epidemiological studies have shown that chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) promotes erectile dysfunction. Nonetheless, the precise underlying mechanism remains to be fully clarified. The objective of this research was to identify crucial signaling pathways responsible for CP/CPPS-induced erectile dysfunction. Thirty 8-week-old male Sprague-Dawley rats were randomly assigned to either the CP/CPPS model group or the control group. The CP/CPPS rat model was established through subcutaneous injection of a combination of rat prostate protein and Freund's adjuvant. Penile erectile function assessment was conducted 45 days after immunization through electrical stimulation of the cavernous nerve. RNA sequencing of the corpus cavernosum of the penis was then performed using the Kyoto Encyclopedia of Genes and Genomes and protein-protein interaction network analysis. Western blotting was performed on the cavernous tissue. Cell apoptosis assays, cell counting kit-8 assays, cell cloning assays, and Western blotting were conducted on rat endothelial cells. Erectile function was significantly lower in the CP/CPPS model group than in the control group (p < 0.001). Kyoto Encyclopedia of Genes and Genomes pathway analysis revealed that differentially expressed genes were predominantly enriched in the apoptosis pathway. Moreover, an increase in apoptosis in the rat corpus cavernosum, along with a decrease in the protein expression of CD31 (p = 0.0089) and eNOS (p = 0.0069) following CP/CPPS induction, was observed. In a protein-protein interaction network, Pitx2 was recognized as a central gene. The role of Pitx2 in regulating apoptosis was demonstrated in experiments using rat endothelial cell lines, and it was found to be regulated by the Wnt/beta-catenin pathway. This study highlights the occurrence of cavernous endothelial cell apoptosis in CP/CPPS-induced erectile dysfunction, and the potential mechanism of apoptosis may involve inhibition of the Wnt/beta-catenin/Pitx2 pathway.

Prostate cancer is a major public health challenge for men worldwide, being the second most common cancer diagnosis and the fifth leading cause of cancer-related deaths among men. The etiology of prostate cancer is multifactorial, with age, genetic predispositions, and lifestyle factors playing critical roles. The role of the peroxisome proliferator-activated receptors (PPARs) in prostate cancer remains complex and not fully elucidated. Transcriptomic data from The Cancer Genome Atlas (TCGA) were utilized for this study. The data were analyzed using single-sample Gene Set Enrichment Analysis (ssGSEA), Gene Ontology (GO) enrichment analysis, KEGG pathway analysis, and Gene Set Enrichment Analysis (GSEA). A prognosis-prediction model was constructed using Cox regression and LASSO analysis. Functional experiments, including Western blotting, quantitative PCR (qPCR), Cell Counting Kit-8 (CCK-8) assays, and EdU incorporation assays, were performed to validate the role of BUD23 in prostate cancer. Significant differences were observed in the immune microenvironment and HLA gene expression profiles between high and low PPAR expression groups in prostate cancer. The high PPAR group exhibited a less active immune microenvironment with higher fractions of immunosuppressive T regulatory cells (Tregs). A robust prognostic model identified key genes, including BUD23, associated with patient survival. Elevated BUD23 expression was correlated with more aggressive clinical features such as advanced pathological stages, nodal metastasis, and higher Gleason scores. Knockdown of BUD23 in PC-3 and LNCaP prostate cancer cell lines significantly inhibited cell proliferation. Western blotting and qPCR confirmed effective BUD23 knockdown, and CCK-8 and EdU assays demonstrated reduced cell proliferation. BUD23 knockdown resulted in significant reductions in PPAR-α, PPAR-β, and PPAR-γ protein levels, suggesting a regulatory axis between BUD23 and PPARs in prostate cancer. The study highlights the distinct roles of PPARα, PPARβ/δ, and PPARγ in prostate cancer progression and their potential as therapeutic targets. Elevated BUD23 expression is associated with aggressive prostate cancer features and patient survival, making it a potential prognostic biomarker. The knockdown of BUD23 not only inhibited cell proliferation but also reduced the expression of PPAR-related proteins, indicating a potential regulatory axis between BUD23 and PPARs. These findings suggest that targeting BUD23 could modulate PPAR signaling and inhibit tumor growth in prostate cancer.