OBJECTIVE:Acrylamide (AA), a widespread environmental and food-borne pollutant, has been classified as a probable human carcinogen. However, its specific role and underlying mechanisms in the progression of prostate cancer (PCa) remain poorly elucidated. This study aims to comprehensively investigate the effect of AA on PCa progression and its molecular mechanisms. METHODS:Differentially expressed AA-related genes (DEARGs) were identified by intersecting data from the Comparative Toxicogenomics Database (CTD) and The Cancer Genome Atlas (TCGA) database. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment analyses were performed to reveal biological functions of DEARGs. A prognostic signature was constructed using Lasso regression analysis. The expression and cellular localization of the key genes were analyzed using single-cell RNA sequencing (scRNA-seq) data. Additionally, the effects of glycidamide (GA) on PCa cells malignant phenotypes and tumor growth in vivo were experimentally validated. RESULTS:We identified a total of 165 DEARGs significantly associated with PCa. Functional analysis revealed they mainly enriched in PI3K-Akt and cAMP signaling pathway. The prognostic model constructed from the six DEARGs (CYP11A1, TUBB3, ALB, ESPL1, FOXN4, ISYNA1) demonstrated satisfactory predictive accuracy for biochemical recurrence-free survival (BCRFS) in PCa patients. scRNA-seq analysis delineated the specific expression of six target genes within the tumor microenvironment. It was found that expression of CYP11A1 was downregulated, while expressions of TUBB3, ALB, ESPL1, FOXN4 and ISYNA1 were upregulated after treatment with GA. In vitro and in vivo experiments confirmed that GA significantly enhanced proliferation, migration, invasion of PCa cells and tumor growth. RNA‑sequencing and functional rescue assays further revealed that the MAPK pathway may be a key mechanism by which GA promoted proliferation, invasion, and metastasis of PCa cells. CONCLUSION:This study successfully identified and validated an AA-related prognostic model to effectively predict BCRFS of PCa. The oncogenic effects of GA are primarily mediated by the activation of specific MAPK signaling pathways, elucidating a key mechanistic basis for its toxicity in PCa.

Background Varicocoele is a major cause of male infertility, yet its underlying molecular mechanisms and determinants of surgical efficacy remain unclear.Objective This study aimed to identify key proteins involved in varicocoele-related infertility and to investigate the expression and functional role of ubiquitin-specific peptidase 2 in spermatozoa and spermatogenic cells.Methods Ubiquitin-specific peptidase 2 expression in spermatozoa from varicocoele patients was analyzed before and after varicocelectomy by Western blotting. In vitro, ubiquitin-specific peptidase 2 was overexpressed or silenced in mouse GC-2 spermatocyte cells to assess mitochondrial autophagy (PINK1/Parkin pathway), autophagic flux, apoptosis, and reactive oxygen species. In vivo, a left-sided varicocoele rat model was established, and ubiquitin-specific peptidase 2 activity was inhibited by intratesticular injection of ML364. Testicular histopathology and sperm motility were evaluated.Results Ubiquitin-specific peptidase 2 protein expression was significantly reduced in spermatozoa after varicocelectomy. Ubiquitin-specific peptidase 2 overexpression enhanced PINK1/Parkin-mediated mitochondrial autophagy and protected spermatogenic cells from apoptosis, whereas ubiquitin-specific peptidase 2 knockdown impaired mitochondrial autophagy and increased cell apoptosis. In varicocoele rats, ubiquitin-specific peptidase 2 inhibition aggravated seminiferous tubule damage, reduced spermatogenic cell density, and impaired sperm motility. In human spermatozoa, ML364 treatment significantly decreased progressive motility without affecting sperm concentration.Conclusion Ubiquitin-specific peptidase 2 is essential for maintaining mitochondrial quality control and spermatogenic cell survival in varicocoele by regulating PINK1/Parkin-mediated autophagy and oxidative stress, highlighting its potential as a biomarker and therapeutic target for varicocoele-related male infertility.

Environmental polyethylene terephthalate micro- and nanoplastics (PET-MNPs) are biologically accessible contaminants, but their effects on clear cell renal cell carcinoma (ccRCC) progression remain unclear. Using 786-O and CAKI-1 cells and subcutaneous and orthotopic xenograft models, we found that PET-MNP exposure enhanced malignant phenotypes and tumor growth, accompanied by increased intracellular and mitochondrial ROS. Mitochondrial superoxide accumulation was associated with membrane depolarization, reduced ATP levels, ultrastructural abnormalities, and decreased SOD2 expression and Mn-SOD activity. Mechanistically, PET-MNP exposure increased AKT and inhibitory GSK3β Ser9 phosphorylation, reduced degradation-associated phosphorylation and ubiquitination of β-catenin, and enhanced β-catenin stability, nuclear accumulation, and transcriptional activity. Time-course analysis indicated that AKT activation preceded prominent GSK3β and β-catenin changes. MitoTEMPO, pharmacological AKT or β-catenin inhibition, AKT knockdown, β-catenin-S33Y rescue, and GSK3β-S9A experiments further supported the functional involvement of mitochondrial ROS-associated AKT/GSK3β/β-catenin regulation. ChIP-qPCR and RT-qPCR showed increased β-catenin recruitment to MYC and CCND1 regulatory regions and enhanced transcription. These findings support a role for mitochondrial ROS-associated AKT/GSK3β/β-catenin regulation in PET-MNP-promoted ccRCC progression.

PURPOSE:The purpose of this study is to construct and validate a neuroendocrine differentiation-related molecular model for predicting prognosis in patients with prostate cancer (PCa). MATERIALS AND METHODS:Transcriptome data for PCa were collected from the Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) websites. Differentially expressed neuroendocrine differentiation related genes (NDGs) were identified. By utilizing multivariate Cox analysis, a neuroendocrine differentiation-related molecular model for predicting prognosis was constructed and validated. The study investigated the novel model's association with the tumor immune microenvironment, clinicopathological characteristics, tumor stemness, and anticancer treatment sensitivity. Additionally, preliminary experimental verifications of Diencephalon / Mesencephalon Homeobox 1 (DMBX1) were conducted. RESULTS:Finally, we identified a total of 19 differentially expressed NDGs. A neuroendocrine differentiation-related molecular model was established and successfully validated both internally and externally. The high-risk group exhibited significantly poorer biochemical recurrence-free survival (BCRFS) in the training, testing, and validating cohorts. The areas under the receiver operating characteristic curves for the training, testing, and validating cohorts were 0.825, 0.719, and 0.729, respectively. The tumor immune microenvironment, clinicopathological features, tumor stemness, and anti-cancer drug sensitivity was significantly different between high and low-risk patients. Preliminary experiments revealed that higher expression of DMBX1 significantly enhanced the proliferation, migration, and neuroendocrine differentiation of PCa cells. CONCLUSION:This research developed a unique neuroendocrine differentiation-related molecular model that is highly suitable for predicting BCRFS. High DMBX1 expression may promote the development and neuroendocrine differentiation of prostate cancer.

Liquid-biopsy nucleic acid biomarkers hold strong promise for non-invasive cancer diagnostics, yet their ultralow abundance presents substantial analytical challenges. Conventional electrochemical biosensors rely on complex probe immobilization, whereas emerging homogeneous strategies remain susceptible to matrix interference and require large reaction volumes. Here, we report a probe immobilization-free, reagent-efficient, dual-phase electrochemical platform that integrates solution-phase CRISPR/Cas12a reactions with a tetrahedral DNA nanostructure (TDN)–based interfacial gating mechanism for detecting circulating microRNAs. We found that TDNs extended with a thiolated blocking fragment (BF) impose a pronounced steric gating effect at the electrode–solution interface: the large, rigid TDN architecture prevents intact BF-linked structures from approaching the gold electrode, whereas smaller Cas12a-generated thiolated fragments readily assemble via Au–S interactions. This size-dependent gating transition switches the electrode from an HRP-accessible to an HRP-blocking state, producing a sharply resolved signal decrease that faithfully maps solution-phase cleavage onto an electrochemical readout. Using a target-competitive binding strategy to generate the activator DNA, this interfacial gating mechanism was developed into a robust dual-phase sensing platform driven by CRISPR/Cas12a. Because target recognition and Cas12a-mediated cleavage occur entirely in solution and only the final reaction products contact the electrode, the architecture retains high homogeneous reaction efficiency while avoiding diffusion-governed limitations and eliminating probe immobilization. The 10 µL reaction volume enables substantial reagent savings and strong compatibility with complex matrices. The assay achieves a detection limit of 4.1 aM and accurately quantifies renal cancer–associated circulating miRNA-21 in peripheral blood, yielding results fully consistent with RT-qPCR.

PURPOSE:Immediate urinary continence (UC) recovery following Retzius-sparing robot-assisted radical prostatectomy (RS-RARP) remains highly variable, highlighting the need for reliable preoperative prediction. We aimed to develop and validate models to identify patients likely to achieve immediate UC recovery following RS-RARP. MATERIALS AND METHODS:A total of 580 prostate cancer patients who underwent RS-RARP from four medical centers were assigned to a training set (n=348), an internal validation set (n=103) and an external validation set (n=129). Independent predictors were identified through univariate analysis and LASSO regression. A nomogram was constructed using multivariate logistic regression. Its performance was evaluated with receiver operating characteristic (ROC) curve, calibration curves, and decision curve analysis. RESULTS:Immediate UC recovery was observed in 84.5% (294/348) of patients in the training cohort, 80.6% (83/103) in the internal validation cohort, and 81.4% (105/129) in the external validation cohort, respectively. Multivariate analysis identified membranous urethral length (MUL) (OR=1.23, P=0.029) and urethral curvature (OR=2.84, P<0.001) as independent predictors, while prostate volume (PV) (OR=0.84, P <0.001) as a protective factor. The nomogram integrating MUL, PV, and urethral curvature demonstrated superior predictive accuracy, with an AUC of 0.87 (95% CI, 0.83-0.91) in the training cohort. The bootstrap-corrected calibration slope was 0.96, and the Brier score was 0.08. Calibration curves and decision curve analysis confirmed the predictive accuracy and clinical utility of the nomogram. CONCLUSIONS:Our study introduces a novel quantitative method for assessing urethral curvature. The mpMRI-based model, integrating urethral curvature and prostate spatial configuration, offers enhanced predictive accuracy for postoperative immediate UC recovery.

Androgen deprivation therapy (ADT) for prostate cancer (PCa) leads to lineage plasticity in PCa cells, promoting the emergence of androgen receptor-negative neuroendocrine prostate cancer (NEPC). NEPC is a highly aggressive subtype with poor prognosis and limited treatment options. Tumor-associated macrophages (TAMs) contribute to tumor progression through exosome-mediated communication. In our previous study, we analyzed RNA-seq data to identify key genes involved in PCa progression, and the RERG gene emerged as a significant candidate that suppresses neuroendocrine differentiation (NED). In this study, we demonstrate that RERG expression is significantly reduced in CRPC cells and NEPC tissues, and its downregulation activates the Ras/ERK signaling pathway, which plays a crucial role in promoting NED. Additionally, miR-142-5p, transferred from TAMs via exosomes, downregulates RERG expression and activates the Ras/ERK pathway, thereby promoting PCa progression and NED. In vitro, miR-142-5p enhanced PCa cell proliferation, migration, invasion, and NED, while RERG overexpression reversed these effects. In vivo, RERG knockdown significantly promoted tumor growth and NED in a xenograft model. These findings highlight the role of TAM-derived miR-142-5p in regulating NED and suggest that targeting the RERG/Ras/ERK axis may provide a novel therapeutic approach for NEPC.

This study aimed to investigate the imaging characteristics of Microphthalmia Transcription Factor (MIT) Family Translocation Renal Cell Carcinoma (MIT-RCC), assess the relationship between CT-based radiomics and MIT-RCC, and develop a predictive model to improve preoperative non-invasive diagnosis. A retrospective analysis was conducted using preoperative computed tomography (CT) images and clinical data from 746 RCC patients (77 MIT-RCC and 669 other RCC subtypes) across multiple centers. Regions of interest (ROIs) in both tumor and normal renal parenchymal during non-contrast and arterial phases were manually segmented using ITK-SNAP. Radiomics features were extracted via Python, and the Least Absolute Shrinkage and Selection Operator (LASSO) method was applied for feature selection. A combined model incorporating radiomics scores and clinical predictors (age, stage) was developed using logistic regression and visualized as a nomogram. Patients from two centers formed the training cohort (n = 539), and three other centers served as the validation cohort (n = 207). From 1316 features per ROI, 22 tumor and 24 normal renal parenchymal features were selected. The nomogram integrating Tumor_Radscores, Kidney_Radscores, age, and clinical stage showed strong predictive performance in the training cohort (area under the curve [AUC] = 0.952, 95

PURPOSE:We designed and synthesized a novel compound targeting C-X-C motif chemokine receptor 4 (CXCR4), JH120061. Using clinically established Pentixafor and Pentixather as references, we evaluated the potential of [68Ga]Ga/[177Lu]Lu-JH120061 in a series of preclinical and clinical studies. PATIENTS AND METHODS:Preclinical studies of [68Ga]Ga/[177Lu]Lu-JH120061 were conducted on CXCR4-expressing cell lines (HT1080-hCXCR4) and HT1080-hCXCR4 tumor-bearing mice. A head-to-head comparison of [68Ga]Ga-JH120061 with [68Ga]Ga-Pentixafor for PET/CT was conducted in patients with multiple myeloma (n = 5) and renal masses (n = 5). An expanded cohort of 53 patients with renal masses underwent [68Ga]Ga-JH120061 PET/CT to assess its performance in identifying renal malignancy. RESULTS:Preclinical studies revealed that JH120061 demonstrated high binding affinity for CXCR4, promising cellular uptake and retention. In a clinical study, [68Ga]Ga-JH120061 PET/CT detected more malignant lesions than [68Ga]Ga-Pentixafor (94 vs. 81, P = 0.031) and showed significantly higher tumor uptake [maximum standardized uptake value (SUVmax) 22.3 ± 12.9 vs. 9.4 ± 6.5, P < 0.001, at 60 minutes]. Furthermore, [68Ga]Ga-JH120061 PET/CT exhibited excellent detectability for clear-cell renal cell carcinoma (ccRCC); its tumor uptake was significantly higher than that in non-ccRCC (SUVmax 28.9 ± 11.7 vs. 7.3 ± 2.4, P < 0.001). CONCLUSIONS:This study demonstrated that JH120061 may have excellent affinity for CXCR4. Notably, [68Ga]Ga-JH120061 PET/CT demonstrated a remarkable capability for detecting ccRCC. Future studies should further explore the potential of [68Ga]Ga/[177Lu]Lu-JH120061 in precision theranostics of CXCR4-positive tumors.

Post-traumatic urethral stricture, characterized by hypertrophic scar formation, poses significant therapeutic challenges. Infiltration of neutrophil extracellular traps (NETs) is a prominent feature of post-traumatic urethral microenvironment, but their roles remain unknown. To investigate the mechanisms by which NETs mediate urethral fibrosis and explore potential therapeutic strategies targeting NETs. Single-cell RNA sequencing was performed on urethral tissues from New Zealand rabbits with urethral injury to reveal the cell atlas of post-traumatic inflammatory microenvironment. The level of was assessed in both clinical and animal samples using immunohistochemistry and ELISA. In vitro, NETs were co-cultured with urethral fibroblasts (UFBs) to assess their effects and the mechanism underlying. Additionally, animal models of urethral injury were treated with NETs inhibitors to assess the effects of NETs on urethral scar formation. We constructed a single-cell atlas of the post-traumatic urethral microenvironment and found that NETs infiltration was correlated closely with the development of urethral fibrosis. NETs activated UFBs via the TLR9/NF-κB/Smad3 pathway, promoting collagen synthesis and IL-8 secretion, which further recruited neutrophils, creating a positive feedback loop. In vivo, DNase I and AT791 significantly reduced NET levels, suppressed fibroblast activation, and mitigated collagen deposition, effectively reducing stricture formation. NETs drive post-traumatic urethral fibrosis by activating fibroblasts and amplifying inflammation. Targeting NETs or their uptake via TLR9 inhibition may offer a promising therapeutic strategy for urethral stricture. NETs drive post-traumatic urethral fibrosis by activating fibroblasts and amplifying inflammation. Targeting NETs or their uptake via TLR9 inhibition may offer a promising therapeutic strategy for urethral stricture.

Prostate cancer (PCa) is one of the most common malignancies in men. There is limited data available regarding potential minimally-invasive biomarkers for predicting PCa outcomes and disease monitoring. Here, we investigate the proteomic profile of plasma in 222 patients with PCa and 159 healthy controls. Integrative analyses of the proteome profile and clinical features identified protein networks related to International Society of Urological Pathology (ISUP) grades and prostate-specific antigen (PSA). Proteome-based classification revealed three subtypes, PCa-I, PCa-II, and PCa-III, reflecting distinct clinical prognosis and molecular signatures. We develop a 17-protein panel and established a biochemical recurrence prediction model that effectively predicts biochemical recurrence for patients with PCa, which is better than ISUP grades and pathological stages. Finally, we validate the protein panel by parallel reaction monitoring (PRM) assay in an independent cohort. Collectively, this study portrays the plasma proteomic landscape of PCa cohort and provides a comprehensive resource for further biological and predictive research in PCa.

BACKGROUND:To develop and externally validate CT-based radiomics-clinical nomograms integrating dual-region radiomics features and preoperative-intraoperative clinical factors for predicting early renal function decline in patients with localized renal cell carcinoma (RCC) undergoing partial nephrectomy (PN). METHODS:This multicenter retrospective study included 1440 patients with localized RCC who underwent PN and preoperative contrast-enhanced CT. Radiomics features were extracted from both the tumor and ipsilateral normal renal parenchyma across corticomedullary, nephrographic, and excretory phases. After reproducibility filtering, high-correlation removal, and least absolute shrinkage and selection operator (LASSO) regression, three radiomics signatures were constructed. Independent clinical predictors identified via multivariate logistic regression were combined with radiomics signatures to develop a preoperative nomogram and a preoperative-intraoperative nomogram. Model performance was assessed using ROC analysis, AUC, calibration curves, Hosmer-Lemeshow tests, decision curve analysis (DCA), and 1000-iteration bootstrap validation. External validation was conducted across 10 independent medical centers and an online public dataset (KITS23). RESULTS:Twelve tumor-derived and 8 kidney-derived radiomics features were selected, and the combined radiomics signature demonstrated superior predictive ability across cohorts. Independent predictors included age, diabetes, preoperative eGFR, RENAL score, ischemia time, and the radiomics signature. The preoperative nomogram showed excellent discrimination (AUC = 0.952, 0.909, 0.931, 0.914, 0.926), robust calibration, and favorable DCA performance. Bootstrap analysis confirmed its internal stability (mean AUC = 0.948; 95% CI: 0.927-0.972).The preoperative-intraoperative nomogram further improved discriminative performance (AUC = 0.962, 0.926, 0.947, 0.947, 0.933), with strong calibration and consistent clinical utility across all cohorts. Bootstrap validation also demonstrated excellent internal robustness (mean AUC = 0.960; 95% CI: 0.947-0.990). CONCLUSIONS:Radiomics signatures derived from CT images of both tumor and normal renal tissue, when integrated with clinical parameters, can accurately predict early renal function decline following PN. The proposed nomograms may facilitate individualized risk stratification and optimize surgical decision-making.

Ambient fine particulate matter (PM2.5) is an established environmental health hazard, but its role in clear cell renal cell carcinoma (ccRCC) progression remains incompletely understood. This study integrated epidemiological analysis, mechanistic experiments, transcriptomic profiling, and clinical tissue validation to investigate the association between ambient PM2.5 exposure and ccRCC aggressiveness. Among 1410 patients with pathologically confirmed ccRCC, long-term residential PM2.5 exposure during the 36 months before diagnosis was estimated using geocoded addresses and the Tracking Air Pollution in China dataset. Each 10 μg/m3 increase in PM2.5 exposure was independently associated with advanced AJCC stage (adjusted OR, 1.42; 95% CI, 1.15-1.74) and higher WHO/ISUP grade (adjusted OR, 1.36; 95% CI, 1.12-1.65). Locally collected winter ambient PM2.5 exhibited irregular morphology, heterogeneous aggregation, and complex chemical composition. In vitro and in vivo, PM2.5 enhanced ccRCC cell proliferation, clonogenic growth, migration, invasion, epithelial-mesenchymal transition-like remodeling, and tumor growth. Mechanistically, PM2.5 increased intracellular and mitochondrial ROS, activated PI3K/AKT signaling, enhanced HIF-2α protein stability and nuclear accumulation, and increased HIF-related transcriptional output. Complementary antioxidant, pharmacological, genetic, and rescue experiments supported a functional contribution of the ROS/AKT/HIF-2α axis to PM2.5-associated malignant phenotypes. Transcriptomic profiling identified ROS-responsive genes enriched in oxidative stress, PI3K/AKT signaling, hypoxia response, extracellular matrix organization, and EMT-related pathways. PM2.5 also activated NF-κB-related inflammatory signaling, suggesting an additional ROS-responsive inflammatory response. Clinical ccRCC tissues from patients with higher PM2.5 exposure showed increased HIF-2α and Ki-67 expression together with EMT-related protein alterations. Collectively, these findings indicate that long-term ambient PM2.5 exposure is associated with more aggressive ccRCC and support a model in which oxidative stress contributes, at least in part, to tumor progression through AKT/HIF-2α signaling.

Objective Di-(2-ethylhexyl) phthalate (DEHP), a ubiquitous plasticizer of significant health concern due to its endocrine disrupting effects. The study aims to elucidate underlying molecular mechanisms between DEHP exposure and prostate cancer (PCa). Methods In vitro and in vivo experiments were undertaken to validate the effects of DEHP on the biological function of prostate cancer, and RNA sequencing was performed to elucidate the underlying mechanism. We integrated in silico toxicological assessments and bioinformatics methodologies was employed to investigate the key targets and associated downstream pathways in DEHP-induced PCa progression. Results In vitro and in vivo experiments showed that MEHP promoted the proliferation, migration, and invasion of PCa cells and tumorigenicity, and RNA sequencing revealed that the metabolic pathways, pathways in cancer, PI3K-Akt signaling pathway, MAPK signaling pathway might be potential mechanisms involved in this process. Additionally, six DEHP-related genes were identified and a DEHP-related model was constructed. The data of scRNA indicated that these six DEHP-related genes may contribute in the tumor microenvironment of PCa. The molecular docking analysis revealed DEHP's ability to bind to crucial DEHP -related proteins. Conclusions Our study unveils a novel insight into the function for DEHP in PCa, shedding new light on prediction, assessment, and mitigation of DEHP exposure and PCa progression.

Soft tissue sarcoma is a broad family of mesenchymal malignancies exhibiting remarkable histological diversity. We portray the proteomic landscape of 272 soft tissue sarcomas representing 12 major subtypes. Hierarchical classification finds the similarity of proteomic features between angiosarcoma and epithelial sarcoma, and elevated expression of SHC1 in AS and ES is correlated with poor prognosis. Moreover, proteomic clustering classifies patients of soft tissue sarcoma into 3 proteomic clusters with diverse driven pathways and clinical outcomes. In the proteomic cluster featured with the high cell proliferation rate, APEX1 and NPM1 are found to promote cell proliferation and drive the progression of cancer cells. The classification based on immune signatures defines three immune subtypes with distinctive tumor microenvironments. Further analysis illustrates the potential association between immune evasion markers (PD-L1 and CD80) and tumor metastasis in soft tissue sarcoma. Overall, this analysis uncovers sarcoma-type-specific changes in proteins, providing insights about relationships of soft tissue sarcoma.

The resistance to radiotherapy of prostate cancer is driven by interactions within the tumor microenvironment, particularly between prostate cancer cells and tumor-associated macrophages, however the underlying mechanisms remain poorly understood. In this study, we found that STAT1 enhanced the transcription of critical glycolytic enzymes, leading to an increase in lactate secretion from prostate cancer cells. Then, the lactate was transported to macrophages via the MCT1 transporter, activating the NFκB1 pathway, which subsequently promoted macrophage polarization to the M2 phenotype and activated the transcription of MCP-1. MCP-1 was secreted from macrophages interacted with the CCR2 receptor on prostate cancer cells, thereby activating the JAK/STAT1 pathway, ultimately contributing to the progression of prostate cancer and its resistance to radiotherapy. Taken together, our findings identified a STAT1/lactate/NFκB1/MCP-1 positive feedback mechanism as a driver of prostate cancer progression and resistance to radiotherapy that functioned by interaction to macrophages, which could be potential therapeutic targets for the advanced prostate cancer.

Docetaxel (DTX) is a standard chemotherapy agent for castration-resistant prostate cancer (CRPC); however, DTX resistance remains a major clinical challenge, and the underlying molecular mechanisms are not fully understood. In our study, it was found that OTUB2 was highly expressed in DTX-resistant CRPC and could be served as a key driver of DTX resistance. Mechanistically, OTUB2 stabilizes the m5C reader ALYREF by removing its K48-linked polyubiquitin chains, leading to increased ALYREF protein levels. And then, ALYREF enhances the mRNA stability and expression of ABCG4, thereby promoting ATP-dependent efflux of DTX. Moreover, the expression of OTUB2 mRNA and protein could be regulated by FOXD3-AS1 derived from cancer-associated fibroblasts (CAFs). More importantly, treatment with OTUB2 inhibitor (OTUB2-IN-1) resensitized resistant CRPC to DTX. Together, our findings establish OTUB2 as a novel driver of DTX resistance in CRPC and highlight the role of CAFs-derived FOXD3-AS1 and OTUB2/ALYREF/ABCG4 axis in modulating DTX resistance of CRPC.

Our team previously reported a preclinical study of 2 novel prostate-specific membrane antigen (PSMA)-targeted compounds that incorporate a hypoxia-sensitive nitroimidazole (NI) moiety-6-amino-6-methylperhydro-1,4-diazepinetetraacetic acid-PSMA-093 (AZ-093) and its hypoxia-responsive derivative 6-amino-6-methylperhydro-1,4-diazepinetetraacetic acid-NI-PSMA-093 (AZ-NI-093). This prospective trial aims to evaluate the clinical value of both agents in patients with prostate cancer, specifically exploring the impact of NI on tumor uptake. Methods: Sixty patients were enrolled in this PET/CT imaging study. Thirty patients underwent a head-to-head comparison of [68Ga]Ga-AZ-093 with [68Ga]Ga-PSMA-11 or [68Ga]Ga-PSMA-617, and 30 patients underwent a head-to-head comparison of [68Ga]Ga-AZ-NI-093 with [68Ga]Ga-AZ-093. The number of tumors and SUV for tumors and organs were measured and recorded. SUVmax differences between [68Ga]Ga-AZ-NI-093 and [68Ga]Ga-AZ-093 PET/CT were calculated for further analysis. Immunohistochemical staining for hypoxia-inducible factor 1 was performed on 12 surgical specimens of intraprostatic tumors. Results: All patients tolerated [68Ga]Ga-based PET/CT scans without adverse effects. The initial biodistribution of [68Ga]Ga-AZ-093 and [68Ga]Ga-AZ-NI-093 in humans, as assessed by PET/CT, was comparable to that of other PSMA-targeted radiopharmaceuticals, with respective effective absorbed doses of 0.0128 ± 0.00594 and 0.0160 ± 0.000869 mSv/MBq. [68Ga]Ga-AZ-093 exhibited higher tumor uptake 60 min after injection compared with [68Ga]Ga-PSMA-11 (SUVmax, 22.2 ± 10.8 vs. 20.4 ± 14.8; P = 0.025) and [68Ga]Ga-PSMA-617 (SUVmax, 20.1 ± 12.7 vs. 10.8 ± 8.7; P < 0.001), while maintaining comparable tumor detection rates. [68Ga]Ga-AZ-NI-093 showed further improved tumor uptake relative to [68Ga]Ga-AZ-093 at 60 min (SUVmax, 17.8 ± 14.0 vs. 16.3 ± 11.5; P = 0.009) and 150 min (31.3 ± 18.5 vs. 28.7 ± 17.5; P = 0.003), especially in large-volume, high-grade intraprostatic tumor and metastatic sites. The expression of hypoxia-inducible factor 1 in tumors, as an indicator of hypoxia, demonstrated a significant association with International Society of Urological Pathology grade (r = 0.519; P = 0.038) and difference in SUVmax (r = 0.629; P = 0.023). Conclusion: Both [68Ga]Ga-AZ-093 and [68Ga]Ga-AZ-NI-093 represent promising PSMA-targeted radiopharmaceuticals. [68Ga]Ga-AZ-NI-093 demonstrated potential in targeting hypoxic tumor tissues. Future studies with larger sample sizes are needed to further explore the clinical value of these radiopharmaceuticals.