The Prostate

BackgroundIdentifying metabolites associated with prostate cancer (PC) aggressiveness may elucidate mechanisms underlying disease severity. Doing so for plasma and formalin-fixed, paraffin-embedded (FFPE) tissue could accelerate discovery. In this cross-sectional pilot study, we generated hypotheses for further exploration by assessing associations between plasma metabolites and Gleason score in individuals with PC and evaluating correlations between plasma and FFPE metabolite levels. MethodsWe examined plasma and FFPE samples from 10 individuals with Gleason score 7 (six 3+4, four 4+3) and nine individuals with Gleason score 9 (six 4+5, three 5+4) tumors from a convenience sample of 19 men with PC. We measured the relative abundance of polar metabolites at the time of radical prostatectomy. We used linear models of log2 fold changes to examine plasma metabolite levels relative to pathologic tumor grade. Relationships among metabolite levels measured in plasma and FFPE tumor tissue within individuals across metabolites were examined using Pearson correlations. ResultsAmong 18 plasma metabolites selected a priori because of prior associations with PC aggressiveness, serine (p=0.0051) and ornithine (p=0.036) levels were higher in individuals with Gleason 9 than Gleason 7 PC. After multiple testing correction, however, no associations were statistically significant. The median correlation between levels in plasma and FFPE tumor tissue was 0.45 (range: 0.40-0.53) for the 94 metabolites measured in both biospecimens. ConclusionsPlasma serine and ornithine demonstrated the largest differences between individuals with Gleason 7 and Gleason 9 PC. Metabolite levels in FFPE prostate tissue samples were moderately correlated with plasma levels. Future studies in larger samples are needed to further explore the hypotheses generated by this study.

Background: Previous recommendations on screening for prostate cancer relied on ongoing trials of screening with prostate-specific antigen (PSA), which may have lacked sufficient follow-up duration to fully examine effects on mortality and overdiagnosis. Findings which consider absolute effects by age and screening intensity, along with newer guidance for assessing evidence certainty, may lead to different interpretations. Adding magnetic resonance imaging (MRI) to PSA-based screening has been raised as a way to reduce false positives (FPs) and overdiagnosis. Methods: We systematically searched MEDLINE, Embase, and Central from 2014 to January 28, 2026, for randomized controlled trials (RCTs) and prospective observational studies of: (i) screening versus no screening and (ii) sequential screening with MRI for those with a positive PSA test versus PSA alone among men not known to be at high risk for prostate cancer. Studies on screening with PSA or digital rectal examination (DRE) published pre-2014 were identified from existing systematic reviews and reference lists. Studies on FPs and complications from biopsies after PSA screening did not require a control group. Paired reviewers screened titles/abstracts (assisted with artificial intelligence) and full texts, assessed risk of bias, and extracted data, by age when available. We pooled data when suitable using random-effects models, investigated heterogeneity, and assessed the certainty of evidence using GRADE with conclusions of effects based on decision thresholds based on absolute effect sizes. Results: Across both questions, we included 15 RCTs (N=856,000; 8 sites of ERSPC considered separate trials) and 8 observational studies (N=56,122). At 20 years, among 1000 men who underwent repeated PSA-based screening every 2-4 years starting from age 55-69 (mean 62), there is likely a reduction in prostate-cancer mortality ([≥]2 fewer) and metastatic cancer incidence ([≥]6 fewer), at the expense of prostate-cancer overdiagnosis ([≥]24 cases) and FPs ([≥]150 cases) (all moderate certainty). If screening starts at age 50-54 or age 55, the benefits are probably smaller (e.g., 1 vs. 2 fewer prostate-cancer related deaths) with similar harms. Adding DRE or screening with PSA annually does not add benefit. One round of PSA screening or starting screening later at age 70-74 may not offer any important benefit or harm (low to moderate certainty), and any benefit from screening primarily with DRE was not shown. Compared with PSA alone, sequential screening with PSA followed by MRI reduces FPs ([≥]33 fewer) and overdiagnosis (via [≥]10 fewer diagnoses of clinically insignificant [e.g., Gleason 6] cancers without impacting detection of clinically significant cancers) (moderate to high certainty), though findings were limited to one round of screening without long-term follow-up or measurement of mortality. Interpretation: This review provides clinicians and other interest holders with anticipated absolute effects by age, and assessments of certainty across critical and important outcomes and with approximately two decades of follow-up. Findings apply to a general population and may differ for specific groups. Results for most critical outcomes, both benefits and harms, exceeded thresholds for clinically important effect sizes, thereby demonstrating the complexity of guideline developers' and patients' decision-making regarding screening trade-offs. Findings about adding MRI for those with a positive PSA test were limited and would require additional consideration of costs, infrastructure, expertise, and equity. Protocol registration: PROSPERO - CRD420250651056.

BackgroundDespite extensive cataloging of carcinogenic exposures by the International Agency for Research on Cancer (IARC) and pharmacogenomic variation by resources such as PharmVar and CPIC, few platforms unify exposure, metabolic activation and detoxification, DNA damage, and genetic annotation within a single interactive visualization framework. This gap limits systematic evaluation of gene-environment interactions in cancer risk assessment. MethodsWe developed the Carcino-Genomic Knowledge Graph, ExposoGraph, an interactive knowledge-graph platform for carcinogen metabolism and DNA damage pathways. The reference graph integrates curated data and annotations from IARC, KEGG, PharmVar, CPIC, CTD, and supporting literature/resources. The current reference graph contains 96 nodes across 5 entity types (Carcinogens, Enzymes, Metabolites, DNA Adducts, and Pathways) and 102 edges across 6 relationship types (activates, detoxifies, transports, forms adduct, repairs, and pathway). ResultsThe first-generation reference graph captures metabolic activation and detoxification pathways for 9 carcinogen classes spanning 15 index carcinogens. It represents 36 enzymes across Phase I activation (n=14), Phase II conjugation and detoxification (n=14), Phase III transport (n=3), and DNA repair (n=5). Interactive exploration supports carcinogen-class filtering, node- and edge-type filtering, metadata-based search, and detailed hover/detail views with provenance and pharmacogenomic annotations. The androgen branch highlights cross-pathway connectivity by linking androgen metabolism to estrogen quinone formation and DNA adduct generation through CYP19A1-mediated aromatization and downstream catechol estrogen chemistry. In the optional androgen-focused extension, additional receptor, tissue, and variant context further connects this branch to androgen receptor signaling and genotype-specific annotations. ConclusionsExposoGraph provides a first-generation integrated, interactive framework linking carcinogenic exposures to metabolic fates and genetic modulators. The platform supports hypothesis generation for gene-environment interaction studies and may inform future individualized risk modeling, while remaining a research-use framework rather than a clinically validated risk-assessment tool.

Whole-genome sequencing (WGS) enables comprehensive analysis of tumour genomes, but its use in formalin-fixed paraffin-embedded (FFPE) samples is limited by DNA fragmentation and low yields. Whole-genome amplification (WGA) methods such as multiple displacement amplification (MDA) can boost DNA availability but distort copy-number alteration (CNA) profiles. DNA ligation-mediated MDA (DLMDA) mitigates this bias by reconstituting fragmented templates, yet its performance in FFPE-derived DNA remains uncertain. We compared paired DLMDA pre-amplified (2h, 8h) and non-pre-amplified FFPE prostate tumour samples from 22 archival blocks (5, 15 and 20 years old). DLMDA increased DNA yield by 42- to 86-fold, with global CNA patterns largely preserved. However, DLMDA significantly reduced the number of detected CNA deletions and amplifications. These effects were independent of both block age and reaction time. CNA dropouts were randomly distributed across the genome, indicating that DLMDA does not introduce regional bias. Our results show that DLMDA enables robust DNA yield recovery and avoids false-positive CNA artefacts, but at the cost of reduced CNA sensitivity. While suitable for CNA screening pipelines through WGS, further improvements are required to minimise the false-negative risk and improve the techniques sensitivity for FFPE-based genomics.

Cannabis (marijuana) is the most widely used recreational drug in the USA accounting for about 62 million users in 2024. Among cannabis users, 26% are of prime reproductive age (18-25 years). Delta-9 tetrahydrocannabinol (THC) is the principal psychoactive component of cannabis and has been detected in human seminal fluids. Although abundant evidence indicates adverse effects of THC exposure on spermatogenesis in different species, acute effects of THC on postejaculatory sperm including fertilization potential and subsequent carryover effects on embryo development are largely unknown. The present study was designed to provide missing information on structural and mechanistic effects of THC exposure to postejaculatory sperm function by evaluating sperm indices often overlooked or masked during clinical evaluation. A bovine embryo continuum model was employed to determine effects of THC on sperm structure, kinematics, bioenergetics, and binding mechanisms. Effects of THC on the sperm genomic and epigenomic landscape were determined, complemented by paternal carry over effects on embryo development as a human translational model to elucidate paternal effects on future development, and to mirror sperm exposure during transport within the female reproductive tract. Cryopreserved bovine sperm from three bulls were independently exposed to physiologically relevant concentrations of THC (0 and 32nM, n = 2 individual replicates/bull) for 24 h under non-capacitating conditions at 25{degrees}C followed by quantification of sperm kinematics at 37{degrees}C. Samples of THC-exposed sperm and vehicle-control (0.1% DMSO) were collected in replicate following immediate addition of THC (0 h) and again at 24 h. DNA damage, acrosome integrity, bioenergetics, changes to DNA methylation and embryo development were quantified. Data were analyzed by logistic regression with a generalized linear mixed effect model. Computer-assisted sperm assessment revealed a reduction in progressive motility of THC-exposed sperm after 24 h while other parameters were not affected. Acrosome integrity as determined by flowcytometric analysis with FITC-PSA was severely compromised in THC-exposed sperm (P [≤] 0.05), despite no detectable difference in capacitation status using merocyanine staining. Similarly, DNA integrity as determined by TUNEL assay was significantly impaired after 24 h of THC exposure (P [≤] 0.05). Mechanistic effects of THC were explored through characterization of the transmembrane G-protein coupled cannabinoid 1 receptor (CB1). CB1 is expressed in the post-acrosomal region and its abundance decreased as compared to unexposed sperm. Alterations to the methylation landscape of sperm were then determined after 24 h of THC exposure through whole-genome Enzymatic Methyl Sequencing. PCA analysis indicated that sperm from different males formed distinct clusters, implying individual differences among bulls, while the effects of THC exposure produced tighter clusters. Paternal carryover effects on embryos derived by in vitro fertilization from THC exposed sperm had reduced 2-cell cleavage, 8-16 cell morula development, and reduced blastocyst development compared to unexposed sperm (46% vs. 33%). In conclusion, post-ejaculatory mammalian sperm exposure to THC compromises acrosome integrity, induces DNA damage, changes the sperm methylome, and reduces developmental potential. Collectively, these data implicate new considerations for recreational and clinical use of cannabis that impact cellular and molecular mechanisms important for sperm function with detrimental consequences for gamete interaction and embryo development.

Prostate cancer (PCa) is the second most common cancer and cause of cancer death in American men. Existing risk prediction methods have limited accuracy and reproducibility, resulting in difficulty in predicting disease severity. We demonstrate the development and external validation of an automated multimodal artificial intelligence algorithm using biparametric MRI (bpMRI) and clinical covariates for predicting biochemical recurrence (BCR) after radical prostatectomy (RP) in PCa patients. Development cohort included 80% of patients from center 1 (n = 240) who underwent prostate MRI prior to RP between January 2008 and December 2018 with a minimum of two years of follow-up after RP. Test cohort included the remaining 20% of center 1 patients (n = 71), and the external validation cohort from center 2 (n = 168). Center 2 patients included those who underwent prostate MRI and RP between January 2015 and December 2024 with a minimum of two years of follow-up. Clinical comparisons were CAPRA-S (center 1) and ISUP grade group from post-RP biopsy (center 2). Models developed were a clinical model (M0), an automated clinical model (M1), a radiomics model (M2), and a multimodal model (M3). Clinical variables (M0) included PSA, age, primary Gleason, and ISUP grade group. Automated clinical variables (M1 and M3) included PSA and age. Radiomics features (M2 and M3) were extracted from bpMRI using a lesion detection algorithm. Accuracy, sensitivity, specificity, and AUC were calculated, and log-rank tests compared BCR-free survival to assess the models ability to discriminate relative to clinical standards. Intermediate-risk groups were also assessed. The multimodal model (M3) had the highest AUC across test sets (combined: 0.71; center 1: 0.70; center 2: 0.75) and was the only model to significantly differentiate BCR-free survival outcomes in intermediate-risk groups across both centers (p < 0.05). This automated multimodal model leveraging radiomics and clinical covariates can predict BCR after RP, approaches clinical gold standards, and may enhance imaging-based prognostication following further validation.

Microbial steroid metabolism represents an underappreciated extension of the vertebrate endocrine system, with growing evidence that host-associated microbes contribute to the diversity and bioavailability of sex steroids within human tissues. Emerging studies have linked microbial androgen metabolism to urinary microbiome composition and to resistance to androgen deprivation therapy (ADT) in prostate cancer. While microbial pathways capable of converting steroid precursors such as cortisol to androgens, via the steroid-17,20-desmolase pathway, such as DesG-mediated interconversion of androstenedione to testosterone have been reported, the diversity of enzymes mediating downstream androgen interconversion remains incompletely defined. Here, we investigate the androgen-forming capabilities of anaerobic bacteria from the male genitourinary microbiome, focusing on NADPH-dependent 17{beta}-hydroxysteroid dehydrogenases (17{beta}-HSDHs) that catalyze interconversion of androstenedione and testosterone. We isolated androgen-forming bacterial strains from human male urine and identified a previously uncharacterized 17{beta}-HSDH encoded by Peptoniphilus obesi, demonstrated that this enzyme catalyzes the NADPH-dependent reduction of androstenedione to testosterone and the reverse oxidation reaction. Sequence similarity searches further identified a homologous 17{beta}-HSDH in Anaerococcus, which was synthesized and functionally validated, revealing conserved activity despite low sequence identity to the previously characterized urinary tract enzyme DesG. The enzymes were found to have broad substrate specificity for C19 and C18 17keto- and 17{beta}-hydroxysteroids. Together, these findings expand the known diversity of microbial 17{beta}-HSDHs and identify previously unrecognized androgen-forming activities within the genitourinary microbiome. ImportanceMicrobial steroid-transforming pathways may provide a mechanism by which commensal anaerobes contribute to androgen availability in the genitourinary tract. By identifying novel 17{beta}-hydroxysteroid dehydrogenases from Peptoniphilus and Anaerococcus, genera repeatedly associated with prostate cancer, this study provides mechanistic insight into how microbial steroid metabolism may influence hormone-driven disease.

Ionizing radiation can be an effective therapy for prostate cancer. Unfortunately, however, more aggressive prostate cancers such as neuroendocrine prostate cancer (NEPC) are often radiation resistant, which contributes to their high degree of morbidity and mortality. In this study, we used an unbiased approach to identify novel mechanisms that contribute to resistance to radiation and that are associated with neuroendocrine differentiation. Specifically, we compared the expression of cell surface proteins by mass spectrometry in prostate cancer cell lines that had been either untreated or treated with radiation to induce resistance, a process that also promotes neuroendocrine differentiation. Among the proteins identified by this screen, we focused on folate receptor (FR) because of its known biological functions and the fact that it is a validated therapeutic target. Our data reveal that FR has a causal role in enabling prostate cancer cells to resist radiation. Importantly, we also demonstrate that the expression of FR is regulated by HIF-1, which also has a causal role in radiation resistance and neuroendocrine differentiation. Given that the ability of cells to resist damage and death in response to ionizing radiation depends largely on their ability to buffer the substantial increase in reactive oxygen species (ROS) that is generated by radiation, we also demonstrate that the folate-FR axis promotes radiation resistance by sustaining intracellular glutathione levels that buffer this increase in ROS. In summary, the data reported here highlight a novel role for FR in resistance to ionizing radiation that is intimately associated with the hypoxic microenvironment of NEPC and the ability of the folate-FRa axis to maintain redox homeostasis.

BackgroundWilms tumour (WT) relapse occurs more frequently in patients with blastemal-type WTs. The presence of cancer stem cells (CSCs) is linked to tumour survival and relapse, and CSCs may be found in greater numbers in blastemal cell foci. CSC-associated phenotypes have been described in untreated WT, but their persistence, organisation and relevance after neoadjuvant chemotherapy is unknown. MethodsWe analysed 23 formalin-fixed paraffin-embedded blocks from 18 chemotherapy-treated patients where WTs were enriched for viable blastema, using human fetal kidney as developmental control. Immunohistochemistry and -fluorescence analysis determined progenitor (PAX2, SIX2, CITED1) and CSC-associated (NCAM, ALDH1, CD133) marker expression. We qualitatively and semi-quantitatively evaluated spatial expression patterns and co-localisation across tumour compartments. ResultsPAX2 and SIX2 were co-expressed in blastema in most cases (15/18), with PAX2 expression higher at the periphery of blastemal foci and SIX2 expression found uniformly in central aspects. CITED1 expression was also associated with SIX2 in blastema tissues (14/18). NCAM was blastema-enriched (15/18) with higher central intensity, frequently adjacent to PAX2-expressing peripheral zones. ALDH1 expression was present across blastema and epithelium while NCAM-, ALDH1-double-positive cells were rarely observed (4/18). CD133 expression was less commonly seen (2/18), localising near epithelial/nephrogenic structures. ConclusionsAfter neoadjuvant chemotherapy, WT blastema retained overlapping but non-identical progenitor/CSC-associated marker landscapes with reproducible peripheral-centre gradients. These spatial arrangements suggest a blastemal niche for CSCs that may sustain a therapy-resistant state. Our analysis provides the foundation for future functional validation and molecular profiling to define key lineage relationships and therapeutic vulnerabilities in post-chemotherapy WT. [250/250 words]

BACKGROUND: Prostate cancer (PrCa) is the most commonly diagnosed cancer in men in many countries and is the most heritable of the common cancers. Precision medicine approaches to disease management are not routinely available to most men, yet we know that germline genetic testing can help identify those at high-risk of developing advanced or lethal disease and can influence selection of therapeutics. An integral part of healthcare delivery design is the inclusion of patients/consumers in the development of frameworks for managing health interventions that are tailored to meet their needs. METHODS: In Phase I, we undertook focus group discussions with men previously diagnosed with PrCa (n=20), to determine their opinions, perceptions and expectations of germline genetic testing for PrCa. Focus groups were tape-recorded, transcribed verbatim, coded and then thematically analysed using NVivo. In Phase II, themes were then used to design and development a Precision Medicine in Prostate Cancer Information Toolkit, which was reviewed by patients (n=14), their carers/family members (n=4) and healthcare providers (n=14). RESULTS: In Phase I, knowledge about precision medicine and genetic testing was generally low. The strongest motivation for undertaking testing was to identify family members' risk levels (n=7), and the biggest concern pertained to insurance discrimination (n=5). Phase II data revealed that generally healthcare providers (n=8) found the purpose of the toolkit to be clearer than patients (n=5). Though, patients found the task of imagining the usefulness of the toolkit at the time of diagnosis or beforehand when assessing genetic risk, quite difficult. Participants highlighted that information regarding life insurance, implications for their family and costs associated with testing were of concern. CONCLUSIONS: This study has revealed critical knowledge gaps, preferred communication/support needs, and concerns/risks associated with germline genetic testing in PrCa. Concerns pertaining to family members and insurance discrimination are obvious topics that need to be addressed. Our toolkit may be helpful in addressing knowledge gaps, but further testing is needed to ensure its accessibility across literary and cultural contexts.

BackgroundWithin the kidney, (pro) renin receptor (PRR) is abundantly expressed in the collecting duct (CD) and modulate physiological and pathophysiological processes. We have recently reported that activation of CD PRR mediates obstructive renal fibrosis in a mouse model of unilateral ureteral obstruction (UUO). The current study addresses the underlying mechanisms by examining the profibrotic pathway mediated by soluble PRR (sPRR)-dependent alternative macrophage activation. MethodsWe performed UUO or sham surgery on mice with CD-specific deletion of PRR (CD PRR KO) and floxed controls. After 7 days, we assessed fibrosis-related parameters, inflammatory cytokines, M1/M2 macrophage markers, other gene expression markers of kidney injury, and the concentration of plasma sPRR. We also administered vehicle or site-1 protease (S1P) inhibitor PF-429242 (PF) to C57BL/6 mice with UUO to determine the role of sPRR. Experiments were performed in vitro to examine the mechanism of sPRR-His-mediated macrophage M2 polarization and activation of potential target genes in bone-marrow-derived macrophages (BMDMs). ResultsCompared with the floxed control, CD PRR KO decreased macrophage accumulation, M2 polarization, and Yap/Taz expression while improving renal fibrosis and suppressing plasma sPRR levels following UUO. In BMDMs, sPRR-His treatment promoted macrophage M2 polarization, fibrosis, and Yap/Taz expression, which was dependent on angiotensin type 1 receptor (AT1R). ConclusionCD PRR-derived sPRR acts via ATR to promote macrophage M2 polarization and stimulates the AT1R/Yap/Taz axis, which leads to renal fibrosis during UUO.

Although several ancillary tests are available in limited laboratories, diagnosis of microphthalmia (MiT)/TFE family translocation renal cell carcinoma (tRCC) could be challenging due to diverse and overlapping tumor morphology and the lack of reliable biomarkers. GPNMB has been recently identified as a diagnostic marker for various renal neoplasms with FLCN/TSC/mTOR-TFE alterations. However, the sensitivity and specificity of GPNMB immunostain are suboptimal and the result interpretation in ambiguous cases could be difficult. To search additional biomarkers that could improve the screening sensitivity and predict genetic aberrations in FLCN/TSC/mTOR-TFE pathway in renal tumors, we performed bioinformatic analysis of publicly available cancer databases and found GPR143, a transmembrane protein regulated by MiT transcription factors, was highly expressed in a subset of renal cell carcinomas (RCCs). In two the Cancer Genome Atlas (TCGA) kidney cancer cohorts, RCCs with high levels of GPR143 expression were enriched for renal neoplasms with FLCN/TSC/mTOR-TFE alterations. Similar to GPNMB labeling, GPR143 immunostain was positive in the majority of tRCC cases and renal tumors with FLCN/TSC/mTOR alterations, suggesting that GPR143 could function as another surrogate marker for FLCN/TSC/mTOR-TFE alterations in certain renal tumors. Interestingly, despite the concordant GPR143 and GPNMB immunoreactivity in most renal neoplasms with FLCN/TSC/mTOR-TFE alterations, diffuse GPR143 immunostain was observed in some cases with negative or focal GPNMB labeling. Taken together, our results indicate GPR143 could serve as a useful adjunct marker to improve the sensitivity for screening renal tumors with FLCN/TSC/mTOR-TFE alterations.

Maternal pancreatic {beta}-cells undergo functional and structural changes to adapt to increased metabolic demands during pregnancy. Lactogen signaling via the prolactin receptor (PRLR) contributes to these adaptations by increasing {beta}-cell mass, insulin transcription and glucose-stimulated insulin secretion[1-4]. In other lactogen-responsive tissues such as the mammary glands and specific hypothalamic nuclei, gestation induces epigenetic changes, some of which persist long after birth[5, 6]. We have previously found that prolactin treatment in islets regulates the expression of epigenetic modifiers[7, 8]. However, whether lactogen signaling in {beta}-cells mediates epigenetic changes to regulate chromatin accessibility has not been examined. Therefore, our objective was to determine whether PRLR signaling alters chromatin accessibility of {beta}-cells to facilitate transcriptional regulation. Using single-cell ATAC-sequencing, we identified differentially accessible regions (DARs) in {beta}-cells which had 718 overrepresented motifs following prolactin treatment of murine islets. Validating this approach, these included motifs bound by established PRLR signaling effectors such as the STAT family of transcription factors (TFs). Using RNA-sequencing we identified transcriptional changes in 41 TFs whose motifs were overrepresented in DARs, including several previously linked to PRLR signaling within {beta}-cells, including Myc, Mafb and Esr1. Importantly, we also identified TFs not previously associated with PRLR signaling, including OVOL2 an established regulator of epigenetic landscape within cells. OVOL2 is a transcription factor involved in EMT inhibition and energy homeostasis with unknown roles in pancreatic {beta}-cells. Here, we establish that OVOL2 acts as a negative regulator of lactogen-dependent effects on {beta}-cell proliferation, establishing a novel regulator of PRLR signaling.

Prostate cancer progression is characterized by dysregulated lipid metabolism, with fatty acid synthase (FASN), the rate-limiting step in de novo lipogenesis (DNL), resulting in significant accumulation of saturated lipids. Here, we investigate whether pharmacologic FASN inhibition creates a metabolic state that increases reliance on exogenous polyunsaturated fatty acids (PUFAs). Inhibition of FASN profoundly alters membrane phospholipid composition, driving compensatory incorporation of PUFAs into membrane phospholipids, thus increasing susceptibility to lipid peroxidation and oxidative damage. Combined FASN inhibition and PUFA exposure increased reactive oxygen species, induced mitochondrial hyperpolarization, and enhanced lipid peroxidation in both hormone-sensitive and castration-resistant prostate cancer models. Marked inhibition of human and murine prostate cancer organoids is achieved ex vivo. In genetically engineered, DNL-reliant Hi-Myc mice, a diet enriched in PUFAs significantly inhibited invasive carcinoma compared to a saturated fat-enriched diet. Environmental PUFAs modulate and enhance the therapeutic efficacy of FASN-targeted strategies. These findings set the stage for pharmacologic and dietary intervention in prostate cancer patients.

BO_SCPLOWACKGROUNDC_SCPLOWThere is increasing momentum behind the clinical implementation of AI-based software for image analysis in digital pathology. As regulations, standards, and national approaches to the clinical use of AI continue to develop, the marketplace of AI products is expanding and evolving - presenting pathologists with a multitude of devices that offer the potential to improve pathology services. MO_SCPLOWETHODSC_SCPLOWTo maintain pace with this changing AI device landscape, we conducted a comprehensive search for, and analysis of, commercial AI products for image analysis in digital pathology. This included CE-marked and Research Use Only (RUO) products using images with histological stains (e.g., H&E) or immunohistochemical (IHC) labelling. Product information and published clinical validation studies were assessed, to understand the quality of supporting evidence on available products, and product details were compiled into a public register: https://osf.io/gb84r/overview. RO_SCPLOWESULTSC_SCPLOWIn total, we identified and assessed 90 CE-marked and 227 RUO AI products. We found that AI products for cancer detection in prostate and breast pathology comprised a substantial portion of the marketplace for H&E image analysis, while IHC products were almost exclusively for use in breast cancer. Clinical validation studies on these products have steadily increased; however, we found that published studies were only available for just over half of H&E products and just over a quarter of IHC products. For CE-marked products, the dataset quality and diversity for AI model performance validation was highly variable, and particularly limited for IHC products. Furthermore, only a limited number of products included studies that assessed measures of clinical utility. CO_SCPLOWONCLUSIONC_SCPLOWAs clinical deployment of AI products for image analysis in histopathology grows, there is a need for transparency, rigorous validation, and clear evidence supporting clinical utility and cost-effectiveness. Independent scrutiny of the expanding offering of AI products provides insight into the opportunities and shortcomings in this domain.

Glioblastoma Multiforme (GBM) is a highly aggressive brain cancer characterized by rapid proliferation and extensive remodeling of the extracellular matrix (ECM), leading to progressive tissue stiffening. Although ECM stiffness is known to promote GBM progression, the molecular mechanisms linking mechanical cues to tumor growth remain insufficiently defined. In this study, transcriptomic comparison of GBM tumors and non-neoplastic brain tissue revealed coordinated upregulation of cell cycle regulators and matrisome-associated genes, with survivin (BIRC5) identified as a central node linking proliferative signaling and ECM remodeling networks. Analysis of GBM patient specimens further showed strong nuclear survivin expression in regions with elevated collagen deposition. To directly evaluate stiffness-dependent regulation of survivin, GBM cells were cultured on fibronectin-infused hydrogels with tunable stiffness. Stiff matrices increased survivin expression along with cyclin D1 and cyclin A, consistent with increased cell cycle progression. Pharmacologic inhibition or siRNA-mediated suppression of survivin reduced stiffness-induced proliferation and attenuated expression of matrisome components, including collagens and lysyl oxidase. These findings indicate that survivin functions as a mechanosensitive regulator that coordinates cell cycle progression with ECM production in stiff tumor microenvironments. Collectively, this study identifies survivin as a key mediator linking ECM stiffness to GBM growth and matrisome remodeling. Targeting survivin and its effectors may offer a mechanosensitive strategy to limit GBM growth.

Natural extracts could improve sperm storage and artificial insemination (AI). This study, for the first time, evaluates the suitability of a blueberry extract (Vaccinium corymbosum) obtained from pomace using a sustainable methodology as a supplement for bull semen extenders. Cryopreserved semen doses from eight bulls were combined in 9 pools (3 bulls/pool), supplemented with 0%, 1%, 5%, or 10% extract, and incubated up to 5 h at 38 {degrees}C. Motility was assessed hourly using OpenCASA, and the effects of treatment and time were evaluated using linear mixed-effects models. Motility was significantly better preserved with 1% extract (total and progressive motility, improved linear velocity and linearities, and decreased BCF and fractal dimension, related to hyperactivation). The effect of 5% was overall positive, but it was below 1%, whereas 10% mostly showed a negative effect. These results show that this natural extract could safely supplement bull semen extenders at least between 1% to 5%, and even help improve sperm motility. Therefore, this extract offers an opportunity to enhance cattle semen extenders using a sustainable approach, potentially improving reproductive outcomes.

BackgroundKidney cell lines are widely used to model kidney physiology and disease; however, their gene expression profiles may differ from primary cells due to immortalization, culture conditions, or experimental treatments. Determining whether a cell line resembles its native cell type is critical for interpreting in vitro findings. We developed a transcriptome-based approach that matches bulk RNA-seq data from kidney cell lines, primary cells, or tissues to reference cell types derived from single-cell RNA-seq (scRNA-seq) datasets. MethodsReference transcriptomic profiles were generated from two human and two murine kidney scRNA-seq datasets by pseudobulk aggregation. Bulk RNA-seq data from microdissected kidney tissue, non-kidney negative controls, and kidney cell lines were matched to these references using three statistical similarity measures (Spearman correlation, Euclidean distance, Poisson distance) and three machine learning classifiers (Random Forest, XGBoost, TabPFN). Each was assessed with global gene expression, curated kidney marker gene lists, and the most variable genes. Matching accuracy was evaluated through a three-step validation strategy: within-dataset matching, cross-reference comparison, and validation against primary kidney tissue and negative controls. ResultsGene expression rank-based Spearman correlation and TabPFN, a foundation model for tabular data, emerged as the most accurate and specific approaches, particularly with curated kidney marker gene lists. Both methods correctly identified microdissected kidney tubule segments and were robust against non-kidney negative controls. Applied to commonly used kidney cell lines, OK cells retained proximal tubule identity, particularly under shear stress, while other proximal tubule lines (HK-2, HKC-8, HKC-11) showed inconsistent matching. Collecting duct-derived mIMCD-3 maintained stable similarity across passages, culture conditions, and genetic modifications. ConclusionWe provide two complementary implementations: CellMatchR, an accessible web-based tool using Spearman correlation for routine use, and comprehensive scripts for TabPFN-based matching (link will be added after peer reviewed publication). Together, these resources enable researchers to make informed decisions about kidney cell culture model selection, interpretation, and stability. Translational StatementKidney cell lines are fundamental tools in nephrology research, yet their transcriptomic similarity to native cell types is rarely validated systematically. We demonstrate that combining bulk RNA-seq data with single-cell reference datasets enables robust assessment of cell line identity using gene expression-rank-based correlation and machine learning approaches. By providing a comprehensive evaluation of matching methods, curated kidney marker gene lists, and reference datasets, our study serves as both a practical resource and a methodological framework for the kidney research community, facilitating informed selection of cell culture models, quality control of experimental conditions, developing new experimental cell culture models, and more reliable translation of in vitro findings to kidney physiology and disease.

Secretory breast carcinoma (SBC) is typically indolent, yet mechanisms underlying aggressiveness and therapeutic resistance to tropomyosin receptor kinase inhibitors (TRKi) remain unclear. Autopsy-based longitudinal multi-organ high-dimensional profiling of metastatic TRKi-resistant SBC demonstrated histopathological heterogeneity, including secretory and squamous components, arising from a shared clonal origin. Integrated genomic and transcriptomic analyses revealed hierarchical transcriptional rewiring consistent with a lineage-plastic state, suggesting a potential link to tumor aggressiveness and therapeutic resistance.

RNF43 is best known for removing the Wnt-receptor complex from the cell surface, thereby maintaining Wnt-signaling at minimal essential levels. Recent studies reported that RNF43-mutant colorectal cancers carrying the common BRAFV600E mutation, respond more effectively to combined BRAF/EGFR inhibition. To determine whether RNF43 directly regulates EGFR or BRAF protein abundance, multiple pancreatic and colorectal cancer cell line models were generated in which RNF43 was knocked out, repaired, or stably overexpressed. Total and cell surface EGFR levels, as well as endogenous BRAF expression, were quantified. Across all models, no consistent evidence emerges that RNF43 modulates endogenous EGFR or BRAF levels. R-spondins likewise fail to alter EGFR levels or internalization. Notably, elevated EGFR expression observed in a subset of RNF43 knockout clones is induced by unintended CRISPR/Cas9 vector integration rather than the absence of RNF43 itself, highlighting a previously underappreciated artefact that can confound interpretations of EGFR regulation in genome edited lines. Overall, the data argue against a direct and general role for RNF43 in controlling EGFR or BRAF protein abundance, contradicting recent reports that propose degradation of these targets. Further studies are required to resolve these discrepancies and clarify the mechanistic basis underlying these conflicting observations.