British Journal of Cancer

BackgroundMesonephric-like adenocarcinoma has been recently classified as a rare type of ovarian carcinoma. Description of these tumours have been rare and mostly covered in case reports. In some cases, molecular characterization by sequencing has been employed for guided therapy recommendations, however, functional chemosensitivity testing of targetable pathways using advanced in vitro cellular models such as organoids has not been reported so far. Here, we report on a case of ovarian cancer that was later identified as mesonephric-like adenocarcinoma at an advanced stage. MethodsThe tumour was characterized by molecular techniques including immunohistochemistry and whole-exome sequencing. At the same time, ovarian cancer organoids were established by adapting existing protocols for high-grade serous ovarian carcinoma. The organoids were subsequently used for functional in vitro chemosensitivity testing by treatment with standard-of-care chemotherapeutics cisplatin, paclitaxel, and the Poly (ADP-Ribose) Polymerase 1-inhibitor olaparib. Based on molecular characteristics, we also applied the inhibitor binimetinib, to target Mitogen-Activated Protein Kinase downstream of the KRAS Proto-Oncogene. Additionally, chemotoxicity testing with healthy fallopian tube organoids and high-grade ovarian cancer organoids was applied to determine the therapeutic window. ResultsImmunohistochemical analysis showed characteristic PAX8+, GATA3+, TFF1+, ER-, PR-, WT1- staining while the sequencing revealed mutations in 31 genes of which KRAS G12V and DYNC1H1 G4072S were annotated as (likely) pathogenic. The tumour was mismatch-repair proficient. Tumour-derived organoids proved to be highly resistant to standard-of-care chemotherapeutics cisplatin, paclitaxel, and olaparib, but sensitive to inhibition by binimetinib, which aligned well with the molecular characteristics. Direct comparison to healthy fallopian tube organoids and high-grade ovarian cancer organoids confirmed low cytotoxic potential underlining a feasible therapeutic window for binimetinib. ConclusionsFor the first time, we show that existing protocols for high-grade serous ovarian carcinoma can be used for the generation of organoids derived from mesonephric-like adenocarcinoma. These organoids could be used as an essential tool for functional precision medicine purposes. This functional data could be applied as an additional layer for molecular tumour boards diagnostics by supporting molecular datasets and even identify targetable pathways beyond genetic variations, thus offering novel therapeutic options particularly for rare and aggressive tumours.

Hormone replacement therapy (HRT) with oestrogen and progestogen is a common medical treatment for alleviating symptoms of menopause. Since 2015, its use has been increasing in the UK. Unscheduled bleeding can be a symptom of endometrial cancer, and guidelines state that women experiencing this should have an urgent referral for suspected endometrial cancer. However, unscheduled bleeding is also common in women taking HRT, particularly in the first few months after starting HRT or if there is a change in regimen. Current guidelines may result in women on HRT receiving referrals that are not necessary and undergoing unpleasant and invasive tests such as hysteroscopy. However, there is a lack of current information to guide recommendations. This protocol describes a cohort study in the ORCHID-e database of anonymised patient records from English primary care. We will use a cohort of women aged over 40 years starting on HRT with oestrogen and progestogen, age matched to women who have not started HRT. Exposure will be a prescription for oestrogen containing HRT with no previous prescription for oestrogen containing HRT in the previous year. Index date in each matched set will be the date of this prescription. Prescriptions for progestogen containing drugs will not be used to define the exposure, but this information will be extracted to describe the study population and for sensitivity analyses. Outcomes will be consultations for unscheduled bleeding, urgent referrals for suspected endometrial cancer, and diagnosis of endometrial cancer. Women will be followed up until they change exposure status or are otherwise censored. Women who start taking HRT in follow-up will re-enter the cohort in the exposed group. We will describe proportions of women with a code for consulting with unscheduled bleeding, proportions of those women referred for further investigation on the pathway for suspected endometrial cancer, and proportions diagnosed with endometrial cancer within one year of referral. We will investigate the diagnostic accuracy of unscheduled bleeding for endometrial cancer separately for women on HRT and those not on HRT. Analyses will be done by 6-month categories of time since index, age, calendar year, sociodemographic variables, risk factors for endometrial cancer, type of HRT.

BackgroundPlatelet count and C-reactive protein (CRP) are blood tests commonly used in primary care as part of diagnostic work up for symptomatic patients. Abnormal results of these tests can indicate an undetected cancer; however, it is not known whether the association between an abnormal test result and cancer risk varies by patient ethnicity. MethodsThis cohort study used routinely collected primary and secondary health care records in England with linkage to national cancer registry data. Included patients had a record of ethnicity, no prior malignancy, a platelet count or CRP record between 1st January 2010 and 31st December 2017, and were aged 40 years or over at the time of that test. Ethnicity was categorised as White, Asian, Black, Other, and Mixed. Multi-level logistic regression models estimated cancer incidence within one-year of testing, adjusted for age, sex, comorbidities, BMI, deprivation, and year of test. ResultsAmong 4,948,342 patients with a platelet record and 811,559 with a CRP record, one-year cancer incidence was highest among White patients and lowest among Asian patients. Following a normal platelet count, cancer incidence was 1.3% (95% CI 1.3-1.3%) for White patients and 0.63% (0.60-0.66%) for Asian patients; following thrombocytosis, incidence increased to 4.1% (4.0-4.2%) and 1.8% (1.5-2.0%), respectively. After a normal CRP result, cancer incidence was 1.5% (1.4-1.5%) for White patients and 0.79% (0.71-0.88%) for Asian patients, rising to 3.6% (3.5-3.7%) and 1.9% (1.7-2.2%) for a high CRP result, respectively. No significant interactions were found between ethnicity, blood test result, and overall cancer diagnosis, and similar diagnostic odds ratios (dOR) were observed across all ethnic groups. However, for colorectal cancer, Black patients with abnormal results showed higher diagnostic odds ratios (dOR) compared with White patients, relative to a normal result. The dOR for thrombocytosis was 11.1 (7.8-15.6) for Black patients versus 5.7 (5.4-6.0) for White patients (interaction p-value <0.001), and for raised CRP was 4.1 (2.6-6.6) for Black patients versus 2.5 (2.3-2.7) for White patients (interaction p-value=0.043). ConclusionThis large primary care study underscores the need for ethnically diverse cohorts when evaluating diagnostic tests to avoid widening healthcare inequalities.

BackgroundCystoscopy is a core component of haematuria investigations but is invasive and resource-intensive. GALEAS Bladder is a DNA-based diagnostic urine test that measures alterations in 23 bladder cancer-associated genes. ObjectiveTo assess the diagnostic performance and clinical utility of GALEAS Bladder as a molecular triage tool in real-world haematuria investigation pathways. MethodsPatients referred for urgent investigation of haematuria were prospectively enrolled across seven UK NHS Urology Departments between October 2024 and June 2025. Urine samples were collected prior to cystoscopy and analysed using the GALEAS Bladder assay (Nonacus Clinical Services, UK). Assay results were compared with cystoscopy findings. Key Findings and LimitationsCystoscopic findings and GALEAS Bladder results were available for 964 participants, including 77 (8.0%) newly-diagnosed with pathology-confirmed BC. The assay demonstrated an overall sensitivity of 92.2% (95% CI: 84.0-96.4%), specificity of 92.0% (95% CI: 90.0-93.6%), and negative predictive value (NPV) of 99.3% (95% CI: 98.4-99.7%) for the diagnosis of BC. For the diagnosis of high-grade BCs, sensitivity was 97.2% (95% CI: 85.8-99.5%) with an NPV of 99.9% (95% CI:99.3-100.0%). Limitations include an absence of subsequent diagnoses for participants with positive GALEAS Bladder test results in the absence of cystoscopically-visible tumour. Conclusions and Clinical ImplicationsGALEAS Bladder is a clinically implementable molecular urine test with very high sensitivity and specificity for the diagnosis of new cases of BC in patients undergoing urgent investigation of haematuria, especially for high-grade BCs. Clinical adoption could permit the molecular triage of haematuria patients to immediate or deferred cystoscopy.

BackgroundSerous-tubal intraepithelial-carcinoma (STIC) is considered the principal precursor of tubo-ovarian high-grade serous-carcinoma (HGSC), yet its biological uniformity, progression risk and potential response to poly(ADP-ribose) polymerase-inhibitors (PARPi) remain poorly defined. MethodsWe performed trajectory-based transcriptomic reconstruction of Fallopian-tube epithelial regions spanning normal epithelium, precursor lesions, STIC, and invasive carcinoma using pseudotime inference and publicly available spatial transcriptomic data. Gene expression and pathway dynamics were defined along pseudotime, and interpatient heterogeneity examined at both lesion and patient levels. Transcriptional signatures associated with PARPi-resistance were quantified across STICs, BRCA-mutant and BRCA-wild type subtypes. ResultsTrajectory inference captured a continuous transcriptional progression from normal epithelium to HGSC, with STICs occupying heterogeneous evolutionary positions rather than a single precursor state. Incidental isolated STICs(STICi) spanned early to later pseudotime states and frequently aligned with loss-of-cilium organisation and less advanced epithelial phenotypes. STICs associated with concurrent cancer(STICc) exhibited more advanced malignant progression signatures, including cell-cycle activation, epithelial-to-mesenchymal transition (EMT), interferon signaling, and DNA-repair. Histologically similar lesions occupied divergent pseudotime positions with marked interpatient heterogeneity. PARPi resistance-associated signatures were variably enriched across precursor and precancer-stage lesions, with persistence into invasive disease. ConclusionsSTICs are heterogeneous and occupy distinct evolutionary positions along a continuum, highlighting potentially different progression risks from normal epithelium to HGSC. STICi differ from STICc which harbour signatures of more advanced malignant progression. Heterogeneity in PARPi resistance-associated programs in STICs cautions against uniform (non-stratified) use of PARPi-based primary prevention strategies. Future research should explore evolutionary-trajectory informed biomarkers for risk stratification and early interception strategies. Translational relevanceSerous tubal intraepithelial-carcinoma (STIC) represents a precursor of tubo-ovarian high-grade serous carcinoma (HGSC), offering unique translational opportunities for early detection, risk stratification, and prevention. However, its phenotypic uniformity, progression risk and therapeutic drug response are not fully understood. Our study shows that STICs are not a uniform precursor state, but instead occupy a range of evolutionary positions along a continuum from normal epithelium to invasive HGSC. This heterogeneity reflects continuous transcriptional variation rather than discrete precursor categories. We report gene expression dynamics and molecular signature changes across the malignant transformation timeline. We further illustrate that precursor lesions exhibit phenotypic variability that impacts therapeutic drug resistance-associated programs. Our data highlight potential disease biomarkers defined by evolutionary trajectory inference and transcriptional processes associated with drug resistance operating in precursor lesions. These findings may help improve our ability to distinguish clinically significant lesions and inform targeted interception strategies.

ObjectivePopulation-based information regarding adherence to first-line chemotherapy in epithelial ovarian cancer is scarce. This study aimed to evaluate chemotherapy adherence, reasons for chemotherapy modifications, and associations with overall survival. MethodsAdvanced-stage epithelial ovarian cancer patients diagnosed between January 2015 and December 2021 were identified from the Netherlands Cancer Registry. Patients who underwent cytoreductive surgery combined with platinum- and taxane-based chemotherapy were included. Patients were categorized into two groups: adherent (patients without modifications) and non-adherent (patients with modifications: dose reduction, chemotherapy interruption, and/or reduction in chemotherapy cycles). Reasons for modifications were assessed. Kaplan-Meier survival curves and Cox proportional hazards models were used to analyze overall survival. ResultsAmong the cohort (N = 3,687), 54% of patients underwent chemotherapy modifications. Dose reduction (38%) was the most common, followed by interruption (24%) and reduction in chemotherapy cycles (9%). Non-adherence was associated with poorer performance scores, higher comorbidity indices, and undergoing primary cytoreductive surgery. Neurotoxicity and hematologic toxicity were the primary reasons for modifications in platinum (33% and 37%) and taxane (47% and 35%) agents. No association with survival was found for dose reduction and interruption. However, reduction in chemotherapy cycles was associated with lower 5-year overall survival (32% (95% CI 26%-38%) vs. 36% (95% CI 34%-38%)), remaining significant after multivariable adjustment (hazard ratio 1.36; 95% CI 1.17-1.59). ConclusionA significant proportion of Dutch advanced-stage epithelial ovarian cancer patients undergo chemotherapy modifications. No impact on overall survival was found for dose reduction or chemotherapy interruption, warranting prospective studies. Reduction in chemotherapy cycles was negatively associated with overall survival, possibly reflecting underlying treatment ineffectiveness. Key messagesO_ST_ABSWhat is already known on this topicC_ST_ABSGuideline-recommended chemotherapy for advanced epithelial ovarian cancer is often difficult to deliver in routine practice, and real-world data on adherence and its impact on survival are limited. What this study addsIn this nationwide retrospective cohort, over half of patients experienced chemotherapy modifications; dose reductions and interruptions were not associated with poorer overall survival, whereas a reduction in the number of cycles showed an association with worse outcomes, although this may partly reflect underlying disease severity or treatment response. How this study might affect research, practice or policyOur findings suggest that standard dosing and treatment duration of six cycles may not always be necessary, emphasizing the need to tailor treatment plans to optimize both efficacy and tolerability in advanced-stage epithelial ovarian cancer patients

BackgroundOvarian cancer (OC) progression and chemoresistance are closely linked to dysregulated oncogenic signaling, including Wnt/{beta}-catenin pathways that contribute to cancer stem-like traits. However, the upstream mechanisms connecting cytoskeletal regulation to Wn/{beta}-catenin signaling in OC remain incompletely understood. Keratin 17 (KRT17), a type I intermediate filament protein, has been implicated in tumor progression, but its mechanistic role in OC requires clarification. MethodsGene Expression Omnibus (GEO) datasets and clinical specimens were analyzed to assess KRT17 and EPN1 expression and prognostic significance. Functional assays, xenograft models, co-immunoprecipitation, ubiquitination analyses, and rescue experiments with wild-type and ubiquitination-resistant EPN1 mutants were performed to investigate molecular mechanisms. ResultsKRT17 expression was elevated in OC tissues and correlated with poor patient survival. KRT17 depletion suppressed proliferation, migration, stem-like properties, tumor growth, and cisplatin resistance. Mechanistically, KRT17 interacted with EPN1 and weakened its association with the E3 ligase SMURF1, reducing SMURF1-mediated ubiquitination at lysine 107 and preventing proteasomal degradation. Stabilized EPN1 was associated with increased {beta}-catenin abundance and stemness-associated markers, and enhanced self-renewal capacity. ConclusionsThese findings identify a KRT17-EPN1 axis that links intermediate filament dynamics to ubiquitin-dependent regulation of EPN1 stability and Wnt/{beta}-catenin signaling outputs in ovarian cancer.

ObjectivesCervical cancer is a preventable disease, and a properly implemented screening programme can reduce its incidence and mortality and potentially save resources. This study aimed to evaluate the cost-effectiveness of options for potential transformation of the nationwide screening programme in the Czech Republic, especially considering recent changes in HPV DNA testing recommendations. MethodsA microsimulation model was developed to assess the cost-effectiveness and health benefits of alternative screening strategies in the Czech Republic. The model simulated annual life cycles of women from age 15, comparing combinations of cytology and HPV testing. Input parameters used were obtained from national registries in the Czech Republic and from published literature. The analysis was conducted from the perspective of healthcare payers. Costs (2025 EUR) and LYs were discounted at a rate of 3% annually. Probabilistic sensitivity analysis was conducted. ResultsThe CEA showed that, compared to the current setting (annual cytology with co-test at 35, 45, 55), only specific co-testing strategies lead to a decrease in incidence and mortality but differ in benefits and economic efficiency. The lowest ICER was reported for a strategy combining cytology at two-year intervals and co-testing at four-year intervals from ages 30 to 65. Sensitivity analysis showed that the current strategy has the highest probability of cost-effectiveness at {euro}31,000 per LY gained. At higher values, this is replaced by a strategy with a 3-year interval co-test. ConclusionsBased on the models presented, co-testing appears to be cost-effective. The actual willingness to pay threshold will facilitate selection of the most-appropriate strategy.

BackgroundEpidermal Growth Factor Receptor (EGFR) inhibitors, while effective in oncology, are associated with under-characterized ocular adverse events (AEs). Prior studies have been limited in scope, lacking a comprehensive, class-wide analysis of the full spectrum of ocular toxicity, particularly for newer agents. MethodsWe conducted a disproportionality analysis of the FDA Adverse Event Reporting System (FAERS) (2001-2025). Twelve EGFR-targeted agents were evaluated against a pre-specified set of ocular MedDRA Preferred Terms. To ensure robust signal detection, a significant association was defined by [&ge;]3 co-reported cases, a Proportional Reporting Ratio (PRR) [&ge;]2.0, and a false-discovery-rate adjusted p-value <0.05. ResultsAmong 6,976,462 drug-event combinations, 20 met all signal criteria for Eyelash Abnormalities, Ocular Surface Disease, or Vision-Threatening and Intraocular Events. Trichomegaly demonstrated extreme disproportionality (e.g., panitumumab PRR= 465.3, 95% Confidence Interval [CI], 247.7-874.3). A consistent pattern of ocular surface toxicity (conjunctivitis, keratitis, blepharitis) was observed across multiple tyrosine kinase inhibitors and monoclonal antibodies, indicating a class-wide effect. Signals for serious events included corneal perforation (erlotinib, n= 7, PRR=13.9, 95% CI= 6.6-29.4) and optic neuropathy (erlotinib, n= 6, PRR= 2.9, 95% CI= 1.3-6.4). ConclusionThis analysis confirms a strong, class-wide signal for ocular toxicity across the spectrum of EGFR inhibitors, from characteristic eyelid changes to sight-threatening complications. These findings underscore the necessity for proactive ophthalmologic monitoring, including baseline assessment, in patients receiving these therapies to preserve vision and maintain quality of life during cancer treatment.

PurposeCastration-resistant prostate cancer (CRPC) is characterized by marked clinical heterogeneity and poor long-term survival, underscoring the need for tools that can rapidly and reliably individualize patient risk. While several prognostic models exist, their complexity has limited routine clinical use. Here, we developed and validated PROGRESS (PROstate cancer Global Risk Evaluation and Stratification Score), a simplified prognostic score, derived through machine learning-guided feature selection, to enhance risk stratification and support individualized, risk-informed clinical decision-making. MethodsPROGRESS was developed using baseline data from 2,035 metastatic CRPC patients enrolled in four different phase III trials. An unsupervised machine-learning approach was applied to identify latent patient subgroups with distinct survival outcomes irrespectively of allocated treatment arm, followed by classical multivariable modelling to derive a simple and straight-forward prognostic score based on routinely available objective laboratory variables. External validation was performed in three independent datasets comprising metastatic CRPC patients treated across different therapeutic settings (n=1,239) and non-metastatic CRPC patients managed with standard care (n=660). Overall survival was assessed using Kaplan-Meier and Cox regression analyses. ResultsUnsupervised modelling identified two patient risk subpopulations with significantly different overall survival rates (median 27.4 vs 17.7 months; hazard ratio [HR] 2.20, 95% CI 1.91-2.54; p<.00001). Feature contribution analysis yielded three independent predictors -PSA, ALP, and AST-used to build PROGRESS. In the training cohort, PROGRESS demonstrated strong discrimination (AUC 0.89). Using a prespecified cut-off, patients classified as increased risk had significantly shorter survival than low-risk patients (median 18.3 vs 25.6 months; HR 1.72, 95% CI 1.50-1.97; p<.0001). PROGRESS prognostic performance was consistent across all validation cohorts, including metastatic and non-metastatic disease, with HRs ranging from 1.74 to 3.46 (all p<.0001). ConclusionsBy integrating machine-learning-based pattern discovery with classical statistical modelling, PROGRESS provides a simple, objective, and clinically accessible approach for individual risk stratification in CRPC. Its reliance on three inexpensive, routinely measured laboratory parameters would facilitate practical implementation in clinical settings, enhancing visibility of underlying disease aggressiveness for individual clinical decision-making. PROGRESS could represent a pragmatic first step toward improving patient selection for clinical trials while identifying regulatory meaningful endpoints achievable in a sizeable patient population; further validation in prospective clinical studies and real-world datasets would allow to confirm its clinical utility and generalizability. PROGRESS can be freely accessed for research use only at the following link: https://dev.ai.topazium.com.

BackgroundLynch syndrome (LS) is a cancer susceptibility syndrome caused by germline pathogenic variants in DNA mismatch repair (MMR) genes. Due to increased risk of colorectal cancer (CRC), enhanced colonoscopic surveillance is recommended for heterozygote MMR-carriers. ObjectiveUsing a registry of English LS patients linked to digital National Health Service records, we aimed to assess adherence of MMR-carriers to national surveillance guidelines, and to determine the impact of surveillance on CRC incidence and mortality. DesignWe described the frequency of colonoscopies in 4,732 MMR-carriers and used logistic regression to determine predictors of surveillance adherence. For MMR-carriers with a record of surveillance and those without, we: estimated age-specific annual CRC incidence rates (AS-AIRs) and cumulative lifetime risks, assessed for stage-shift by comparing CRC stage distributions and stage-specific AS-AIRs, and estimated risks of death from CRC and any cause using Kaplan-Meier methods and Cox Proportional Hazards regression. ResultsSurveillance at a mean interval of [&le;] 3 years (n=3028) was associated with a decrease in CRC-specific and all-cause mortality, without an associated change in total CRC incidence, even after multivariate adjustment. No strong evidence of stage-shift was observed. Colonoscopic surveillance at a mean interval of [&le;] 2 years (n=1569) was associated with an increase in total CRC incidence. Incidence of early-stage cancers was also higher, with no corresponding decrease in late-stage cancers, which may reflect the short follow-up period or the impact of overdiagnosis. ConclusionThe observed reduction in all-cause mortality amongst regularly-surveilled MMR-carriers may indicate an impact of surveillance on CRC-specific mortality, though in the context of a non-randomised study likely reflects the influence of selection bias. KEY MESSAGES OF ARTICLEO_ST_ABSWhat is already known on this topicC_ST_ABSRegular surveillance colonoscopy is recommended in Lynch syndrome, though evidence to support this remains mixed. We searched PubMed for articles published from inception to 01/05/2024 using the terms "Lynch syndrome", "HNPCC", "colonoscopy", "sigmoidoscopy", "surveillance", and "screening". We found one controlled trial and several small analytical studies dating from the early 2000s which compared surveilled and non-surveilled populations and found surveillance to be associated with reduced colorectal cancer (CRC) incidence and improved survival. More recent longitudinal observational studies, most without comparator groups, found a high incidence of CRC in LS populations despite being resident in countries where surveillance was recommended. A small number of studies directly assessed time since last colonoscopy against CRC incidence and stage with mixed findings. Finally, cross-sectional comparisons between countries of CRC incidence rates and surveillance interval recommendations found no relationship between the two1,2. What this study addsHere, we conduct an observational cohort study on a large national cohort of MMR germline pathogenic variant (GPV) carriers (MMR-carriers) in England (n=4,732), comparing CRC incidence and mortality in individuals with a record of regular surveillance to those without. Through linkage of the English National Lynch Syndrome Registry to Hospital Episodes Statistics data, we are uniquely able to study a comprehensive national population of MMR-carriers and identify the dates on which colonoscopies were undertaken over time, allowing assessment of adherence to national surveillance guidelines and the impact this has on CRC outcomes. Notably, receipt of regular colonoscopy was strongly associated with deprivation as well as ethnicity. The results show that regular surveillance at an average interval of 3 years (or less) is not associated with a reduction in CRC incidence when compared to less frequent surveillance, but an apparent decrease in both CRC-specific and overall mortality is observed, even after adjustment for confounding variables. Conversely, regular surveillance at an average interval of 2 years (or less) is associated with an increase in CRC incidence when compared to less frequent surveillance, which may suggest increased diagnosis of early-stage cancers or, due to the absence of a reduction in late-stage cancers, overdiagnosis. The observed impact of surveillance on overall mortality may demonstrate the impact of surveillance on CRC-specific mortality, or, in the context of an observational (non-randomised) study, indicate that the results are subject to selection bias. How this study might affect research, practice, or policyEvidence for the benefit of surveillance colonoscopy remains mixed. Whilst polypectomy would be anticipated to prevent CRC development (thus reducing CRC incidence), several studies have observed increased frequency of CRCs in MMR-carriers undergoing frequent surveillance colonoscopy, which may reflect overdiagnosis. The selection bias inherent to observational studies of surveillance renders mortality outcomes challenging to interpret. Randomised controlled trials of colonoscopic surveillance in MMR-carriers are required for effectiveness of this intervention to be accurately assessed. Given ethical and feasibility challenges, randomised controlled trials might be complemented by quasi-experimental designs using advanced observational methods for assessing effectiveness.

BackgroundChemo-resistance remains a major clinical challenge in Oral Squamous Cell Carcinoma (OSCC), attributed to the intrinsically resistant cells. Although tumour-derived extracellular vesicles (EVs) have been implicated in cell-cell communication, their role in propagating chemo-resistance remains poorly defined. This study aims to identify salivary EV-associated miRNAs capable of predicting chemoresistance and to delineate the role of miR-1307-5p in modulating CSC-driven therapeutic refractoriness. MethodsSalivary EV-derived expression profile of miR-1307-5p was assessed by qPCR in chemo resistant OSCC patients and further validated in TCGA small RNA sequencing datasets. Expression was validated by qPCR and correlated with clinicopathological outcomes. Functional assays including cell-cycle analysis, apoptosis, migration/invasion, 3D spheroids, angiogenesis, and CAM assays were performed in miR-1307-5p inhibited CD44 CSC subpopulation compared to its vehicular control. Transcriptomic profiling cross-referencing with TCGA was conducted to identify potential novel targets of miR-1307-5p. Chemo-sensitisation was assessed by treating the knockdown chemo resistant cells with low dose cisplatin and validating it using in-vitro functional assays and orthotopic xenograft model. ResultsmiR-1307-5p was significantly elevated in salivary EVs of chemo resistant OSCC patients and correlated with poor overall survival (p = 0.03). The miRNA was markedly enriched in endogenously resistant CD44 CSCs. Silencing of miR-1307-5p induced G2/M arrest, triggered apoptosis, impaired invasion, and reduced angiogenesis both in-vitro and in ex-vivo assays. Transcriptomic profiling, TCGA validation, and integrative pathway analysis identified key oncogenic hubs which converge on PI3K-AKT, MAPK/ERK, and YAP signalling pathways governing EMT. Inhibition of miR-1307-5p restored cisplatin sensitivity in resistant CSCs, with low-dose cisplatin producing substantial tumour suppression in-vitro and in-vivo. Reduced CD44 expression in xenograft models confirmed CSC reprogramming. EVs from anti-miR-treated cells confer chemo sensitisation upon uptake by resistant CSCs. Xenograft models substantiated that EVs can initiate tumour formation and that EV-mediated delivery of anti-miR-1307-5p drives significant tumour regression. ConclusionThis study identifies salivary EV-derived miR-1307-5p as a clinically relevant biomarker of chemoresistance in OSCC and reveals its mechanistic role in sustaining CSC-driven therapeutic failure. Targeting miR-1307-5p offers a promising avenue for restoring cisplatin sensitivity and developing exosome-based therapeutic strategies. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=150 SRC="FIGDIR/small/709730v1_ufig1.gif" ALT="Figure 1"> View larger version (38K): org.highwire.dtl.DTLVardef@19f88e0org.highwire.dtl.DTLVardef@d36b95org.highwire.dtl.DTLVardef@3c2579org.highwire.dtl.DTLVardef@c04ef5_HPS_FORMAT_FIGEXP M_FIG C_FIG

Background and ObjectiveDespite its prevalence, bladder cancer (BC) remains an unsolved pathogenesis. It is believed that TRPC6 channels have unique electrophysiological properties that contribute to intracellular Ca2+ signalling and tumorigenesis in cells. However, the mechanism by which TRPC6 contributes to BC progression and intracellular Ca2+ homeostasis remains unclear. MethodIn this study, TRPC6 expressions in paired BC and adjacent normal tissues were measured by immunohistochemistry. A KEGG pathway enrichment analysis was conducted to determine TRPC6s potential contribution to BC. Ca2+ imaging analysis was performed to explore the contribution of TRPC6 in the BC cell. Flow cytometry and Cell Counting Kit-8 assay were performed to explore the effects of TRPC6 on the proliferation of BC. The impacts of TRPC6 SOCE on PI3K/Akt/mTOR pathway were measured by western blot. Based on the above bunch of studies, TRPC6 was found to be overexpressed in human BC tissue, which correlated with poor survival rates for patient overall survival (OS). We used the TRPC6-specific antagonist SAR7334 to explore and reveal significant inhibition of BC cell proliferation. Mechanistically, TRPC6 mediated cytosolic Ca2+ and regulation of SOCE, leading to the activation of the IP3K/AKT/mTOR pathway. ResultsWe found that SAR7334 arrested the phosphorylation of PI3K and Akt, thus causing a significant decrease in phosphorylated mTOR. Similar effects were observed for the SOCE-specific antagonist MRS1845. In contrast, the Akt inhibitor MK2206 did not alter the SOCE in BC cells. Conclusionsour results indicate that the pharmacological inhibition of TRPC6 arrests tumour cell proliferation through SOCE targeting the PI3K/Akt/mTOR pathway. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=101 SRC="FIGDIR/small/702689v1_ufig1.gif" ALT="Figure 1"> View larger version (28K): org.highwire.dtl.DTLVardef@bf783aorg.highwire.dtl.DTLVardef@6e3487org.highwire.dtl.DTLVardef@13aad23org.highwire.dtl.DTLVardef@13cfe7e_HPS_FORMAT_FIGEXP M_FIG C_FIG Schematic of proposed pathway. A model for summarizing TRPC6 contributes to cell proliferation in bladder cancer. TRPC6 specific inhibitor SAR7334 alter cellular cytosolic Ca2+ and SOCE, thus arrested phosphorylation of the PI3K/Akt pathway and the proliferation of bladder cancer cells.

PurposeProgastrin, aberrantly expressed in colorectal cancer (CRC), is an established trophic factor for tumour epithelial cells. Whether it also promotes CRC progression by reprogramming stromal fibroblasts remains unclear. We investigated progastrin-induced colonic fibroblast activation and its functional consequences on CRC cell migration. MethodsFibroblast activation was assessed in the colonic mucosa of hGAS mice and in the human normal colonic fibroblast line CCD18Co exposed to synthetic progastrin. The impact of tumour-derived progastrin on epithelial cell motility was analysed using HCT116 cells expressing a control shRNA (shLuc) or a progastrin-targeting shRNA (shPG) in transwell migration assays performed with or without fibroblasts. Candidate paracrine mediators were evaluated by RT-qPCR, ELISA and neutralization experiments, and signalling was interrogated using the PI3K inhibitor LY294002. ResultsColonic fibroblasts from hGAS mice displayed stromal FAP and SMA expression, indicating fibroblast activation in vivo. In CCD18Co cells, progastrin increased FAP and SMA protein levels. Fibroblasts enhanced HCT116 cell migration. This effect was stronger when tumour cells expressed progastrin or when fibroblasts were preconditioned by progastrin-producing HCT116 cells. Progastrin induced CXCL12/SDF-1 and CXCL8/IL-8 expression and secretion by fibroblasts, and neutralization of either chemokine abrogated the additional migratory effect conferred by progastrin-activated fibroblasts. Progastrin triggered sustained Akt phosphorylation in fibroblasts, while PI3K inhibition suppressed CXCL12 and CXCL8 secretion and abolished fibroblast-dependent tumour cell migration. ConclusionThese data identify a stromal dimension of progastrin signalling in CRC and support a model in which tumour-derived progastrin activates colonic fibroblasts and elicits a PI3K/Akt-dependent paracrine programme that enhances CRC cell migration.

BackgroundMulticancer early detection tests could be used for cancer screening, but may lead to harms, including false positive results and overdiagnosis of indolent tumours that would not have become clinically evident during that persons lifetime. We assessed the potential for these screening harms in the context of future population-based screening with a multicancer early detection test. MethodsWe used a microsimulation model to assess potential population-level impacts of screening at ages 50-75 years with a multicancer early detection test in Canada. We assumed high test specificity (97-99.1%) and test sensitivity increasing with cancer stage. The model includes latent indolent cancers that would not be diagnosed within that persons lifetime but can be overdiagnosed through screen-detection. We calculated the yearly and cumulative lifetime probabilities of screening overdiagnosis and false positive test results, assuming a range of preclinical screen-detectable periods (2-5 years). ResultsAn estimated 2.1-6.0% of all yearly screen-detected cancers with a multicancer screening test were predicted to be overdiagnoses across scenarios. The proportion of overdiagnosis varied by site, and strongly increased with age, going from 1% at age 50 to over 10% of screen-detected cancers by age 75. The test positive predictive value ranged from 15.9%-77.6%, meaning that there could be 0.3-5.3 false positives with no underlying cancer for every true cancer case detected by the test. ConclusionPopulation-level multicancer screening with a multicancer early detection test would likely not lead to substantial screen-related overdiagnosis. Healthcare systems should consider how screening false positives may increase their diagnostic service caseload.

ObjectiveThe purpose of the present study is to describe the survival outcomes of patients with low-grade serous ovarian cancer (LGSOC) in the post-operative setting from a tertiary gynecologic oncology referral centre in Quebec, including evaluation of patient characteristics, clinical outcomes and prognostic factors. MethodsThe study included 25 patients: 1) with a post-surgical histopathologic diagnosis of a low-grade serous tumour of the ovary, 2) underwent primary cytoreductive surgery prior to adjuvant therapy, and 3) for whom clinical data was available. Clinical and demographic features were characterized by descriptive statistics. Clinical endpoints of progression-free survival (PFS) and overall survival (OS) were assessed, utilizing the Kaplan-Meier method for estimating survival probabilities. ResultsThe median age of this cohort was 61 years (range, 26-81). Median OS was 140.6 months in patients with no residual disease (R0), 71 months in patients with microscopic residual disease (R1), and 27.7 months in patients with macroscopic residual disease (R2) (p=.001). Residual disease was also found to significantly impact PFS (p=.008). Administration of adjuvant chemotherapy failed to improve survival outcomes altogether (PFS, p = .270; OS, p = .300). ConclusionsThis study supports the shifting consensus that optimal cytoreductive surgery, where feasible, is paramount for successful treatment of LGSOC. Furthermore, treatment with adjuvant chemotherapy may lead to worse survival outcomes.

Purpose: High-quality population-based cancer registration data is crucial for cancer research. We describe the methodology that the National Disease Registration Service (NDRS) established to accelerate provision of cancer registration data in England to support timely analysis of the NHS-Galleri trial (NCT05611632), assessing the clinical utility of a multi-cancer early detection test for population screening. Methods: NDRS established two accelerated cancer registration products for trial participants: Expedited Core (providing complete stage data 6 months after diagnosis) and Expedited Comprehensive (providing [&ge;]6 months of follow-up clinical pathway data 11 months after diagnosis). NDRS used pre-existing data sources and methodology, plus enhanced dedicated data liaison support. Timeliness and concordance of accelerated data was assessed 6, 11, and [&ge;]19 months after diagnosis. Results: At 6-10 months after diagnosis, most (98.0%) registrations achieved Expedited Core status; at >10 months, all (100%) registrations were Expedited Comprehensive. For fact of malignant cancer and stage III/IV diagnoses, concordance between registrations at 6 and [&ge;]19 months was 96.7% and 95.6%, respectively. Conclusions: NDRS delivered staged cancer registration data 6 months after diagnosis. High concordance at 6, 11, and 19 months provides confidence in the accelerated data. This supports reporting NHS-Galleri findings more promptly than possible via routine data.

Although the retinoblastoma protein (pRB) is functionally inactivated by hyperphosphorylation in the majority of colorectal cancers (CRC) - with RB1 rarely mutated and even amplified at the genomic level - three critical gaps remain unaddressed: no study has systematically compared which first-line chemotherapeutic agent best synergizes with CDK4/6 inhibition using head-to-head quantitative analysis; functional differences between palbociclib and abemaciclib in chemotherapy combinations have not been characterized in CRC; and direct genetic evidence of RB dependency in this combinatorial context is lacking. Here, we addressed these gaps by evaluating palbociclib and abemaciclib combined with oxaliplatin, 5-fluorouracil, and SN-38 in HCT116 CRC cells, with validation in SW480 cells, RB1-silenced HCT116 cells (shRNA-RB), and non-tumoral intestinal epithelial cells (IEC-6), using quantitative drug interaction analysis (Chou-Talalay), cell cycle profiling, apoptosis assessment, and pRB phosphorylation measurement. Oxaliplatin was the most consistently synergistic partner for both CDK4/6 inhibitors (CI < 1 across all tested concentrations), while combinations with SN-38 yielded variable results and 5-FU combinations approached additivity. The oxaliplatin combination reinforced G1 arrest and enhanced cell death, with abemaciclib producing more pronounced apoptotic induction than palbociclib - an effect not explained by differential pRB target engagement (both inhibitors reduced pRB Ser807/811 phosphorylation by [~]50%), likely reflecting abemaciclibs broader kinase inhibitory profile. shRNA-mediated RB1 silencing partially attenuated the combinatorial effect, providing direct genetic evidence that the synergy is RB-dependent. Importantly, the combination did not significantly potentiate oxaliplatin cytotoxicity in non-tumoral IEC-6 intestinal epithelial cells, in contrast to the pronounced enhancement observed in tumor cells, and synergistic benefit was preserved at sub-cytotoxic inhibitor concentrations. These findings identify oxaliplatin as the optimal chemotherapeutic partner for CDK4/6 inhibition in CRC, with a mechanism involving RB-dependent potentiation of apoptosis that is preferentially active against tumor cells and maintained at clinically relevant inhibitor doses.

IntroductionBRCA1/2 alterations are increasingly recognized as biologically and clinically relevant features in prostate cancer, yet the prognostic and therapeutic significance of zygosity status remains uncertain. Understanding differences between monoallelic and biallelic inactivation may refine risk stratification and guide therapeutic decision-making. Materials and MethodsA retrospective, desk-based observational analysis was performed using publicly accessible datasets from TCGA-PRAD (primary disease) and SU2C/PCF (metastatic disease). BRCA1/2 status was categorized as wild-type, monoallelic, or biallelic based on mutation, copy-number, and loss-of-heterozygosity profiles. Overall survival was evaluated using Kaplan-Meier estimates and Cox models. Systemic therapy outcomes were assessed by treatment class, incorporating exploratory interaction tests. ResultsIn TCGA-PRAD (n=300), OS did not significantly differ by zygosity (global log-rank p=0.45), with median OS of 80.0 months (wild-type), 78.0 months (monoallelic), and 55.0 months (biallelic). In SU2C/PCF (n=200), zygosity stratified outcomes significantly (global log-rank p=0.04): median OS was 22.0 months (wild-type), 14.0 months (monoallelic), and 16.0 months (biallelic). Treatment analyses showed ARSI exposure improved OS in wild-type disease (HR 0.60; 95% CI 0.38-0.95), while interaction testing suggested potential heterogeneity without statistical confirmation (interaction p=0.092). PARP inhibitor exposure showed directionally favorable HRs in wild-type and monoallelic groups but no significant interaction (interaction p=0.757). No therapy class demonstrated consistent effect modification by zygosity. ConclusionBRCA1/2 zygosity shows prognostic relevance in metastatic prostate cancer but not clearly in primary disease. While zygosity did not consistently modify systemic therapy associations in this dataset, findings support zygosity-aware reporting as a practical tool for molecular stratification and future research design.

PurposePrognosis in metastatic non-seminomatous germ cell tumors (mNSGCT) is currently guided by the IGCCCG classification, which incorporates tumor markers, organs involved with metastatic disease, and primary site but not histologic subtype. We aimed to evaluate whether specific histological components provide additional prognostic information in a large international mNSGCT cohort. Patient and MethodsWe analyzed clinical, pathologic, and outcome data from 662 patients with mNSGCT across multiple international centers. Cox regression and multivariable stepwise models were used to evaluate the impact of age, tumor histology, serum markers, primary site of disease, chemotherapy, IGCCCG, and post-chemotherapy surgery on overall survival. Analyses were performed using both complete-case and imputed datasets to account for missing values. ResultsThe presence of any percentage of embryonal carcinoma (EC) was independently associated with improved overall survival HR 0.603 (95% CI: 0.37-0.98, p=0.040), whereas yolk sac tumor (YST) predicted worse prognosis in complete-case analysis HR 2.27 (95% CI: 1.43 - 3.61 p = 0.001). Choriocarcinoma was also associated with a HR 1.58 (95% CI: 1.08 - 2.32 p= 0.019) adverse outcomes. IGCCCG risk classification remained a strong predictor of mortality HR up to 8.9 for Poor vs Good risk, (95% CI: 4.63 - 17.09 p < 0.001), but histologic components added significant independent prognostic value. Post-chemotherapy retroperitoneal lymph node dissection (RPLND) conferred a substantial survival benefit HR 0.44 (95% CI: 0.258 - 0.754 p=0.003). Interestingly, teratoma was not associated with mortality but was linked to younger age, testicular primaries, and higher likelihood of residual disease requiring surgery. ConclusionsHistological composition, particularly the presence of EC or YST, has a significant and independent impact on survival in mNSGCT, beyond established risk classifications. Integration of histological subtypes may enhance prognostic accuracy and guide individualized treatment strategies in advanced germ cell tumors.