Frontiers in Oncology

Objective -- To analyze the incidence dynamics of uveal melanoma (UM) in Irkutsk Oblast over the period 2023 -- 2025, assess the structure of newly diagnosed cases by territory and stage, and compare the outcomes of anticancer care with those of other constituent entities of the Russian Federation that lead in registered prevalence of malignant neoplasms (MN) of the eye and adnexa. Materials and Methods. Data from the cancer registry of the Irkutsk Regional Oncology Dispensary (IROD) for 2023 -- 2025 and information from the state reference book "The State of Cancer Care for the Population of Russia in 2025" (P.A. Herzen Moscow Oncology Research Institute) were used. The analysis included cases of MN of the eye and adnexa (ICD - 10 code C69) of uveal localization. Incidence rates per 100,000 population, stage distribution, proportion of morphological verification, mortality, and the cohort of patients under follow-up were calculated. Results. Over three years, 112 new cases of UM were registered in Irkutsk Oblast (37, 36, and 39, respectively), corresponding to a crude incidence rate of approximately 1.6 per 100,000 population per year. In 2025, the region ranked 3rd in the Russian Federation for the number of newly registered patients with MN of the eye (45 individuals) and 1st for the registered prevalence rate (15.6 per 100,000). The highest proportion of stage I disease (58.3 %) and one of the lowest mortality rates (1.9 %) were observed. Conclusions. Irkutsk Oblast is characterized by a consistently high incidence of UM, accompanied by a favorable stage distribution and low mortality, reflecting both the biological and geographic features of the region and an adequate level of organization of ophthalmic oncology care.

Background: Human papillomavirus (HPV)-positive oropharyngeal squamous cell carcinoma (OPSCC) shows higher prevalence of lymph node metastases than HPV-negative OPSCC. It remains unclear whether HPV-positive and HPV-negative OPSCC differ in occult metastasis risk, which is relevant to personalize elective nodal treatment volumes. Methods: We performed a retrospective analysis of 780 untreated OPSCC patients from four European centers. Patterns of lymph node level involvement (LNLs I, II, III, IV, V, VII) were assessed using a consensus diagnosis integrating imaging (CT, MRI, PET) and pathological findings from surgically treated patients. Occult metastases were assessed in 288 surgical patients with paired clinical and pathological information. Results: HPV-positive patients more frequently presented with ipsilateral LNL II involvement, particularly in early T-stage disease (80.6% [75.6-84.8] vs. 54.8% [47.2-62.2]). However, the probability of downstream involvement in LNLs III and IV conditioned on upstream LNL involvement was similar between HPV-positive and HPV-negative patients. Similarly, contralateral involvement conditioned on midline extension of the primary tumor was similar. The overall proportion of occult metastases, defined as clinically false-negative LNLs divided by pathologically positive LNLs, was lower in HPV-positive patients (24.7% [19.7-30.1] vs. 48.7% [39.7-57.8]). Conclusions: HPV-positive OPSCC showed higher prevalence of LNL involvement, especially in ipsilateral LNL II and early T-stage tumors. The data to directly compare occult metastasis risk has important limitations and does not allow definitive conclusions. However, the presented data suggests that higher prevalence likely does not translate into higher risk of occult metastases for the most common clinical diagnoses and therefore does not support larger elective nodal treatment volumes compared with HPV-negative tumors.

Background. Progression of prostate cancer under androgen receptor (AR) pathway-targeted therapy spans a phenotypic spectrum, from tumors that sustain AR pathway output to tumors that become independent of AR signaling. Experimental models that occupy the intermediate portion of this spectrum, in which AR is still expressed but pathway output is reduced, are comparatively scarce. Methods. LNCaP cells expressing a triple-modality reporter (LNCaP-TR) were injected into nude mice by the intracardiac route, and a bone lesion that arose in the middle spine of one animal was recovered and propagated as LNCaP-TR-BoM. The subline and its parental line were compared by two-dimensional and three-dimensional culture assays, RNA-seq, qRT-PCR, and immunoblotting. AR pathway output was assessed using KLK3 expression under AR antagonist treatment. Public prostate cancer datasets provided clinical context. A CRISPR-Cas9 EPHA3-edited derivative was evaluated in aggregation and spheroid assays. Results. Relative to parental LNCaP-TR, LNCaP-TR-BoM formed more numerous and larger cell aggregates in monolayer culture and larger, less circular spheroids in both suspension and Matrigel three-dimensional culture. AR mRNA was modestly reduced and AR protein remained readily detectable, whereas KLK3 mRNA and protein were reduced disproportionately more than those of AR. Residual KLK3 expression remained suppressible by AR inhibitors enzalutamide and darolutamide. Transcriptome-based pseudotime placed LNCaP-TR-BoM between the parental line and the androgen-refractory LNCaP-RF variant. Subsequent differential gene expression (DGE) and pathway enrichment analysis revealed that LNCaP-TR-BoM exhibits transcriptomic rewiring concordant with that of clinical mCRPC, thus evidencing LNCaP-TR-BoM as a potentially clinically relevant model cell line. Finally, EPHA3 showed the largest fold change among the most significant differentially expressed genes in LNCaP-TR-BoM compared with the parental line. Therefore, EPHA3 was explored to demonstrate the utility of the LNCaP-TR-BoM cell line as a discovery system for genes that potentially drive the malignancy of bone-metastatic prostate cancer at the intermediate AR-attenuated progression stage. An EPHA3-silenced derivative of LNCaP-TR-BoM lacking detectable EPHA3 protein formed smaller aggregates and smaller, more circular spheroids, suggesting EPHA3 may play a role in driving the malignancy of AR-intermediate bone-metastatic prostate cancer. This finding provides support for the potential utility of the LNCaP-TR-BoM cell line as a discovery system for genes relevant to driving AR-intermediate bone-metastatic prostate cancer. Conclusions. LNCaP-TR-BoM is a bone lesion-derived prostate cancer cell line in which AR expression is retained but AR pathway output is attenuated, accompanied by an mCRPC-concordant transcriptomic shift and altered multicellular organization. It provides a paired discovery system for candidate genes such as EPHA3.

Purpose/Objective There is paucity of data reporting outcomes of breast cancers with initial internal mammary nodal involvement and no visceral metastases, treated with curative hypofractionated radiotherapy . We report the outcomes from a tertiary centre alongside spatial patterns of recurrences in the above group Material/Methods For this retrospective cross-sectional study, consecutive patients contoured as per the ESTRO 2013 guidelines, treated between 2016-2022 were eligible if their diagnostic imaging demonstrated involvement of the internal mammary nodes. Radiotherapy (40 Gy/15#/3 weeks) was delivered to the residual breast / thoracic wall, SCF region corresponding to the ESTRO lymph node level 4 and internal mammary chain nodes. Residual IMN/ level 4 nodes received a boost of 10Gy/5#. Spatial mapping of sites of recurrence at the local site and three nodal sites (axilla, SCF and IMN) was performed using deformable image registration. Sites of recurrence at the local site and three nodal levels were contoured separately. Volumetric intersection of the recurrent gross tumour volume (GTV_recurrence) with treated clinical target volume (CTV) was calculated. Actuarial overall (OS), disease free survival (DFS) & cumulative incidence of local (LR), regional (RR) and loco-regional recurrence(LRR) were calculated using Kaplan Meier method. Univariate comparison of outcomes with or without residual disease was performed using the log rank test. Results The median age of the 61 eligible women was 49 years. 77% received neoadjuvant chemotherapy and the rest adjuvant chemotherapy. 82% patients had a mastectomy. Axillary lymph node dissection was done in 96.7%. Boosts to residual IMN and SCF nodes were delivered to 21(34.4%) and 2 (3.3%) respectively. Median follow up was 3.6 years. Out of the 61 patients, 42 patients were disease free with an estimated 3 year disease free survival of 75% (95% CI 64, 88%). Spatial mapping of locoregional recurrence was possible in all but 1 patient with local (only) recurrence who was lost to follow-up after mammogram only. Among the patients with loco regional recurrence 1 had recurrence in local site + SCF +axilla, 3 had recurrence in the SCF+axilla, 2 in the SCF+IMN and 1 in the axilla+SCF+IMN. Only one patient had isolated axillary recurrence or isolated SCF recurrence. There were no IMN only recurrences. Among the 8 patients with nodal recurrence, a total of 27 individual GTV_recurrence were identified in the axilla(n=11), SCF(n=11) and IMN (n=5). IMN recurrences showed complete or partial overlap with CTV. SCF recurrences were a mix with predominantly in-field recurrences while axillary recurrences occurred outside the treated volume.Four (6.6%) patients had Grade 2 lymphoedema as documented late side effect. Conclusion Aggressive treatment of IMN disease with adjuvant radiation is effective with good locoregional control. Systemic recurrences are common and may benefit from intensification strategies.

Background: Patients aged [&le;]50 years with early-stage HR+/HER2- breast cancer are considered to have an aggressive disease biology and are treated with chemotherapy. However, a subset may still experience favourable outcomes without chemotherapy. Commercially available prognostic tests help guide such treatment decisions, but most are developed and validated predominantly in Western populations, with an underrepresentation of Asian patients. In this study, we explore the prognostic value of CanAssist Breast (CAB), a proteomic prognostic test, in optimal treatment management of patients aged [&le;]50 years. Methods: This study includes a previously published retrospective cohort. The performance of CAB was evaluated using Kaplan-Meier analysis, with 5-year Distant recurrence-free interval (DRFI) from diagnosis as the endpoint; the study also used multivariate analysis to evaluate the independent prognostic value of CAB. Results: In the retrospective cohort, CAB identified 70% as low-risk (LR) and 30% as high-risk (HR) with DRFI of 93.1% (P<0.0001); further classification showed 64% LR and 36% HR in the Asian and 75% LR and 25% HR in the Caucasian subgroup. In patients with N0 disease, CAB identified 85% as LR and 15% as HR. In N+ patients, CAB identified 49% as LR. All CAB LR patients have an acceptable DRFI of >90% at 5 years from diagnosis. Conclusions: Based on the results presented, CAB adds prognostic value for patients [&le;]50 years and can be used as a treatment guidance tool for these patients.

Background: Prostate cancer (PCa) is the most common urological malignancy globally and remains a major cause of cancer mortality among men. In Hong Kong, a comprehensive evaluation of disease burden, real-world treatment patterns, and outcomes across clinically relevant PCa subgroups is essential for identifying unmet needs and optimising care delivery. Objectives: This study aims to characterise the burden of PCa and patterns of care in Hong Kong by (1) estimating annual patient load (2) describing treatment patterns, and (3) evaluating real-world outcomes among patients receiving systemic therapy (chemotherapy or novel hormonal agents [NHA]) and novel agents (immunotherapy, poly ADP-ribose polymerase [PARP] inhibitors, radiation therapy). Methods: We conducted a population-based retrospective cohort study using electronic medical records from the Hong Kong Hospital Authority (HA). The cohort included patients diagnosed with PCa between January 1, 2012 and December 31, 2024. Subgroup analyses included patients with metastatic hormone-sensitive prostate cancer (mHSPC) and metastatic castration-resistant prostate cancer (mCRPC) and were further stratified by the treatment received. Descriptive analyses summarised annual patient load and treatment patterns over 2018 to 2022. We evaluated treatment outcomes from 2012 to 2024, including overall survival, progression-free survival, number of hospitalisations, and main adverse events. Results: From 2018 to 2022, prevalence increased for prostate cancer (12,014 to15,610), mHSPC (1,188 to 1,779), and mCRPC (167 to 290), while annual mCRPC incidence rose from 125 to 274. There was an increasing number of prevalent and new users of the investigated novel treatment, including first-/second-/third-line therapy for mCRPC. Compared with patients treated with NHAs, chemotherapy showed a numerically higher rate of hospitalisations, bone treatment initiation and higher hematologic toxicity. Conclusions: This population-based retrospective study provides Hong Kong-specific evidence on PCa burden, real-world management, and outcomes across key subgroups, thereby supporting policy planning, formulary evaluation, and targeted strategies to address unmet clinical needs. Funding: This work was supported by a grant from Novartis.

AbstractAutophagy and Toll-like receptor (TLR) signaling are both implicated in cancer progression, but whether they act as tumor suppressors or promoters remains unclear. To explore this relationship, we investigated the role of autophagy downstream of therapeutic TLR activation in oral squamous cell carcinoma (OSCC). Using the FDA- approved TLR agonists Bacillus Calmette-Guerin (TLR2/4) and imiquimod (TLR7), we assessed their effects in vitro on SCC-4 cells and in vivo in a chemically induced OSCC hamster model. Both agents, alone or in combination with each other or with Monophosphoryl Lipid-A induced robust autophagy, as measured by LC3B staining in vitro and flow cytometry in vivo. Autophagy induction correlated with reduced tumor volume and prolonged survival, with outcomes comparable to cisplatin, the current standard chemotherapeutic, but with less treatment-associated morbidity and mortality. Autophagy was also associated with cisplatin antitumor effects. Notably, imiquimod produced the most pronounced and sustained autophagic and antitumor effects. To our knowledge, this is the first study to directly link the therapeutic efficacy of TLR agonists in OSCC to autophagy modulation, providing both mechanistic and translational insights into their potential as immunotherapeutic agents.

Introduction: Tumors are known to develop genomic heterogeneity over time, particularly in metastatic disease, raising concerns that a single-site biopsy may be insufficient to accurately predict response to medications. Methods: We used our 3D microtumor functional profiling platform that preserves key features of the native tumor microenvironment. We tested (i) six split samples from primary tumors and (ii) seven samples from primary tumors (ovarian, colon, and breast cancer) matched with their locoregional or distant metastases. Fresh tissue specimens were processed within 24 hours after surgery and treated with NCCN guideline-recommended chemo or targeted therapies at concentrations over four orders of magnitude. Cytotoxicity was evaluated after three days to generate dose response curves. The response correlations between the split sample or between the two matched sites were quantified using the concordance correlation coefficient ({rho}c) for the drug efficacy metrics. Results: We compared a total of 275 drug responses. The six split samples resulted in a strong correlation between the two tumor areas ({rho}c=0.95, p<0.001), supporting high assay reproducibility across distinct regions of the same tumor. Locoregional metastases were highly correlated with their primaries (ovarian to omentum {rho}c=0.97, p<0.001; ovarian to liver capsule {rho}c=0.93, p<0.001; colon to peritoneum {rho}c=0.91, p<0.001). A lesser correlation was observed with lymphogenic metastases (breast to axilla {rho}c=0.87, p<0.001), and the correlations of hematogenic metastases were even lower (colon to lung {rho}c=0.69 p<0.001; ovarian to soft tissue/colon {rho}c=0.58, p<0.001). No correlation was observed among specimens that were resistant to all tested medications. Conclusion: Tumor drug responses were highly preserved in locoregional metastases, whereas somewhat greater divergence was observed in lymphogenic or hematogenic metastases. Single-site functional profiling may remain informative for identifying treatment resistance and might reduce the risk of ineffective therapy selection in metastatic disease.

Malignant peripheral nerve sheath tumors (MPNSTs) are aggressive sarcomas. An obstacle to treating MPNSTs is a lack of effective systemic therapies. Although over 70% of human MPNSTs have lost or inactivated the epigenome regulator polycomb repressive complex 2 (PRC2), its activity and contribution to canine PNST progression remain unclear. This study compared canine peripheral nerve sheath tumors (PNSTs) and human MPNSTs across biological and clinical features, including PRC2 activity. Immunohistochemical analysis was performed for a human tissue microarray of 54 neurofibromas and 139 MPNSTs, and 63 canine PNSTs for H3K27me3, a repressive histone mark deposited by intact PRC2, and H3K27ac, which increases globally upon H3K27me3 loss. To understand the genomic alterations present in canine PNSTs, we analyzed tumor mutation burden, copy number alteration, and transcriptomes of eight canine PNST/normal pairs. The results suggested that H3K27me3 loss and associated gain of H3K27ac epigenetically drive human and canine tumors. These findings warrant further studies to evaluate whether these epigenetic deregulations alter similar gene signatures across species.

Canine mammary carcinoma (CMC) exhibits heterogeneous metastatic behavior, while accurate assessment of locoregional dissemination may be limited by incomplete sampling, occult metastasis, and variable sentinel lymph node anatomy. We investigated whether primary-tumor transcriptomic programs are associated with locoregional dissemination. The training cohort comprised 113 CMC RNA sequencing samples classified by peritumoral lymphatic vessel invasion, and the independent evaluation cohort comprised 27 microarray samples classified by histopathologically confirmed lymph node (LN) metastasis. Expression profiles were mapped to human ortholog gene symbols, and single-sample gene set enrichment analysis (ssGSEA) was performed using Molecular Signatures Database (MSigDB) gene sets. Seven candidate gene sets selected through univariable screening and literature-guided curation were entered into least absolute shrinkage and selection operator (LASSO) logistic regression, yielding a four-gene-set transcriptomic risk score (RS). The RS showed an apparent receiver operating characteristic area under the curve (ROC-AUC) of 0.949 (95% confidence interval, 0.906-0.993) and precision-recall area under the curve (PR-AUC) of 0.725 in the training cohort; bootstrap internal validation yielded an optimism-corrected ROC-AUC of 0.920. In the evaluation cohort, the ROC-AUC was 0.890 (95% confidence interval, 0.766-1.000) and PR-AUC was 0.869. Discrimination was attenuated but retained in histologically restricted sensitivity analyses, and higher RS remained associated with locoregional dissemination after clinicopathologic adjustment. These findings support gene-set-level transcriptomic scoring as a potential complementary approach for characterizing dissemination-associated primary-tumor states in CMC.

Cell confluency is a major determinant of cell-cell communication, protein interactions, access to nutrients, and signalling dynamics, thereby significantly impacting biological outcomes. Lung cancer cells like A549 are widely used as screening models for scientific studies wherein their growth in vitro progress from non-confluent to confluent growth. In this study, we investigated the transcriptomic adaptations associated with the transition of A549 cells from baseline non-confluent to confluent growth. Comparative transcriptomic analysis between confluent and cells at baseline identified 815 upregulated and 671 downregulated transcripts. Pathway enrichment analysis of deregulated transcripts in confluent cells revealed enhanced cholesterol and sterol biosynthetic pathways, along with suppression of chromosomal segregation and mitotic pathways. At confluency, an increased expression of glucose transporters (SLC2, SLC60, and SL37 families) and glycolytic pathways, and a decrease in amino acid transporters (SLC1, SLC7, SLC38, and SLC36) and amino acid metabolic pathways is observed. A reduced one-carbon metabolic signature (SHMT2, DHFR, and MTHFD2) and enhanced fatty acid precursor synthesis (HMGCLL1, ALDH6A1, and AASS) were also observed at confluency. 1H NMR profiling of culture media revealed higher glucose and glutamine utilisation with lactate accumulation during culture maturation. Collectively, the data suggest transcriptome-level rewiring in A549 cells with preferential biosynthesis of lipids and sterols at confluency and underscore the importance of considering culture maturity in cancer biology, metabolism, and therapeutic studies.

Background and Objectives: Robotic-assisted natural orifice specimen extraction surgery (NOSES) is a minimally invasive approach for mid-rectal cancer, but it is technically complex and lacks a standardized operational framework. This study aimed to propose and preliminarily validate a structured modular robotic NOSES II surgical procedure for mid-rectal cancer. Methods: This was a retrospective observational study that consecutively enrolled 11 patients with mid-rectal cancer who underwent modular robotic NOSES-II surgery at our center between December 2024 and August 2025. All procedures were performed by the same surgical team strictly following the predefined six-module structured surgical protocol. Perioperative indicators, pathological outcomes, and patient-reported outcomes (PROs) at 4 to 8 months postoperatively were collected. Key techniques were analyzed via high-definition surgical videos. Results: All 11 procedures were completed successfully without conversion to open surgery. The mean operative time was 299.6 plus or minus 54.2 minutes, and the mean intraoperative blood loss was 94.1 plus or minus50.6 mL. The mean number of harvested lymph nodes was 17.4 plus or minus 8.4, with a 100% R0 resection rate. No severe complications (Clavien-Dindo grade [&ge;] III) occurred. The mean postoperative hospital stay was 8.2 plus or minus 1.5 days. Postoperative PROs indicated good defecatory function, urinary function, and overall quality of life. Conclusions: Preliminary findings demonstrate that the structured modular robotic NOSES-II procedure is safe and feasible for the treatment of mid-rectal cancer. This modular protocol provides a clear and reproducible technical framework for this complex procedure, and it is expected to achieve favorable functional preservation while ensuring oncological radicality.

PurposeThere is an urgent need for improved prognostic tools and biological understanding of chemoradiation resistance in head and neck squamous cell carcinoma (HNSCC). This study established a preclinical imaging dataset aimed at identifying prognostic imaging features from 18F-FDG micro-PET/CT scans in HNSCC mouse models. MethodsThree orthotopic murine models were utilized: two human papillomavirus (HPV)-negative (MOC1, MOC2) and one HPV-positive (MLM1). When tumor volume exceeded 50 mm3, chemoradiation was initiated using cisplatin (5 mg/kg) and image-guided radiation therapy (8 Gy) on days 0 and 7. 18F-FDG micro-PET/CT imaging was performed on day 14. Tumors were manually segmented on PET/CT, and quantitative image features including tumor volume, SUVmean, and SUVmax were extracted. Treatment response was evaluated by relative tumor size on day 11 compared to day 0. Tumor growth and survival were compared across models using multiple-effects model and log-rank. Imaging feature associations were evaluated by Mann-Whitney U tests. Associations between survival, SUVmax, and tumor volume were assessed using Cox proportional hazards modeling and Kaplan-Meier analysis with log-rank testing. ResultsA total of 121 mice were treated and imaged. Significant differences in tumor growth and survival were observed among the three models (p < 0.01 for pairwise growth comparisons; p < 0.0001 for survival). Day 11 treatment response groups demonstrated significantly different growth trajectories following chemoradiation (p < 0.0001). SUVmax was significantly associated with survival (p = 0.0009), whereas SUVmean was not significant (p = 0.13). PET tumor volume demonstrated the strongest association with survival (p < 0.0001). A multivariate Cox proportional hazards model incorporating SUVmax and tumor volume significantly stratified survival risk (p < 0.0001). ConclusionOverall, these findings demonstrate that 18F-FDG PET/CT-derived metrics, particularly SUVmax and tumor volume, are robust predictors of chemoradiation response in orthotopic murine models of HNSCC.

Purpose: There is an unmet need in the treatment of triple-negative breast cancer (TNBC), as recurrence rates remain high; thus, novel treatment strategies are warranted to improve outcomes for patients. Smac mimetics (SMs) are small molecules that inhibit the inhibitors of apoptosis. In this study, we investigated the properties of the SM Xevinapant (AT406) as a radiosensitizer in TNBC. Methods: TNBC cell lines were treated with Xevinapant (AT406) in the presence or absence of ionizing RT and cell viability and apoptosis measured in vitro. Clonogenic assays were performed to confirm radiosensitization. We also assessed xevinapant radiosensitization in the presence of the Tumor Necrosis Factor (TNF) cytokine, which has been shown to enhance SM-induced apoptosis. Radiosensitization mechanisms were further validated by western blotting, flow cytometry and cell cycle analysis. Results: We show that Xevinapant increased cell death in a panel of irradiated TNBC cell lines. In clonogenic assays, Xevinapant sensitized MDA-MB-231 cells to RT-induced cell death, and this was significantly enhanced in the presence of TNF, with a dose-enhancing factor (DEF) of 1.57. We also show that Xevinapant radiosensitization is mediated by TNF-dependent apoptosis. Conclusions: Xevinapant is a radiosensitizer that enhances RT-induced cell death in TNBC cells, and TNF is central to this process through activation of apoptosis. These results are promising for combining RT with Xevinapant to improve local control in TNBC.

Background: Objective assessment of tumor burden in patients with solid tumors remains inexact. Furthermore, while mutation-based liquid biopsies are specific, they are limited by tumor heterogeneity and the requirement for detectable clonal mutations. Cell-free DNA (cfDNA) concentration, a cost-effective substrate for mutation-focused liquid biopsy, has shown promise as a "molecular tumor burden" biomarker. Methods: We analyzed cfDNA concentration from 1000 unique cancer patients using standardized protocols. Demographics, AJCC stage, clinical anatomic tumor burden and oncologic outcome variables were collected. Associations were assessed with non-parametric tests. Multivariable linear regression and Cox proportional hazards models were used to identify independent predictors and survival associations. Results: CfDNA concentration varied broadly (median 6.25 ng/mL; range 0.5-1132.9). Anatomic tumor burden, including tumor number (rho=0.25, p<0.0001) and largest tumor diameter (rho=0.25, p<0.0001), correlated significantly with cfDNA, especially in stage IV disease. Primary tumor site influenced cfDNA levels, with liver/bile duct cancers having higher cfDNA than other sites (median 13.5 vs 6.1 ng/mL, p<0.0001). Multivariable analysis confirmed overall tumor burden and hepatic tumor location (primary or metastatic) as principal drivers of cfDNA. Critically, cfDNA concentration was an independent predictor of shorter PFS (p=0.001) and DSS (p<0.0001), demonstrating increased value in advanced disease settings, irrespective of treatment intent. Conclusions: CfDNA concentration is a robust, biologically integrated biomarker that provides an objective measure of total systemic tumor burden and prognosis in a large, pan-cancer cohort. By capturing disease activity irrespective of mutational status, it offers a valuable adjunct to targeted profiling, particularly in heterogeneous or advanced malignancies. Although these findings require prospective clinical validation, cfDNA concentration may eventually augment patient selection for aggressive versus palliative interventions, especially in advanced disease.

Background: Prostate cancer enzalutamide treatment is approved at a standard dose of 160 mg daily. Concerns for real-world patients -- older and more fragile than those enrolled in clinical trials -- have prompted consideration of initiating treatment with lower doses, but the long-term efficacy of this approach remains unknown. We evaluate the long-term survival and longevity in patients treated with standard versus upfront low-dose enzalutamide. Methods: Retrospective analysis of 151 patients treated with enzalutamide (102 receiving 160 mg; 49 receiving [&le;]80 mg) between 2014--2021 at the Centre Hospitalier Universitaire de Martinique, with complete follow-up through end of life (98.7% completeness of follow-up). Primary outcomes were overall survival (OS), progression-free survival (PFS), and longevity (attained age). Results: Doses [&le;]80 mg were associated with longer median OS (36.3 vs. 20.7 months), improved restricted mean OS (difference of 0.7 years, p=0.05), and enhanced longevity (median 82.5 vs. 78.3 years, p=0.004). PSA response rate at 12 weeks was higher with lower-dose (71.4% vs. 48.8%, p=0.016). In multivariable models adjusted for prognostic factors, [&le;]40 mg compared with 160 mg was non-inferior regarding OS (HR=0.61, 95% CI 0.36--1.06), superior regarding PFS (HR=0.59, 95% CI 0.35--0.99), and superior regarding longevity (HR=0.48, 95% CI 0.28--0.84). Bone metastasis, poor performance status, PSA response, time to PSA nadir, and disease duration were independent predictors of outcomes. A post-hoc analysis revealed a strong association between dose and physician-prescribing profiles, ranging from "endorse-lowest-dose" to "never-deviate-from-full-dose". Conclusions: Lower doses of enzalutamide were non-inferior to full-dose. Dose-adapted strategies warrant further investigation.

Non-muscle invasive bladder cancer (NMIBC) exhibits high recurrence, necessitating frequent invasive surveillance. Urine DNA methylation profiling represents a promising non-invasive biomarker approach for early detection and monitoring. This pilot study aimed to investigate urine DNA methylation patterns in NMIBC among Thai patients. Five patients with high-grade NMIBC and three healthy volunteers were enrolled, and midstream urine and blood samples were collected prior to transurethral resection of bladder tumor (TUR-BT). Urine cell-free DNA (cfDNA) was extracted, mechanically sheared from an atypical peak of approximately 400 bp to an optimal target length of 150-200 bp, and analyzed using cell-free methylated DNA immunoprecipitation followed by high-throughput sequencing (cfMeDIP-seq). Differentially methylated regions (DMRs) were identified between NMIBC and controls, and principal component analysis (PCA) was performed. A total of 82 DMRs were identified (adjusted P < 0.001), including 76 hypermethylated and 6 hypomethylated regions, with IRX4, TBX5, and MIR124-3 representing the top hypermethylated genes. To account for cohort heterogeneity, a refined subset analysis excluding two outlier samples (UC2 and UC3) was performed, revealing 476 significant DMRs (449 hypermethylated). PCA demonstrated clear, robust segregation between NMIBC and controls, highlighting MIR124-3, IRX4, and LHX1-DT as the most significantly hypermethylated targets. Our findings demonstrate that urine cfDNA methylation profiling can robustly distinguish NMIBC from healthy controls, highlighting MIR124-3, IRX4, and LHX1-DT as highly promising, demographic-specific biomarkers. This non-invasive approach successfully bypasses the degradative limits of traditional bisulfite conversion and establishes a technically validated foundation for clinical surveillance and diagnostics

Purpose The Karolinska Liver Metastases (KaroLiver) cohort was established to investigate associations between clinical characteristics and histopathological features in patients treated with curative intent for colorectal cancer liver metastases (CRLM). The cohort combines whole-slide digital histopathology images with detailed oncological, surgical, radiological and survival data, enabling comprehensive analyses of treatment trajectories, clinical outcomes and metastatic tumour biology. Participants KaroLiver is a retrospective observational cohort comprising all consecutive patients who received curative-intent, liver-directed treatment for CRLM at Karolinska University Hospital in Stockholm, Sweden. The hospital is the primary regional referral centre for HPB surgery, serving the population of approximately 2.5 million people in the Stockholm-Gotland healthcare region. Patient enrolment is continuously updated in accordance with amended ethical approvals and evolving scientific questions. The cohort currently comprises 811 patients who underwent 1204 liver interventions between February 2012 and January 2022. Detailed clinical, oncological, surgical, pathological, molecular, recurrence and survival data are collected. Findings to date Median overall survival (OS) in the current cohort is 51.0 months (95% CI 46.2-57.1 months), and median recurrence-free survival (RFS) is 10.4 months (95% CI 9.2-11.9 months). The five-year OS rate is 44.9% (95% CI 41.2-48.8%). Studies using the cohort have so far identified a liver injury-derived stromal capsule in a subset of metastases, associated with improved survival. The cohort has also enabled the identification of histopathological markers of tumour biology, sex-based differences in treatment and survival, and associations between post-hepatectomy liver failure and oncological outcomes. Future plans Current research priorities include advanced histology-based prognostic scoring, sex differences in recurrence and retreatment, tumour biology and outcomes in early-onset versus average-onset CRLM, as well as CT- and MRI-based radiomics, all integrated within KaroLiver's histopathological framework. Data sharing is supported, given that regulatory requirements are met. Retrospective accrual and outcome updates will continue for current and future studies, subject to the required approvals.

Background & Aims: Cholangiocarcinoma (CCA) is an aggressive malignancy with poor five-year survival. Improving outcomes requires preclinical models that faithfully recapitulate the native tumour microenvironment. Precision-Cut Tissue Slices (PCTS) retain in-vivo architecture and represent a promising candidate. We aimed to establish a patient-derived PCTS model of CCA, characterise its response to ex-vivo culture, and assess its use as a platform for therapeutic testing. Methods: PCTS were generated from 25 patients undergoing curative-intent resection for presumed CCA (2022-2024) and cultured for up to 15 days. Viability and histological architecture were assessed serially. Quantitative proteomics (SWATH-DIA) was performed at Days 0, 3, 7 and 15. A subset were treated with staurosporine or clinically relevant chemotherapy (5'-deoxy-5-fluorouridine, gemcitabine {+/-} cisplatin). Results: PCTS viability was maintained to Day 15, with no significant reduction from baseline. PCTS retain tumour architecture, cytokeratin-19-positivity and resident CD3+/CD68+ immune cells during culture. Proteomic profiling quantified 4,578 proteins and identified a staged response, with differentially abundant proteins increasing from 8 (Day 3) to 127 (Day 7) and 180 (Day 15), stabilising thereafter. This comprised of an early loss of inflammatory and stromal proteins, sustained cellular stress responses and metabolic reprogramming. CCA subtype-specific proteomic signatures were preserved throughout culture. Staurosporine produced dose-dependent cytotoxicity whilst chemotherapy responses were variable. Conclusions: Patient-derived CCA PCTS maintain viability, tumour architecture and subtype identity for 15 days while undergoing a defined proteomic response to culture, and retain pharmacological responsiveness,

Background: Small cell lung cancer (SCLC) is an aggressive, high-grade neuroendocrine carcinoma (hgNEC) with poor prognosis. Tarlatamab, a DLL3-targeting T cell engager was approved in 2024 for relapsed SCLC, with almost doubled overall survival benefit compared to chemotherapy. However, over half of patients do not respond to tarlatamab and others develop resistance within months. With multiple DLL3-targeting therapies in clinical trials, there are currently no validated predictive biomarkers to identify patients most likely to benefit from tarlatamab. In this study, we evaluate baseline DLL3 IHC level in patients as a predictive biomarker to tarlatamab clinical outcome. Methods: We assembled a cohort of 138 patients within IRB-approved MD Anderson GEMINI database (PA13-0589) with DLL3 expression by CLIA-validated immunohistochemistry (IHC), and those treated with tarlatamab from 7/1/2024 to 3/30/2026. Only those with DLL3 IHC results and treated with tarlatamab were included in this predictive biomarker study. All DLL3 levels were reported as percentage cells with cytoplasmic or surface DLL3 expression, including 24 with additional intensity score and H-score calculation. demographic, clinical, and outcome data were collected. Systemic and intracranial response data were assessed by RECIST and mRANO BM criteria respectively. Longitudinal liquid biopsies were taken from a subset of patients for circulating tumor DNA (ctDNA) and (Cytometry by Time-of-Flight) CyTOF analyses. Results: In the predictive biomarker cohort of 54 patients with SCLC treated with tarlatamab, Median DLL3% level in tarlatamab treated cohort was 75%. Median follow-up time was 9.9 months, median time on tarlatamab treatment (ToT) was 5.8 months. At the data cut-off, 61% (33 out of 54) patients were alive. Kaplan-Meier curve stratified by DLL3 % of <=75% vs. >75 showed significant longer ToT and a trend toward longer median OS (mOS). In DLL3>75% compared to DLL3 <=75%: mToT was 10.1 months vs. 2.9 months (p=0.036), mOS: not reached vs. 9.5 months (p= 0.066). Among 41 patients who completed one cycle of tarlatamab treatment and were eligible for systemic (extra-cranial) response per RECIST criteria; we observed partial response (PR) in 51% (21) patients, stable disease (SD) in 22% (9) patients; and progressive disease (PD) in 27% (11) patients. The median DLL3 % in those with PR vs. PD were 80% vs. 40% (p=0.006); median DLL3% comparing PR and SD were 80% vs. 60% (p=0.051). Among 35 patients evaluable for intracranial response, best objective response prior to any brain radiation were 5 (14%) CR,11(31%) PR, 8 (23%) SD and 11 (31%) PD. 9 received brain radiation (8 SRS, 1 WBRT) during tarlatamab to achieve better intracranial control while systemic control is maintained. DLL3% does not appear to be correlated with intracranial response. Longitudinal ctDNA and CyTOF analyses showed persistent ctDNA positivity, as well as increasing NEUROD1 subtype cells in circulating tumor cells in post-tarlatamab progression samples. We additionally profiled DLL3 expression by IHC in 138 patients with SCLC and found heterogenous DLL3 expression, with Median DLL3% of 70% across different biopsy sites. Conclusions: Our data suggests DLL3 expression in patients with SCLC is heterogeneous and not ubiquitous, with median DLL3 IHC of 70%. DLL3 IHC predicts objective response and long-term outcome with tarlatamab. In addition, dynamic monitoring of ctDNA level and CTCs by CyTOF are suggestive of development of tarlatamab resistance, however further studies with larger cohort of paired longitudinal samples are needed to validate the novel blood-based biomarkers. To our knowledge, this study is the first to validate DLL3 IHC as a predictive biomarker for tarlatamab in SCLC, which leads the way for optimizing treatment selection and combinatorial therapies for the subset of patients less likely to respond to tarlatamab.