Cancers

Introduction Stark disparities in endometrial cancer (EC) risk and mortality exist between non-Hispanic Black and White women, with Black women experiencing higher incidence and worse survival. This disparity has been attributed to biological and socioeconomic factors, though how these factors interact to influence EC disparities remains unclear. This study modeled EC outcomes using race, area-level socioeconomic deprivation, clinical phenotypes, genetic ancestry, and molecular alterations. Methods We identified 281 cases of EC diagnosed from 2013-2023 in women who underwent clinical genomic sequencing as part of routine care across multiple Northwell Health sites. We estimated genetic ancestry, oncogenic alterations in 324 genes, microsatellite instability, and molecular classification. Geocoded patient addresses were used to derive the state-level Area Deprivation Index to estimate socioeconomic deprivation. Results African ancestry patients were enriched for high-grade disease (89% vs 64%), serous histology (57% vs 26%), and the TP53-mutant molecular classification (71% vs 51%) compared to European ancestry patients (p-value<0.05). Socioeconomic deprivation quintiles were associated with race, with more deprived quintiles enriched for Black patients (p-value<0.001). Both race and genetic ancestry, but not area-level deprivation, were independently associated with differences in progression-free survival. TP53 mutations were enriched in African ancestry patients, while KRAS, PTEN, and ARID1A mutations were enriched in European ancestry patients (q<0.10). Cox proportional hazards modeling, adjusting for these factors, showed that African ancestry patients had worse progression-free survival (HR 1.91, p-value<0.05). Conclusion Our findings indicate that EC disparities persist after adjusting for socioeconomic, clinical, and molecular factors, highlighting the need to further investigate additional drivers of disparity.

PURPOSE Colorectal cancer (CRC) incidence and mortality rates differ by population, and evidence suggests that genetic differences may affect cancer biology. However, studies investigating CRC variants in genetically heterogeneous populations are limited. Using somatic tumor mutation profiling of CRCs diagnosed in African Americans (AAs), Ghanaians, Ethiopians, and NHWs, we explore correlations between population group and population-specific tumor variants. PATIENTS AND METHODS Somatic DNA from CRC tumors resected from 150 individuals, including 43 AAs (27%), 53 NHWs (35%), 21 Ghanaians (14.2%), and 33 Ethiopians (22.3%), was sequenced on the Illumina NovaSeq platform, targeting 290 genes. We compared mutations in AAs, Ghanaians, and Ethiopians to those in NHWs to identify variants enriched in historically underrepresented groups. RESULTS US cohort tumors were diagnosed at significantly younger ages with more early-onset cases (<50 years old) than African cohorts (p <0.05). Significant differences were observed in primary tumor location, MMR phenotypes, KRAS mutations, and distribution of tumor mutational burden by population. BRAF V600E mutations were rare across all groups, while non-V600E BRAF mutation rates were higher in AA and NHW (43-44%) than Ethiopian and Ghanaian (14-33%) samples. Population-specific differences were identified in mutation rates of APC, CTNNB1, RNF43, PIK3CA, and TP53, as well as in pathogenic variant occurrence.

Metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) is a long non-coding RNA (lncRNA) implicated in cancer progression. In thyroid cancer, MALAT1 has been proposed as a potential biomarker, but its role in disease progression remains incompletely understood. Here, we analyzed MALAT1 RNA levels in paired tumoral and adjacent non-tumoral thyroid samples from a Chilean patient cohort. We found a positive correlation of MALAT1 levels with lymphatic infiltration that was not replicated when modeling MALAT1 expression in a larger cohort obtained from the TCGA-THCA database. An exploratory RNA-seq comparison of one matched tumor-adjacent tissue pair confirmed higher tumor abundance of MALAT1 and the epithelial-to-mesenchymal transition-marker VIM, together with lower abundance of cell-adhesion gene PCDH10. To investigate the impact of MALAT1 on thyroid cancer and cellular metabolism, we targeted MALAT1 in the papillary thyroid cancer cell line TPC1. MALAT1 knock-down reduced proliferation and migration while enhancing mitochondrial respiration with no changes in glycolysis. Notably, although MALAT1 was not localized within mitochondria, its silencing modulated the expression of transcripts associated with mitochondrial dynamics and mitophagy. Consistent with these results, transcriptomic correlation analysis in the TCGA-THCA cohort showed that MALAT1 expression was largely uncoupled from oxidative phosphorylation and glycolysis gene programs, while negatively correlating with core regulators of mitophagy and mitochondrial dynamics, pointing to a link with mitochondrial quality control rather than direct bioenergetic reprogramming. Our findings highlight MALAT1 as a contributor to thyroid cancer aggressiveness and reveal a link between MALAT1 and mitochondrial quality control independent of direct mitochondrial localization. Besides, our results support a tissue-specific mechanism and population-specific role of MALAT1 in cancer biology.

BACKGROUND: Anemia is a negative factor in cancer, influencing the prognosis, quality of life and financial situation of cancer patients. Recent studies have shown that anemia in cancer is provoked by augmented erythrocyte removal. OBJECTIVE: In this study we sought to investigate the molecular bases for erythrocyte removal in cancer patients with anemia. In particular, we explored the levels of erythrocyte CD47, lactadherin, calreticulin and MCP1. METHODS: Thirty five anemic cancer patients (25 women, aged 66.4 +/-11.35 years old) and twelve healthy non-anemic controls (8 men, aged 61.1+/-9.98 years old) participated in our study. Red blood cells were isolated throug multiple centrifugations, and were lysed with the use of Triton-X 100. The levels of CD47, lactadherin, calreticulin and monocyte chemoattrractant protein 1 were determined by ELISA. RESULTS: Erythrocytes of anemic cancer patients display reduced CD47 (p<0.001), MCP1 (p=0.05), CD47 to lactadherin ratio (p<0.05), and increased lactadherin levels (p<0.01) in comparison to the healthy controls. DISCUSSION: Reduced CD47 along with increased lactadherin possibly drive erythrocyte removal in anemic cancer patients. Conclusions: The role of CD47 and increased lactadherin should be examined in the future as potential therapeutic targets and biomarkers for anemia diagnosis and iron dymsetabolism.

Background: Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype characterized by the absence of estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2 expression. It is associated with limited targeted treatment options, early relapse, and a high propensity for visceral metastasis. Data describing metastatic patterns and treatment characteristics of TNBC in Nigeria remain limited. Methods: This retrospective descriptive cohort study included 869 patients with TNBC managed at the Medserve-LUTH Cancer Center, Lagos University Teaching Hospital, Nigeria between June 2019 and June 2024. Demographic, clinicopathologic, metastatic, and treatment-related data were extracted from electronic medical records. Descriptive statistics were used to summarize patient characteristics, metastatic patterns, and treatment profiles. Associations between metastatic disease and selected clinicopathologic and treatment variables were explored using Pearsons chi-square test. Complete-case analysis was applied throughout. Results: The mean age at presentation was 52.09 {+/-} 12.26 years. Most patients were married (79.1%), postmenopausal (64.3%), and of Yoruba ethnicity (56.8%). Advanced disease predominated, with Stage III and Stage IV disease accounting for 42.9% and 35.6% of cases, respectively. Invasive ductal carcinoma was the most common histologic subtype (77.0%), while Grade II tumours constituted 51.3% of graded cases. Surgery was performed in 73.1% of patients, predominantly mastectomy (70.9% of surgical procedures). Chemotherapy was administered to 83.2% of patients, most commonly anthracycline-based regimens (41.8%), while radiotherapy was delivered to 63.5% of patients, with hypofractionated schedules of 42-43 Gy in 15-16 fractions accounting for 47.2% of radiotherapy courses. Metastatic disease was documented in 32.9% of evaluable patients. Lung metastasis was the most frequent site (62.5%), followed by bone (46.3%), regional lymph node invasion (38.5%), liver (23.0%), and brain (22.6%). Tumour grade and histologic subtype were not significantly associated with metastatic disease, whereas radiotherapy exposure demonstrated a significant association with metastatic status ({chi}{superscript 2} = 10.35, p = 0.001). Conclusion: TNBC in this Nigerian cohort was characterized by advanced-stage presentation, invasive ductal predominance, extensive use of multimodality treatment, and substantial visceral metastatic burden. Lung metastasis was the most common metastatic site. These findings provide contemporary real-world data on TNBC in Nigeria and highlight the continuing need for earlier diagnosis, timely referral, and sustained investment in comprehensive cancer care services.

T2, an N4-aryl-substituted thiosemicarbazone, has previously been shown to exert cytotoxic and anti-invasive effects in triple-negative breast cancer (TNBC) and to increase expression of the metastasis suppressor N-myc downstream-regulated gene 1 (NDRG1). Given the role of NDRG1 in regulating epithelial-mesenchymal transition (EMT) and WNT/{beta}-catenin signaling, we investigated the contribution of this pathway to the anti-invasive activity of T2. The effects of T2 on WNT/{beta}-catenin signaling and associated microRNAs (miR-182-5p and miR-200c) were evaluated in 4T1 cells. In vivo activity was assessed using a fully immunocompetent intraductal 4T1 mouse model that recapitulates the progression from ductal carcinoma in situ (DCIS) to invasive ductal carcinoma (IDC). Tumor progression, invasion, NDRG1 expression, and WNT/{beta}-catenin pathway components were analyzed. T2 reduced WNT/{beta}-catenin signaling and modulated the expression of miR-182-5p and miR-200c in vitro. In the MIND model, T2 decreased the frequency of invasive lesions and reduced {beta}-catenin, ZEB1, and c-Myc expression while increasing NDRG1 levels. {beta}-catenin localization differed between lesion types, showing predominantly membrane-associated staining in DCIS lesions and a diffuse cytoplasmic distribution in invasive foci. These findings identify WNT/{beta}-catenin signaling and NDRG1-associated pathways as potential mediators of the anti-invasive effects of T2 in TNBC. The reduction in invasive progression observed in the MIND model supports further investigation of this compound in preclinical models of TNBC.

P2Y12 receptor (P2RY12), mainly expressed on platelets, is known for its central role in hemostasis. P2RY12 activation is also involved in cancer development through platelet adhesion to cancer cells supporting immune-evasion, promoting tumor angiogenesis and metastasis, among others. P2RY12 is known as an actionable target, and P2RY12 antagonists are in clinical use for cardiovascular diseases. However, very little data are available regarding the protein expression of P2RY12 in lung carcinomas. We performed immunohistochemical staining for P2RY12 in a cohort of non-small cell lung cancer (NSCLC) samples comprising 320 adenocarcinomas (LUAD) and 158 squamous cell carcinomas (LUSC). Results were evaluated using a dual approach combining microscopic assessment and digital image analysis (QuPath). Results were correlated with clinical-pathological data. We found significantly higher P2RY12 protein expression in LUSC compared to LUAD (p<0.001) via eyeballing (absent/low expression in 21.7% (34/158) and moderate/high expression in 78.3% (124/158) of LUSC cases versus absent/low expression in 98.4% (315/320) and moderate/high expression in 1.6% (5/320) of LUAD cases). Digital analysis yielded similar results. High P2RY12 expression was associated with a significantly better 5-year overall survival rate for the entire cohort (p=0.0048) as well as for the LUAD (p=0.015) and LUSC (p=0.05) subgroups. Furthermore, P2RY12 showed excellent discriminatory performance for classifying carcinomas as LUAD or LUSC, with an AUC of 0.916 in ROC-analysis. High P2RY12 expression is linked to a better prognosis and might serve as a promising novel prognostic biomarker for NSCLC. Its assessment could be implemented in future routine diagnostic workup. At the same time, the data suggest that P2RY12 could also serve as a diagnostic marker for LUSC.

Background: Cutaneous squamous cell carcinoma (cSCC) accounts for a significant proportion of skin malignancies in India, yet data on patterns of failure, particularly for extremity and truncal primaries remain scarce. We audited a decade of surgically treated cSCC at a tertiary cancer center to characterize failure patterns and associated risk factors. Methods: This retrospective study included 161 patients with histopathologically confirmed cSCC treated surgically between January 2013 and December 2023, comprising 127 upfront/residual and 34 recurrent presentations. Primary sites were extremities (64%), head and neck (26%) and torso (10%). 21 patients had Marjolin's ulcer. Outcomes included local, regional and distant failure, recurrence-free survival and overall survival. Brigham and Women's Hospital (BWH) staging was applied to assess prognostic utility. Statistical analysis was done using Kaplan-Meier and competing-risk methods. Results: Median follow-up was 2.4 years. Regional recurrence was the predominant failure pattern seen in 26 patients, local recurrence was seen in 14 patients and distant metastasis in 13. The 3-year cumulative incidences of local, regional and distant failure were 11%, 19% and 8.4% respectively. Rates of regional recurrence were substantially higher than Western series. Extremity primaries accounted for 19/26 regional recurrences. BWH T2b disease showed the highest regional failure rate (27.6%), exceeding T3 (17.8%) and T2a (6%) with perineural invasion significantly associated with regional failure in T2b/T3 tumors (p<0.001). Median time to regional metastasis was 8.4 months. At 3 years, overall survival was 77% and progression-free survival was 64%. Conclusion: Regional recurrence is the dominant mode of failure in this cohort, at rates higher than most published series, with extremity primaries and BWH T2b staging identifying particularly high-risk subgroups. These findings highlight the need for a comprehensive staging system encompassing non head and neck cSCC and support prospective evaluation of elective nodal staging and adjuvant radiotherapy in high-risk patients, alongside intensified surveillance.

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.

Objective: The purpose of this study is to determine how family history of cancer, genetic mutations, presenting symptoms, and comorbidity burden collectively influence cancer outcomes in patients with epithelial ovarian cancer. Methods: A retrospective analysis was conducted on all patients with epithelial ovarian cancer treated at the University of Missouri and Ellis Fischel Cancer Center between 2008 and 2024. Patient charts were reviewed for histological subtypes, stage of cancer, status of metastasis, CA-125 values, presenting symptoms, comorbidities, family history of cancer, genetic mutations, and survival outcome. Cox regression and association analyses were performed. Results: In this cohort of patients, comorbidities and genetic mutations did not influence ovarian cancer survival. While histological subtypes, CA-125 levels, and cancer stage remained strongly associated with survival. Significant associations were observed between certain presenting symptoms and cancer histological subtype, a family history of breast cancer, stage of cancer at diagnosis, the status of metastasis, and CA-125 levels. Conclusion: Comorbidities and genetic mutations were not significantly associated with ovarian cancer survival. Presenting symptoms were associated with several clinical and pathological variables linked to ovarian cancer diagnosis.

Abstract Background: The lack of effective biomarkers and therapeutic targets to overcome radioresistance in cervical cancer remains a major clinical challenge. Tumor necrosis factor receptor-associated factor 6 (TRAF6), an E3 ubiquitin ligase pivotal in immune and inflammatory signaling, has been implicated in various malignancies. However, its role in radioresistance in cervical cancer remains unclear. Methods: TRAF6 expression was evaluated in cervical cancer tissues from 162 patients who underwent postoperative radiotherapy at our institution and in 304 cases from the TCGA-CESC cohort. The prognostic significance of TRAF6 was assessed using Kaplan-Meier and Cox regression analyses. A nomogram integrating TRAF6 expression with clinicopathological factors was constructed to predict overall survival (OS) and progression-free survival (PFS). The functional role of TRAF6 in malignant phenotypes and radiosensitivity was investigated using shRNA-mediated knockdown in HeLa and C33A cervical cancer cells. Immune cell infiltration patterns associated with TRAF6 expression were analyzed using ssGSEA and xCELL algorithms based on TCGA data. Results: TRAF6 expression was significantly elevated in cervical cancer tissues compared with adjacent normal tissues (70.99% vs. control, P < 0.001) and was higher in radioresistant than in radiosensitive patients (P < 0.001). High TRAF6 expression was associated with shorter OS (HR = 18.73, P = 0.004) and PFS (HR = 8.44, P < 0.001) and was identified as an independent risk factor for radiotherapy resistance (OR = 8.44, P < 0.001). The TRAF6-integrated nomogram demonstrated good predictive accuracy for OS (C-index = 0.7351) and PFS (C-index = 0.7444). TRAF6 knockdown in cervical cancer cells significantly suppressed proliferation, migration, and invasion, while substantially enhancing radiosensitivity of tumor cells. Functional enrichment analysis revealed that TRAF6-related genes were enriched in autophagy, mitophagy, and HPV infection pathways. Immune cell infiltration analysis showed that TRAF6 expression correlated with distinct immune cell profiles, characterized by enrichment of activated dendritic cells, M1 macrophages, and regulatory T cells, alongside depletion of cytotoxic effectors such as CD8+ T cells and {gamma}{delta} T cells. Conclusions: TRAF6 could be a prognostic biomarker associated with poor outcomes and indicator of radiotherapy resistance in cervical cancer, TRAF6 represents a potential therapeutic target for overcoming radioresistance in cervical cancer.

Melanoma is an aggressive malignancy that rapidly adapts to therapy. While chemotherapy resistance has traditionally been attributed to tumour-intrinsic mechanisms, growing evidence implicates the tumour microenvironment in shaping drug tolerance. However, few in vitro models capture the stromal complexity needed to study this interaction. We developed two multicellular melanoma spheroid models of increasing stromal complexity: a baseline model of melanoma, endothelial, and macrophage cells (BEM), and a fibroblast-containing counterpart (BEMF), and compared their transcriptional response to doxorubicin. Fibroblast inclusion increased the doxorubicin concentration required to achieve comparable growth inhibition. While untreated BEMF spheroids exhibited only modest baseline transcriptional differences, they showed a profoundly reshaped transcriptional response after doxorubicin exposure, displaying broader and higher-magnitude changes. These responses were characterized by suppression of proliferative and cell-cycle programmes, together with activation of inflammatory, immune-associated, metabolic, and stress-adaptive pathways. Higher-resolution pathway analyses further revealed coordinated attenuation of mitotic progression, checkpoint regulation, homologous recombination repair, and Rho GTPase signalling, consistent with a shift toward stress-adaptive and phenotypically plastic states, rather than classical resistance mechanisms. Transcriptome-derived transcription factor activity inference supported this regulatory rewiring. Integration with curated resistance-associated genes and external transcriptomic datasets demonstrated strong conservation of core transcriptional features across heterogeneous experimental systems, including consistent suppression of proliferation-associated genes and induction of inflammatory signalling programmes. Together, these findings indicate that fibroblasts redirect chemotherapy responses toward a stress-adaptive, persister-like phenotype and establish fibroblast-containing 3D melanoma spheroids as a physiologically relevant platform for studying tumour microenvironment-mediated chemotherapy tolerance and stromal-tumour interactions.

Background Zinc finger protein 36-like 1 (ZFP36L1) is an AU-rich element-binding RNA-binding protein that regulates post-transcriptional gene expression and has been implicated in tumor progression, cell-cycle regulation, and DNA damage responses. However, its functional role in triple-negative breast cancer (TNBC) remains poorly understood. This study investigated the effects of CRISPR/Cas9-mediated ZFP36L1 knockout on cell proliferation, doxorubicin (DOX) sensitivity, cell-cycle progression, and DNA damage responses in MDA-MB-231 TNBC cells. Methods Wild-type (WT) and CRISPR/Cas9-generated ZFP36L1 knockout (KO) MDA-MB-231 cells were cultured under standard conditions. Cellular proliferation was evaluated by cell counting over three weeks. Cell viability following DOX treatment was determined using the MTT assay, and half-maximal inhibitory concentration (IC50) values were calculated. Cell-cycle distribution was assessed by propidium iodide flow cytometry after 24 h of DOX exposure, while DNA damage was quantified by {gamma}-H2AX flow cytometric analysis. Statistical significance was determined using Student's t-test with P < 0.05 considered significant. Results ZFP36L1 knockout reduced the proliferative capacity of MDA-MB-231 cells compared with WT cells. Both cell lines exhibited dose-dependent decreases in viability following DOX treatment. KO cells demonstrated a higher mean IC50 than WT cells (9.64 vs. 8.40 M), indicating a trend toward reduced DOX sensitivity; however, this difference was not statistically significant (P = 0.569). Flow cytometric analysis revealed enhanced accumulation of KO cells in the S and G2/M phases following DOX treatment, suggesting altered cell-cycle checkpoint regulation. Furthermore, KO cells exhibited elevated basal {gamma}-H2AX expression and greater DOX-induced {gamma}-H2AX accumulation than WT cells, indicating increased DNA damage and impaired maintenance of genomic stability. Conclusions CRISPR/Cas9-mediated loss of ZFP36L1 suppresses proliferation, alters cell-cycle checkpoint dynamics, and enhances DNA damage accumulation in MDA-MB-231 TNBC cells. These findings indicate that ZFP36L1 plays a context-dependent role in regulating genomic stability and cellular responses to genotoxic stress, highlighting its potential as a biomarker and therapeutic target in triple-negative breast cancer.

Treatment resistance and metastases occur in 10-20% of patients with invasive lobular carcinoma (ILC), the most common special histological subtype of breast cancer. ILC metastasizes to the ovary more frequently than no special type (NST) tumors, also known as invasive ductal carcinoma (IDC). To characterize the genomic landscape of breast cancer ovarian metastases, we analyzed 15,613 local breast cancers, 22,010 non-ovarian metastases, and 246 ovarian metastases sequenced using FoundationOne(R)CDx or FoundationOne(R) assays. Ovarian metastases had enriched CDH1, PIK3CA, and TBX3 mutations and depleted TP53 and MYC alterations relative to local breast cancers, with additional depletion of ESR1 mutations compared to non-ovarian metastases. CDH1 mutations were less frequent in ovarian metastases (47%) than local ILC (81.3%), with reduced 16q loss (64% vs 84%), indicating that ovarian metastases also arise from non-ILC tumors. We extended these findings to a UPMC cohort of 27 ovarian metastases (13 ILC, 8 IDC, 6 mixed ductal-lobular carcinoma) with patient-matched primary tumors in most cases. In both cohorts, patients with ovarian metastases were significantly younger than those with other metastatic sites. In the UPMC cohort, the most frequent mutations were in PIK3CA, CDH1, KMT2C, FOXA1, and RUNX1. Transcriptomic analysis identified upregulated G protein-coupled receptor (GPCR) pathways, including metabotropic glutamate receptor signaling. Functional studies showed that calcium-sensing receptor (CaSR), a GPCR overexpressed in ovarian metastases, drives MEK/ERK-dependent migration and F-actin reorganization in ILC cell lines, enhanced by estrogen and blocked by calcilytic, MEK, or anti- estrogen treatment. Our findings inform future therapeutic targeting of ovarian metastasis.

Background: Despite advances in circulating tumor DNA analysis, reliable detection of oncological disease from ultra-low coverage whole genome sequencing (ulcWGS) remains challenging, particularly at low tumor fractions. This study leverages cell-free DNA (cfDNA) characteristics to develop and evaluate a robust, integrative binary predictive model for ovarian cancer (OC) status screening. OC represents a growing global burden and is often diagnosed at advanced stages due to the lack of specific early symptoms and effective screening strategies, highlighting the need for sensitive and broadly applicable early detection approaches.Methods and Findings: We analyzed plasma cfDNA from OC patients (N = 85) and cancer-free controls (N = 41) using ulcWGS (~1x). Participation in the study was voluntary, and all participants provided written informed consent before any study-related procedures under study approval No. 16119-8/2022/EUIG. Within an integrated workflow combining standardized laboratory processing, bioinformatic pipelines, and machine learning (ML), we extracted 21 features capturing copy number variations (CNVs) and fragmentomic characteristics to identify complementary signatures distinguishing OC from controls. Predictive models were developed using XGBoost with hyperparameter optimization and evaluated on an independent test set (n = 25% of the cohort). A dual-threshold classification strategy was applied to define an uncertainty zone and optimize screening performance.CNV-derived and fragmentomic features assessed in exploratory analysis on the training-validation set showed moderate discriminative power (AUC 0.569 - 0.946) but substantial overlap between groups. On the test set, the CNV-only model achieved an AUC of 0.855 (sensitivity 85%, specificity 50%), while the fragmentomics-only model reached an AUC of 0.8825 (sensitivity 95%, specificity 30%). Both feature domains captured complementary aspects of tumor-derived cfDNA, with fragmentomics favoring sensitivity and CNV-derived metrics improving specificity. Integration of both feature classes improved performance, yielding an AUC of 0.900, sensitivity of 85.00%, and specificity of 90.00%. SHAP analysis confirmed contributions from both feature types without a single dominant predictor.Conclusions: We present an integrative cfDNA framework for OC detection based on ulcWGS that combines CNV and fragmentomic signals to improve diagnostic performance over single-feature approaches. By enabling robust detection of tumor-associated patterns at ultra-low sequencing depth, this approach demonstrates that meaningful cancer discrimination can be achieved without reliance on deep sequencing. This highlights the potential of cost-effective and scalable liquid biopsy strategies for population-level cancer screening their integration into personalized and preventive oncology. Keywords: Liquid biopsy, ovarian cancer, ultra-low coverage whole genome sequencing, cell-free DNA, cell-free tumor DNA, cancer detection, copy number variations, insert size, fragmentomics, machine learning

Background Lymph node status and molecular subtype are among the most established prognostic factors in breast cancer. However, the extent to which their prognostic effects vary across different tumor size categories and clinical subgroups remains incompletely understood. We investigated the interplay between nodal status, molecular subtype, and tumor size in a large real world breast cancer cohort and developed a prognostic nomogram for individualized survival prediction. Methods A total of 12,225 women with invasive breast cancer from the Shiraz Breast Cancer Registry were analyzed. Patients were stratified according to tumor size, lymph node status, and molecular subtype. Overall survival (OS) and disease free survival (DFS) were evaluated using Kaplan Meier analyses and subgroup comparisons. Logistic regression was performed to identify predictors of lymph node involvement, while Cox regression was used to determine independent prognostic factors. A nomogram was subsequently developed and internally validated for prediction of 3-year and 5-year OS. Results Of 12,225 patients, 41.7% had lymph node positive disease. Across nearly all tumor size categories and molecular subtypes, nodal involvement was associated with significantly worse OS and DFS. Notably, the survival disadvantage associated with nodal positivity was more pronounced among patients with larger tumors and among those with HER2 positive and triple negative breast cancer (TNBC). Although TNBC demonstrated the lowest rate of lymph node involvement among molecular subtypes (adjusted OR 0.54, 95% CI 0.46-0.63), it appeared to show one of the largest survival gaps between node positive and node negative disease. In the overall cohort, survival outcomes generally ranked from best to worst as Luminal A, Luminal B, HER2 positive, and TNBC. However, survival differences among molecular subtypes were not consistently observed across all tumor size and nodal status subgroups. When significant differences were present, Luminal A and Luminal B tumors consistently showed superior outcomes compared with HER2 positive and TNBC tumors. Multivariable analysis identified lymph node status, tumor size, molecular subtype, lymphovascular invasion, tumor necrosis, type of surgery, radiotherapy, hormone therapy, and adjuvant chemotherapy as independent prognostic factors. A nomogram integrating clinicopathological and treatment variables demonstrated good predictive performance, with time dependent AUCs of 0.749 and 0.751 for 3 year and 5 year OS, respectively, and showed good calibration. Conclusions The prognostic impact of lymph node status is not uniform across breast cancer subgroups and appears particularly pronounced in larger tumors and biologically aggressive subtypes. Despite a lower likelihood of nodal involvement, TNBC showed substantial outcome deterioration when nodal metastasis was present. These findings highlight the importance of jointly considering nodal status, molecular subtype, and tumor burden in prognostic assessment.

Non-small cell lung cancer (NSCLC) is the most common form of lung cancer accounting for most cancer-related deaths worldwide. Despite substantial recent advances in targeted therapies and immunotherapy, the prognosis for advanced-stage disease remains comparably poor, which is why the identification of novel molecular biomarkers as well as therapeutic targets influencing tumor development, progression, and metastasis remain important. This study focusses on SERPINB13, a serine-protease inhibitor expressed in selected tissues that is dysregulated in several tumor entities. However, its role in NSCLC still remains largely unclear. We analyzed SERPINB13 transcription in a cohort of non-small cell lung cancer (NSCLC) cases including both lung squamous cell carcinoma (LUSC) and lung adenocarcinoma (LUAD) by transcriptome profiling. Epigenetic modifications were assessed via Methylation BeadChips. Additionally, SERPINB13 protein expression was assessed by immunohistochemistry (IHC) in an independent cohort of NSCLC comprising 126 LUSC patients. Correlation analyses were performed to associate SERPINB13 expression with key clinico-pathological parameters, including overall survival and extent of tumor-infiltrating immune cells. To functionally investigate the regulatory influence of peripheral blood mononuclear cells (PBMCs) on SERPINB13 expression in LUSC tumor cells in vitro, we utilized the SERPINB13-expressing LUSC cell line LUDLU-1. Here, gene transcription was analyzed by quantitative real-time PCR (RT-qPCR), confirmed by Western blot on the protein level. Transcriptome analysis revealed a significant upregulation of SERPINB13 in lung squamous cell carcinoma (LUSC) compared to lung adenocarcinoma (LUAD), highlighting a subtype-specific expression pattern. This differential expression was further associated with a distinct epigenetic DNA methylation signature at the SERPINB13 loci in LUSC, suggesting transcriptional regulation via hypomethylation. IHC analysis demonstrated that high SERPINB13 protein expression is significantly associated with prolonged overall survival in LUSC. Notably, SERPINB13 expression was enriched in immune-inflamed ("hot") tumors, characterized by elevated infiltrating lymphocytes and immune activation. Mechanistically, co-culture experiments with PBMCs induced SERPINB13 expression in a LUSC cell line in a dose- and time-dependent manner in the absence of direct cell contact. This suggests that soluble factors secreted by immune cells might play a key role in regulating SERPINB13 expression in the tumor microenvironment. Taken together, SERPINB13 is a novel prognostic indicator in LUSC that is modulated by Immune cells. Further studies are necessary to decipher the crosstalk of Immune cells on the Serpin B13 expressing tumor cells in depth, with regard to a possible interventional strategy. immunomodulatory potential strategies and personalized therapeutic approaches in NSCLC.

Objective To determine if it is possible to assess individual patient risk of the development of colorectal cancer (CRC) in people in high-risk groups due to their family history. Design/Method Retrospective observational study of prospectively collected data from consecutive patients referred for a colonoscopy. 2,478 consecutive patients were referred to a single colorectal surgical practice in Sydney, Australia between 1977 and 2018 for a colonoscopy because of a family history of CRC. Of these, 1,963 have been followed for more than 10 years and are the subject of this paper. Histopathological findings categorised as normal (N), non-advanced adenoma (NAA) or advanced neoplasia (AN) with AN proven to be the precursor to CRC. Intervention Colonoscopic screening on the basis of contemporary practice to 2006 and subsequently according to Australian National Health and Medical Research Council guidelines. Results Participants with normal or low-risk findings in the first decade remain at lower risk of CRC for 30 years from the commencement of screening. Conclusion It is possible to stratify individual patients in a high relative risk cohort into those with high or low personal risk of CRC based on colonoscopic findings in the first 10 years of surveillance. Those with no AN in the first ten years have a lower 30-year risk of developing AN than the general community. This offers the possibility of structuring surveillance programs around individual risk rather than group risk, lessening the need for multiple surveillance colonoscopies in the majority of such patients and improving the cost effectiveness of CRC screening at the population level.

Introduction. TAC/SARIFA has been introduced as a new robust and easy-to-evaluate biomarker in several cancer entities, including colorectal cancer. It is defined by direct contact between at least five tumour cells and one adipocyte and is believed to indicate metabolic reprogramming associated with adverse outcome. However, the mechanism that leads to TAC/SARIFA positivity remains unclear. To investigate whether there is an individual component, we conducted a study on double and triple cancers, establishing a within patient design. Methods. We retrospectively analysed a total of 135 cases with 276 colorectal cancers from two academic medical centres. The TAC/SARIFA status was evaluated, as were the basic histopathological factors. The median follow-up time was 120 months. Results. Cases with any TAC/SARIFA positive tumours showed significantly reduced overall survival (62 vs. 88 months; p = 0.011). Analysing the entire cohort, the rates of concordant and discordant cases followed a random distribution. However, restricting the analysis to synchronous pT3/4 cases revealed a significant deviation from a random distribution (p = 0.016). Conclusion. This study reveals significant concordance of TAC/SARIFA status in synchronous locally advanced colorectal double/triple carcinomas, supporting the concept that tumour adipocyte interaction reflects a host related microenvironmental condition linked to metabolic reprogramming rather than a purely tumour intrinsic event.

Colorectal cancer (CRC) is highly heterogeneous at both clinical and molecular levels, and the integration of circulating biomarkers with comprehensive genomic profiling remains limited. In this study, we measured circulating urokinase-type plasminogen activator (uPA) and its receptor (uPAR) in 53 patients with colorectal neoplasms and performed whole-genome sequencing (WGS) on matched tumor-normal pairs from 51 patients to characterize somatic mutations, copy number alterations (CNAs), tumor mutational burden (TMB), microsatellite instability (MSI), homologous recombination deficiency (HRD), and mutational signatures. Circulating uPAR levels were significantly elevated in patients with CRC compared with healthy controls, showing a stepwise increase across tumor stages and reaching the highest levels in stage IV disease. In contrast, circulating uPA levels showed only a non-significant trend toward elevation and did not vary significantly by stage. Despite the strong association between uPAR and tumor progression, circulating uPA and uPAR levels were not significantly correlated with TMB, MSI, HRD scores, or the mutational status of major CRC driver genes, including TP53, KRAS, FBXW7, BRAF, NRAS, and PIK3CA. Genomic analysis revealed a heterogeneous mutational landscape dominated by TP53 and APC, with only a minority of tumors exhibiting high TMB or MSI. Mutational signatures were primarily clock-like (SBS1, SBS5), with minimal contribution from MMR- or HRD-related processes. Together, these findings indicate that circulating uPAR is a robust marker of CRC progression that appears to operate largely independently of established genomic instability metrics. This supports uPAR potential utility in risk stratification and biological monitoring when integrated with molecular profiling.