Head and neck squamous cell carcinoma (HNSC) remains one of the main causes of cancer-related deaths among male patients with cancer. Although surgery, chemotherapy, radiotherapy, targeted therapy, and immunotherapy are available, their success in controlling the disease remains limited. Novel targets and therapeutics for HNSC therefore await identification, which is frequently pursued using bioinformatics approaches.

Bioinformatics analyses were conducted to evaluate prognosis, differential gene expression, pathways, therapeutics, immunohistochemistry, and expression correlations. Experimental procedures included gene knockdown, proliferation assays, cell cycle analysis, drug treatment, 3D assays, proximity ligation assays, immunoprecipitation, and immunohistochemistry.

In the present study, a prognostic factor for HNSC with concordant expression changes was identified. Ring finger protein 149 (RNF149) was shown to predict poor prognosis and to be upregulated during tumorigenesis. Pathway and therapeutic analyses revealed that RNF149-high HNSCs were enriched for cell cycle dysregulation, which might be counteracted by cyclin-dependent kinase inhibitors (CDKi). Cell line experiments validated that RNF149 knockdown in HNSC decreased proliferation and altered the response to CDKi in both 2D and 3D environments. Multi-omics analysis and tissue staining revealed that this ubiquitin ligase might ubiquitinate cyclin-dependent kinase inhibitor 2C (CDKN2C) in HNSC. This regulation was experimentally verified using proximity ligation assays (PLA) and immunoprecipitation. Finally, tissue array staining confirmed a negative correlation between RNF149 and CDKN2C expression in HNSC.

This study provides insights into RNF149-mediated proteolysis in HNSC.

Gastric cancer (GC) remains a leading cause of cancer-associated deaths globally, particularly in East Asia. Although anti-angiogenic therapies have yielded therapeutic benefit in GC, their efficacy is limited, as current vascular endothelial growth factor-A (VEGF-A) and VEGF receptor-2 (VEGFR-2) targeted therapies eventually fail due to compensatory pathways involving VEGF-B and placental growth factor (PlGF). PB101, a VEGFR-1 decoy receptor, inhibits VEGF-A, VEGF-B, and PlGF, potentially offering broader anti-angiogenic effects. This study evaluated the efficacy of PB101 alone and in combination with cytotoxic chemotherapeutic agents in GC models.

In vitro assays including CellTiter-Glo cell viability, tube formation, transwell and chemotaxis migration were conducted to evaluate PB101's effects on cell survival, migration, and endothelial angiogenesis. An in vivo NCI-N87 xenograft model was used to evaluate antitumor efficacy. Tumor angiogenesis was assessed by CD31 immunohistochemistry.

PB101 exerted no direct cytotoxicity on GC cells, either alone or in combination with chemotherapy. However, it demonstrated potent anti-angiogenic properties by significantly inhibiting endothelial cell migration and tube formation. As 2D culture cannot recapitulate the tumor microenvironment (TME), in vivo studies were conducted. In vivo studies revealed that mice receiving PB101 plus paclitaxel or irinotecan exhibited greater tumor volume suppression compared to each single-agent treatment group. Harvested tumors from PB101 and combination groups showed reduced CD31 expression, indicating reduced angiogenesis.

PB101, blocking VEGF-A/B and PlGF, showed broad anti-angiogenic activity and enhanced chemotherapy efficacy in GC xenograft models when used in combination.

Colorectal cancer (CRC) remains the leading cause of cancer-related mortality worldwide and necessitates the development of novel therapeutic strategies. The tumor immune microenvironment (TME) critically influences disease progression and the response to immune checkpoint inhibitors (ICIs). Tumor-infiltrating lymphocytes (TILs) are key components of the TME with established prognostic and predictive significance. Nevertheless, detailed TIL characterization using flow cytometry has not been fully investigated in CRC.

We analyzed TILs from 90 fresh CRC specimens using multicolor flow cytometry to investigate the association between specific T cell subsets and clinical outcomes. Patients were classified into Hot and Cold groups based on hierarchical clustering of TIL marker expression.

The Hot group demonstrated significantly better overall survival (OS) compared to the Cold group (5-year OS: 86.7% vs. 63.9%, p=0.006), although recurrence-free survival (RFS) was not significantly different (5-year RFS: 79.5% vs. 66.1%, p=0.24). CITRUS analysis revealed that PD-1⁺Tim-3⁺CD103⁺CD8⁺ T cells were enriched in hot tumors (32.1% vs. 6.1%, p<0.001) and correlated with a favorable prognosis. Importantly, multivariate analysis demonstrated that a low frequency of PD-1⁺Tim-3⁺CD103⁺ cells among CD8⁺ T cells was an independent prognostic factor for OS [hazard ratio (HR)=3.36, 95% confidence interval (CI)=1.20-9.34, p=0.02].

A high frequency of PD-1⁺Tim-3⁺CD103⁺ CD8⁺ T cells is associated with better survival in CRC, highlighting their potential as a prognostic biomarker and therapeutic target.

Replication Protein A (RPA) is a key single-stranded DNA (ssDNA)-binding protein essential for maintaining genome integrity during DNA replication, repair, and recombination. In this study, we elucidate the mechanisms by which a small-molecule RPA inhibitor induces functional RPA exhaustion. Using non-small cell lung cancer and BRCA1-deficient breast and ovarian cancer models, we demonstrate that RPA is critical for sustaining replication fork speed under normal conditions and for facilitating replication restart following fork stalling. Disruption of replication fork-associated processes, including Okazaki fragment processing and ssDNA gap suppression, increases cellular dependence on RPA for ssDNA protection. Chemical inhibition of RPA exacerbates genome instability in BRCA1-deficient cancer models treated with PARP inhibitors, leading to loss of ssDNA gap protection, chromosome shattering, and ultimately, cell death. Combining genetic and pharmacologic approaches to induce ssDNA accumulation alongside RPA exhaustion in vivo shows therapeutic efficacy in BRCA1-deficient breast cancer. These findings provide a mechanistic framework for targeting RPA-mediated ssDNA protection as a therapeutic strategy in cancers experiencing endogenous or therapy-induced replication stress.

We present a case of recurrent thymic neuroendocrine neoplasm (NEN) in a 43-year-old male, who developed ectopic Cushing syndrome (CS), 8 years after his initial diagnosis. Due to persistent biochemical hypercortisolism and clinical symptoms of CS, he was trialled on multiple adrenal steroidogenesis inhibitors with surgical resection and radiotherapy. Eventually, he was transitioned to osilodrostat, a new potent adrenal steroidogenesis inhibitor. Following multidisciplinary discussion, he underwent peptide receptor radionuclide therapy (PRRT). After 12 months of therapy, his cortisol levels have normalised. This is a novel case of functional thymic NEN that was biochemically responsive to osilodrostat and PRRT. Osilodrostat predominantly blocks 11-β hydroxylase leading to a reduction in biochemical hypercortisolism and improvement in clinical manifestations. Initial evidence supported its role in the setting of Cushing's disease. There is growing evidence for its use in the setting of ectopic CS as observed with our case. PRRT continues to have a role in the setting of functional NENs.

Ectopic CS is a rare, functional syndrome most often seen with neuroendocrine tumours. Management relies on treatment of the primary pathology as well as hypercortisolism, for which there are new novel agents available including osilodrostat. Directed therapies such as peptide receptor radionuclide therapy can have favourable outcomes in addressing metabolic and biochemical disease in the setting of ectopic CS.

Borderline ovarian tumors (BOT) and low-grade serous carcinoma (LGSC) are rare epithelial ovarian neoplasms with distinct clinicopathologic features and generally more favorable outcomes than high-grade epithelial tumors. This study aimed to describe the clinical, pathologic, and treatment characteristics of BOT and LGSC treated in Argentine and Uruguayan oncology centers, and to report oncologic outcomes.

A multicenter retrospective study was conducted including 369 adult patients diagnosed between 2009 and 2020 in 19 hospitals in Argentina and 1 in Uruguay. Clinical, pathologic, surgical, treatment, and survival data were collected from medical records.

Of the 369 patients, 336 (91.1%) had BOT and 33 (8.9%) had LGSC. Most tumors were diagnosed at an early stage. BOT was more frequently associated with stage I disease, conservative surgery, and very limited use of systemic treatment, whereas LGSC showed more bilateral disease, ascites, neovascularization, invasive pathologic features, greater use of staging-related procedures, and more frequent systemic treatment. In BOT, 5-year overall survival (OS) and recurrence-free survival (RFS) were 98.9% and 90.2%, respectively. In LGSC, the corresponding rates were 91.4% and 75.5%.

BOT and LGSC showed clearly different clinical, pathologic, treatment, and survival profiles in this South American multicenter cohort. BOT was usually diagnosed at an early stage and had excellent oncologic outcomes, whereas LGSC was associated with more aggressive clinicopathologic features, more frequent systemic treatment, and poorer survival.

Colorectal cancer (CRC) is a leading cause of cancer-related deaths worldwide. Although epigenetic alterations are common in CRC, the epigenetic changes that occur during colorectal carcinogenesis remain unclear. Thus, we sought to elucidate the role of NOS1 methylation in colorectal carcinogenesis, which is essential for understanding disease pathology. We used the UCSC Xena database to comparatively analyze NOS1 expression and methylation status between tumour and adjacent normal tissues across 33 cancer types. Low expression and hypermethylation of NOS1 have been identified in 12 cancer types, with CRC demonstrating this characteristic epigenetic regulation. NOS1 promoter methylation status was examined using genomic DNA extraction and MassARRAY EpiTYPER methylation analysis. NOS1 hypermethylation was confirmed among CRCs in in-house dataset 1 (p < 0.001), and among CRCs and advanced adenomas in in-house dataset 2 (p < 0.001). An upward trend in methylation changes was identified from non-advanced adenoma to advanced adenoma to CRC (p for trend < 0.001). Quantitative real-time PCR was used to analyze NOS1 expression in in-house Dataset 3, and significantly low NOS1 expression was also identified in CRCs (p < 0.001). Kaplan-Meier estimator and Cox proportional hazard models were used to assess the prognostic and predictive roles of NOS1. NOS1 hypermethylation was significantly associated with better disease-specific survival (DSS) in CRC patients. NOS1 hypermethylation is an epigenetic driver of colorectal tumorigenesis and is associated with better survival.

DNA methylation, is catalyzed by DNA methyltransferases (DNMTs), and its aberrant patterns are implicated in thyroid cancer pathogenesis. The study aimed to investigate the association of DNMTs with thyroid cancer and evaluated the effects of sustained demethylating therapy in a cell-based study. DNMTs expressions in thyroid cancer were analyzed using GEO and TCGA datasets. Additionally, 16 paired and three unpaired papillary thyroid carcinoma (PTC) samples from Taipei Medical University (TMU), along with commercial tissue arrays, were analyzed. Furthermore, the effects of the covalent DNMT inhibitor, 5-azacytidine (5-Aza), and the DNMT1-selective inhibitor, GSK-3484862, on cell viability were evaluated in PTC and follicular thyroid carcinoma (FTC) cell lines. DNMT1 and DNMT3A were upregulated in PTC, with DNMT1 expression correlated with the BRAF mutation and lymph node invasion in TCGA data, findings further confirmed in the TMU cohort and tissue arrays. Short-term (24 h) 5-Aza treatment (1 and 5 µM) induced substantial cell death regardless of the DNA methylation status, whereas short-term GSK-3484862 (5 µM) treatment showed minimal cytotoxicity. In contrast, sustained low-dose GSK-3484862 treatment (approximately 1-3 weeks at 2 µM) effectively reduced global DNA methylation and decreased cell viability of TPC-1 and FTC-236 cells through apoptosis, rather than by inhibiting proliferation. In conclusion, DNMT1 overexpression in PTC suggests its involvement in thyroid carcinogenesis. Sustained inhibition of DNMT1 effectively reduced global DNA methylation and promoted apoptosis, highlighting the potential of prolonged DNMT1-targeted therapy. Further in vitro and in vivo studies are warranted to validate these results and elucidate the underlying mechanisms.

The role of lifestyle interventions in treatment success has become essential for nearly every disease. Healthy dietary habits, regular exercise, and stress management are key pillars that can improve quality of life during treatment, as well as delay disease onset and progression. In this study, we focus on the combination of mild calorie restriction (CR) and voluntary exercise as coadjuvants to chemotherapy in the treatment of triple-negative breast cancer using the 4T1 mouse model. In this model, voluntary exercise did not add benefits beyond chemotherapy plus CR in terms of primary tumor size, body composition, or physical performance, while dampening the antimetastatic effect of CR in the lungs of sedentary mice. These findings highlight the challenges of translating results from one preclinical model to another, and ultimately to humans. Diet and exercise type and chemotherapeutic agent influence the outcomes of each type of cancer. More research is needed to determine if and when exercise should be recommended to particular cancer patients.

In surgical oncology, precise intraoperative delineation of malignant tumors for complete yet non-excessive resection has been challenging. Near-infrared II (NIR-II, 1,000 to 3,000 nm) imaging enables deep-tissue, high-contrast visualization, but clinical translation is limited by the lack of robust probes. Cytalux, the first clinically approved NIR-I (700 to 1,000 nm) probe for folate receptor (FR)-positive cancers, is limited in shallow imaging depth and low tumor to normal tissue contrast. Here, we present "Octopus" (OCTP), a modular multi-arm PEG-based NIR-II probe targeting FR capable of rapid and sustained tumor accumulation upon systemic administration to afford superior tumor-to-background ratios and precise margin visualization by NIR-II imaging, significantly outperforming Cytalux in mouse models. Importantly, we uncovered that NIR-II imaging in the >1,300 nm emission range under NIR-I excitation completely suppressed tissue autofluorescence, allowing unambiguous molecular imaging of residual cancerous cells at tumor margins for resection. Pharmacokinetic and toxicity studies demonstrate rapid clearance, minimal off-target accumulation, and excellent biocompatibility. OCTP enables accurate, background-free >1,300 nm fluorescence-guided tumor surgery, promising as the next-generation NIR-II molecular imaging of cancer.