Curcumin (CUM) can regulate the malignant behavior of glioblastoma (GBM). This study investigates whether CUM suppresses GBM progression by inducing cuproptosis via the AKT/Wnt/β-catenin signaling pathway. Optimal curcumin concentrations were determined using the MTT assay. A172 and U251 cells were treated with CUM, AKT inhibitor MK-2206, Wnt inhibitor LGK974, and Elsm-Cu (Elesclomol + CuCl2). Cell proliferation, migration, invasion, and apoptosis were assessed using carboxyfluorescein succinimidyl amino ester staining, scratch assays, Transwell assays, and flow cytometry, respectively. MitoSOX fluorescence, Seahorse metabolic analysis, immunofluorescence, Cu²⁺ detection, and Western blotting were used to evaluate mitochondrial oxidative stress and cuproptosis. The AKT/Wnt/β-catenin axis was analyzed using a TCF/LEF1 reporter kit and western blotting. A GBM xenograft model was established, and CUM was administered by gavage for five weeks. CUM effects on tumor growth, cuproptosis, and AKT/Wnt/β-catenin pathway protein expression were evaluated. Cells were treated with 10 µM and 20 µM CUM in vitro. CUM treatment reduced proliferation, migration, and invasion, while promoting oxidative stress and cuproptosis. CUM also suppressed Wnt/β-catenin signaling activity. Pathway inhibition increased reactive oxygen species (3.7-fold) and Cu²⁺ (3.1-fold) levels, and decreased dihydrolipoamide acetyltransferase expression, thereby restraining malignant behavior. In nude mice, CUM significantly reduced tumor growth, promoted cuproptosis, and inhibited AKT/Wnt/β-catenin axis activation. Our results indicate that CUM suppresses AKT/Wnt/β-catenin signaling, promotes cuproptosis, and interferes with GBM progression.

The approval of sotorasib and adagrasib as the first KRAS G12C inhibitors has made the RAS oncogene a druggable target. However, they have modest objective response rates and short response durations. Therefore, strategies for improving RAS-targeted cancer therapy are urgently needed. Here, we found that both sotorasib and adagrasib promoted topoisomerase IIα (Topo IIα) proteasomal degradation in KRAS G12C-mutant cancer cells and induced DNA damage and apoptosis. In cell lines with acquired resistance to sotorasib, elevated Topo IIα levels were detected. TOP2A overexpression in sensitive KRAS G12C-mutant cells conferred resistance to sotorasib, whereas TOP2A knockdown in sotorasib-resistant cell lines sensitized the cells to sotorasib. Moreover, the combination of a KRAS G12C inhibitor such as sotorasib with a Topo II inhibitor such as VP-16 synergistically decreased the survival of sotorasib-resistant RAS G12C-mutant cells with augmented induction of DNA damage and apoptosis, effectively inhibited the growth of sotorasib-resistant tumors, and delayed or prevented the emergence of acquired resistance to sotorasib in vivo. Collectively, our results reveal an essential role of Topo IIα inhibition in mediating the therapeutic efficacy of RAS-targeted cancer therapy, providing a strong scientific rationale for targeting Topo II to improve RAS-targeted cancer therapies.

Radiation therapy (RT) is the standard of care for glioblastoma but is not curative. Triggering the cGAS/stimulator of interferon genes (STING) pathway with potent agonists, such as 8803, exerts activity across high-grade glioma preclinical models. To determine if the combination of 8803 with RT warrants consideration in the up-front treatment setting and to clarify the underlying mechanisms of therapeutic activity, C57BL/6J mice harboring intracerebral CT-2A or QPP8v gliomas were treated with RT, intratumoral 8803, or both. The treatment with the combination resulted in 80% long-term survival in the CT-2A model but not in the radiation-resistant QPP8v model. This therapeutic effect was maintained in Sting-/- CT-2A cells, highlighting the direct role of the immune system in mediating the survival benefit. Single-cell RNA-Seq identified increased nitric oxide synthase 2 (Nos2) in inflammatory tumor-associated macrophages; however, the therapeutic effect was maintained in Nos2-/- mice. Additionally, 8803 reprogrammed the blood-brain barrier (BBB) by altering the Pecam and Cd147 pathways in endothelial cells; intracranial injection of 8803 induced bihemispheric BBB opening for up to 24 hours. Sting activation was visualized longitudinally using 3'-deoxy-3'-[18F]-fluorothymidine ([18F]-FLT) PET, which peaked 72-96 hours after 8803 administration. In summary, 8803 combined with RT triggers distinctive antiglioma immune reactivity, facilitates BBB opening, and warrants consideration for up-front clinical trials in glioblastoma, where treatment effects can be monitored using [18F]-FLT PET imaging.

The hypoxic microenvironment is a critical feature of malignant progression in non-small cell lung cancer (NSCLC). However, the regulatory role of circular RNAs (circRNAs) in this context remains incompletely understood. Hypoxia-related circRNAs were screened by integrating GEO datasets, and the expression of circ_0017521 in hypoxia-treated cells was validated using qRT-PCR. RNase R digestion and actinomycin D assays were employed to assess its stability. Biological functions were evaluated through CCK-8, colony formation, Transwell, flow cytometry, and glycolytic metabolism assays. Dual-luciferase reporter, RNA pull-down, and rescue experiments were conducted to elucidate the circ_0017521/miR-532-3p/PFKFB3 axis. A nude mouse xenograft model was constructed. circ_0017521 was specifically upregulated in NSCLC tissues and hypoxia-treated cells, exhibiting remarkable stability compared to linear AKR1E2 and primarily localizing in the cytoplasm. Under hypoxic conditions, knockdown of circ_0017521 noticeably inhibited NSCLC cell proliferation, migration, invasion, epithelial-mesenchymal transition (EMT), and glycolysis, while promoting apoptosis. Mechanistically, circ_0017521 competitively bound to miR-532-3p, relieving its suppression of PFKFB3 and thereby activating the PI3K/AKT signaling pathway. Upregulation of PFKFB3 drove the expression of glycolytic enzymes (GLUT1, LDHA, HK2) and synergistically activated EMT. Animal experiments confirmed that silencing circ_0017521 suppressed tumor growth and glycolysis. This study revealed that hypoxia-induced circ_0017521 activated the PI3K/AKT pathway through the miR-532-3p/PFKFB3 axis, synergistically driving EMT and glycolysis, thereby promoting NSCLC progression.

Triple-negative breast cancer (TNBC) is an aggressive disease characterized by high metastatic potential and limited treatment options. Protein kinase C-eta (PKCη), an antiapoptotic kinase of the novel PKC subfamily, is associated with poor prognosis in breast cancer patients. Analysis of TNBC tumors revealed that PRKCH (PKCη) expression is linked to an epithelial‒mesenchymal transition (EMT) signature, which is indicative of a metastatic phenotype. Using genetic ablation studies, we showed that PKCη promotes metastasis by enhancing EMT and stemness. Notably, compared with those in PKCη-intact tumors, orthotopic xenografts of PKCη-knockout cells in NSG mice resulted in reduced tumor growth and metastasis. Mechanistically, PKCη functions as a negative regulator of the Hippo pathway by activating YAP. PKCη phosphorylates YAP at Ser128, leading to its stabilization and nuclear translocation, which promotes metastasis. We also demonstrated that PKCη negatively regulates AKT, thereby further sustaining the downregulation of the Hippo pathway. Finally, we show that an evolutionarily conserved peptide encoded by an upstream open reading frame (uORF) preceding the PKCη coding sequence functions as a PKCη degrader, activating the Hippo pathway and promoting YAP degradation. Together, our findings reveal a PKCη-driven signaling axis that regulates the Hippo-YAP pathway in TNBC metastasis, highlighting the potential therapeutic vulnerability of this aggressive disease.

Ferroptosis plays a critical role in non-small cell lung cancer (NSCLC) progression. Curculigoside (Cur) exhibits anti-cancer properties. This study aimed to elucidate the precise action of Cur on NSCLC ferroptosis. Various concentrations of Cur were used to treat A549 and H520 cells to evaluate its cytotoxicity. The regulation of Wilms' tumor 1-associating protein (WTAP) on GTP cyclohydrolase 1 (GCH1) was verified by methylated RNA immunoprecipitation (MeRIP) and mRNA stability assays. Cur suppressed proliferative, migratory, and invasive capacities of A549 and H520 cells in vitro and reduced tumor growth in A549-derived subcutaneous xenografts in vivo. Cur induced ferroptosis in A549 and H520 cells. Mechanistically, Cur decreased the protein levels of GCH1 and WTAP, and WTAP mediated the N6-methyladenosine (m6A) methylation and stabilization of GCH1 mRNA. GCH1 depletion promoted NSCLC cell ferroptosis and impeded their malignant phenotypes. Moreover, overexpression of GCH1 reversed Cur-induced ferroptosis and malignant phenotype inhibition in A549 and H520 NSCLC cells. Our study suggested that Cur induced ferroptosis and suppressed malignant phenotypes in NSCLC in part through the WTAP-GCH1 axis, thereby revealing a novel mechanism for its therapeutic potential.

This study aimed to describe the implementation and functioning of a regional hub-and-spoke model ("Gaslini Diffuso") for managing pediatric bronchiolitis in Liguria, Italy, during the 2023-2024 season, focusing on severity stratification, resource allocation, and outcomes. A retrospective observational study was conducted across one tertiary hub (IRCCS Istituto Giannina Gaslini, Genoa) and four affiliated spoke hospitals. Medical records of all patients aged 0-2 years hospitalized with bronchiolitis (ICD-9-CM 466.19) between October 2023 and March 2024 were reviewed. Demographic, clinical, microbiological, and treatment data were analyzed. Predictors of centralization to the hub were identified through multivariable logistic regression. A total of 562 patients were included (median age 95 days; 40.4% female). Most cases were mild to moderate, with 56.6% requiring respiratory support-mainly low-flow oxygen or HFNC-and only 2% requiring mechanical ventilation. Thirteen patients (2.3%) were admitted to the PICU, and no deaths occurred. Centralized patients (n = 10) were significantly younger (median 43.5 days) and had higher severity indicators, including elevated CO₂ and CRP levels, and longer respiratory support (median 5 vs. 3-4 days, p < 0.001). Independent risk factors for centralization were age < 60 days (OR 23.1, p = 0.004) and HFNC use (OR 20.5, p = 0.006). Spoke centers showed homogeneous adherence to referral criteria, though some variability in ancillary treatments persisted.

 The Ligurian hub-and-spoke model demonstrated internal consistency between referral criteria and observed patient severity, supporting appropriate case stratification within the regional network. This integrated framework enhanced regional coordination and represents a scalable, sustainable model for pediatric respiratory disease management.

• Bronchiolitis is the leading cause of hospitalization in infants under two, with seasonal surges that may overwhelm pediatric services; management remains largely supportive. • Hub-and-spoke models have been proposed to optimize care and resource allocation, but real-world data on their clinical and organizational impact, especially post-COVID, is limited.

• This study evaluates, for the first time in Italy, the real-world implementation of a regional hub-and-spoke model (Gaslini Diffuso) for bronchiolitis management. • The model enabled effective stratification of disease severity, with high specificity in centralizing only the most critical cases, ensuring efficient use of pediatric intensive care resources.

Postoperative respiratory events are a common complication of tonsillectomy for pediatric obstructive sleep apnea (OSA). Despite identified risk factors, there are no validated criteria for systematic postoperative intensive care unit (ICU) admission. In France, an apnea-hypopnea index (AHI) ≥ 10 events/h is a recommended criterion. This study aimed to assess the incidence and risk factors of early postoperative respiratory complications in children undergoing tonsillectomy for OSA with planned ICU admission. We conducted an observational study of children undergoing tonsillectomy between January 2020 and December 2022, with a preoperative ICU indication. Respiratory events were defined as a need of oxygen therapy, noninvasive ventilation (NIV), or tracheal intubation. Collected data included anthropometrics, comorbidities, AHI, gas exchange parameters, and surgical details. Univariate and multivariate analyses were performed for potential risk factors associated with postoperative respiratory complications. 166 children were included (median age 3 years [IQR 2-5]). The median preoperative AHI was 25 events/hour [13-43]. Overall, 159 (96%) patients had at least one comorbidity (45 Down syndrome, 31 obesity and 25 craniofacial malformations). Thirty-four (21%) patients had a respiratory complication: 30 needed oxygen therapy, 4 required NIV. None required an intubation. Age (p < 0.001), percentage of sleep time spent with an Oxygen saturation level < 85% (T85) and < 90% (T90) (p = 0.02 and p = 0.03, respectively) and McGill Oximetry Score (p = 0.01) were significantly associated with respiratory events, whereas neither AHI nor comorbidities showed a significant association. Conclusion: This study highlights the limited value of AHI alone in predicting respiratory complications post-tonsillectomy.

Cystic fibrosis (CF), a genetic disorder stemming from CFTR gene mutations, requires accurate risk prediction to improve management. Modulator therapies have advanced treatment but remain limited, as they don't cover all gene variants and face accessibility issues. To address these challenges, a novel Cystic Fibrosis Risk Prediction Framework (CGRPF) is proposed. CGRPF utilizes mean imputation for missing data, the Fox Wolf Optimizer (FWO) for effective feature selection, and an AttentiveLSTM to capture temporal patterns in time-series data, aiding chronic disease prediction. Fully connected layers and a softmax layer enhance model performance and ensure calibrated classification into high, medium, and low-risk categories. Tested on the CFTR-2 dataset, CGRPF achieved strong performance metrics - 97 % accuracy, 91 % precision, 97 % recall, 93 % F1-score, outperforming state-of-the-art models in CF risk prediction.

Cannabis-based medicine may alleviate breathlessness. This study will investigate whether dronabinol, a synthetic form of Δ⁹-tetrahydrocannabinol (Δ⁹-THC), reduces breathlessness in patients with severe and very severe chronic obstructive pulmonary disease (sCOPD) compared to placebo.

This single-centre, randomised, double-blinded, placebo-controlled, crossover trial will enrol 30 patients with sCOPD and persistent breathlessness despite optimal treatment. Patients will be recruited from a pulmonary outpatient clinic in Denmark over 24 months. Eligible patients (aged ≥18 years) will receive either dronabinol or placebo for 4 weeks, followed by a 2-week washout, before crossing over to the other treatment for 4 weeks. Exclusion criteria include ongoing infection, substance abuse and significant comorbidities. Primary outcome is breathing discomfort or unpleasantness measured using the 0-10 Numerical Rating Scale. Secondary outcomes include lung function (forced expiratory volume in one second), hair cortisol concentrations, functional tests, plasma THC blood concentrations and questionnaires assessing breathlessness, activity, quality of life, anxiety and depression. Continuous monitoring of vital signs, activity and sleep will be performed using a Garmin Venu 3 smartwatch. Data will be entered into electronic case report forms and monitored by the Good Clinical Practice (GCP) unit in Odense.

This will be the largest randomised, double-blinded, crossover trial to investigate dronabinol in patients with COPD and will provide new knowledge on the efficacy and safety.

Written informed consents will be obtained from study patients. The study has been approved by the Danish Medicines Agency (case number: 2023010659) and the medical research ethics committees (case number: 2301456). It is registered in the European Union Clinical Trials Registry (2024-513593-22-00) and ClinicalTrials.gov (NCT06473701). The trial follows the Declaration of Helsinki II and International Council for Harmonisation-GCP guidelines. Findings will be disseminated in peer-reviewed publications.

The European Union Clinical Trials Registry (2024-513593-22-00) and ClinicalTrials.gov (NCT06473701).