Head and neck squamous cell carcinoma (HNSCC) remains a leading cause of cancer mortality worldwide, with treatment resistance posing a major clinical challenge. The tumor microenvironment, particularly the three-dimensional (3D) extracellular matrix architecture, plays a critical role in driving therapy resistance, yet the underlying molecular mechanisms remain incompletely understood. Here we investigate how the 3D collagen microenvironment orchestrates chemoresistance through NF-κB/STAT3 signaling and partial epithelial-mesenchymal transition (EMT) in HNSCC. Using 3D type I collagen culture systems, both FaDu and CAL27 HNSCC cells form spheroidal structures with cytoskeletal reorganization. Compared to 2D monolayers, 3D collagen culture activated NF-κB signaling, upregulated stemness marker ALDH1A1 and drug resistance proteins ABCG2/Bcl2, and significantly increased IC50 values for paclitaxel and cisplatin in FaDu and CAL27 cells. NF-κB inhibitor PDTC reversed these phenotypes, reducing resistance marker expression and restoring chemosensitivity. In FaDu xenografts, PDTC combined with paclitaxel exerted enhanced antitumor effects, achieving greatest tumor growth inhibition. Mechanistically, we identified a signaling hierarchy with NF-κB upstream of STAT3; PDTC suppressed both pathways, while Stat3 inhibitor Stattic only affected STAT3 downstream effectors. Clinical relevance was confirmed by immunohistochemistry of human HNSCC specimens, showing spatial enrichment of p65/p-p65 at the tumor-stroma interface specifically in lymph node-positive cases, with p-p65 H-scores significantly higher in metastatic patients (p < 0.05). Importantly, 3D collagen induced a partial EMT state with concurrent E-cadherin and N-cadherin upregulation, downregulation of ZO-1 and upregulation of Slug. Using a 14-gene p-EMT score in chemoresistant patient data, RELA and STAT3 strongly correlated with this hybrid score (RELA: R = 0.75; STAT3: R = 0.82) and activated mesenchymal genes while preserving epithelial identity. These findings demonstrate that the 3D collagen microenvironment activates an NF-κB-Stat3 signaling axis that drives cancer stemness, p-EMT, and chemoresistance in HNSCC, highlighting NF-κB inhibition as a promising therapeutic strategy to overcome microenvironment-mediated resistance.

Emergency palliative radiotherapy is often delivered under time-constrained and high-stress conditions, frequently bypassing conventional computed tomography (CT) simulation and relying on nonstandard workflows. These factors increase variability and the potential for error.

This work describes the development, implementation, and early clinical evaluation of a simulation-free, cone-beam CT (CBCT) based emergency radiotherapy workflow that leverages treatment planning system (TPS) scripting to standardize and automate deterministic steps while preserving clinical oversight.

An ESAPI scripting-driven emergency workflow was developed within the TPS to automate prescription handling, plan generation, beam configuration, dose calculation, and setup preparation using CBCT images. Safety checks were implemented in the form of prompts with the appropriate actions required. Alternatively, manual input was retained for image verification and treatment field borders definition. The workflow was implemented across multiple clinical sites within a single healthcare system and evaluated through dosimetric comparison of CT- and CBCT-based planning. Metric parameters such as workflow time, reductions in manual interactions, and pre- and post-training survey outcomes of automated 3D workflow were compared to the same parameters of the conventional 2D workflow.

The automated workflow successfully generated clinically acceptable emergency treatment plans with minimal manual input. CBCT-based dose calculations demonstrated agreement with CT-based planning within 2.5% when heterogeneity corrections were enabled and within 4.7% when disabled. Automation eliminated 33 manual data entry steps present in the 2D workflow and enabled treatment delivery with average on-table time of approximately an hour. Survey results demonstrated strong staff support for automation, with over 90% anticipating improvement prior to training and 100% reporting perceived improvement following training. Staff comfort and confidence improved post-training, and concerns regarding workflow clarity and safety decreased.

A scripting-based, simulation-free emergency radiotherapy workflow was developed for safe and standardized palliative treatments while maintaining established clinical safeguards. By automating deterministic planning tasks and preserving human oversight for clinical decision-making, the proposed approach improves consistency and staff confidence and has potential to improve safety and accuracy of palliative treatments.

NPC2 is a lysosomal cholesterol transporter involved in intracellular lipid trafficking and cholesterol homeostasis, processes that have been increasingly implicated in cancer biology. To quantify the clinical impact of NPC2 expression across human malignancies, we analyzed survival data from 2,097 patients evaluated for NPC2 expression in tumor tissues. We conducted a random-effects meta-analysis of four eligible studies assessing NPC2 expression by mRNA (RNA-seq, qRT-PCR) and protein-based methods (immunohistochemistry, Western blot).The cohort included 756 patients with gastric cancer (36.0%), 374 with hepatocellular carcinoma (17.8%), and 967 with gliomas (46.2%). Pooled analyses demonstrated that elevated NPC2 expression was significantly associated with worse overall survival across cancers (HR 1.29, 95% CI 1.10-1.52, p = 0.001; I² = 56%). In gastric cancer, this association was stronger and highly consistent (HR 1.37, 95% CI 1.18-1.60, p < 0.001; I² = 0%). These results suggest that NPC2 overexpression identifies a subset of tumors with increased aggressiveness and adverse clinical outcomes. Collectively, these findings support NPC2 as a candidate adverse prognostic biomarker and highlight lysosomal cholesterol trafficking as a pathway of translational interest in cancer biology. Further mechanistic and clinical studies are warranted to clarify the biological role of NPC2 and its potential therapeutic implications.

ObjectiveLung cancer is the leading cause of cancer-related mortality worldwide, with lung adenocarcinoma representing the predominant subtype. The increasing incidence of lung adenocarcinoma underscores the urgent need to establish novel therapeutic targets for clinical intervention.MethodsWe performed Mendelian randomization and colocalization analyses on large-scale lung adenocarcinoma genome-wide association study datasets. Bulk transcriptome and single-cell RNA sequencing were further applied to explore the function of candidate targets and related cell types.ResultsMendelian randomization revealed that elevated RNASET2 expression causally reduced the risk of lung adenocarcinoma (odds ratio = 0.817, 95% confidence interval: 0.754-0.885, p < 0.001), whereas increased BTN3A2 expression was associated with an elevated disease risk. Colocalization analysis supported a shared causal variant between RNASET2 and lung adenocarcinoma (posterior probability of hypothesis 4 = 78.6%). Bulk transcriptome data demonstrated that RNASET2 was upregulated in tumor tissues and was associated with better prognosis. Single-cell RNA sequencing revealed that macrophages were the predominant cell type expressing RNASET2, whereas weak expression was also detected in pulmonary epithelial cells and several other cell populations. There was a trend toward reduced abundance of high RNASET2-expressing macrophage subcluster 0 in tumor tissues, whereas low RNASET2-expressing subcluster 8 tended to accumulate within tumor lesions. Enrichment analysis of their signature genes suggested opposing biological roles in lung adenocarcinoma progression, which warrants further functional verification.ConclusionsRNASET2 is closely associated with the occurrence and prognosis of lung adenocarcinoma. Macrophage-expressed RNASET2 represent a promising candidate molecule for further therapeutic exploration.

Vascular endothelial growth factor receptor 2 (VEGFR-2) is a major mediator of pathological angiogenesis, which is critical to tumor progression and metastasis; therefore, inhibition of VEGFR-2 remains a key therapeutic strategy for cancer. Despite the established clinical efficacy of several approved VEGFR-2-targeted agents, many first-generation inhibitors are limited by dose-limiting toxicities and the emergence of resistance, highlighting an ongoing need for next-generation compounds with improved selectivity and more favorable safety profiles. Among nitrogen-containing heterocyclic frameworks, triazoles and imidazoles are widely employed in medicinal chemistry owing to their favorable structural features, including appropriate stability, synthetic accessibility, and the ability to engage in key molecular interactions. This review summarizes recent advances in the design of triazole- and imidazole-based VEGFR-2 inhibitors, containing Schiff bases, bicyclic, tricyclic, and glycoside derivatives. We comparatively describe their VEGFR-2 inhibitory activities and anticancer cytotoxicity data, and discuss key structure-activity relationships and mechanistic insights, with particular emphasis on molecular features that appear to favor potent and selective VEGFR-2 inhibition. This review compiles in vitro, in vivo, and in silico data on triazole- and imidazole-based VEGFR-2 inhibitors, providing a basis for understanding trends relevant to the design and development of optimized anti-angiogenic agents.

N4-acetylcytidine (ac4C) RNA modification has emerged as a critical epigenetic regulator in tumorigenesis and progression. However, the comprehensive landscape of ac4C modification in colorectal cancer (CRC), particularly its orchestration of the tumor microenvironment (TME) and stromal crosstalk, remains largely unexplored.

This study aimed to comprehensively delineate the landscape of N4-acetylcytidine (ac4C) RNA modification in colorectal cancer (CRC) to uncover its role in shaping the tumor microenvironment and clinical outcomes. Integrated bioinformatic and in vitro analyses revealed that ac4C patterns dictate aggressive stromal crosstalk and identified CERCAM as a cancer-associated fibroblast (CAF)-specific driver of tumor progression, providing a novel framework for prognostic prediction and personalized targeted therapy.

Transcriptomic and clinical data of CRC patients were curated from TCGA and GEO databases (GSE14333, GSE17536, GSE38832, GSE39582). The ac4C modification patterns were quantified using single-sample Gene Set Enrichment Analysis (ssGSEA). Single-cell RNA sequencing (scRNA-seq) analyses were performed via the scCancer Explorer database. A prognostic ac4C score was constructed to evaluate clinical outcomes and therapeutic vulnerabilities. The functional role of the identified target gene, CERCAM, was validated using in vitro co-culture experiments.

Patients with high ac4C scores exhibited worse prognosis, higher clinical stages, and metastasis (N2, M1). High ac4C levels positively correlated with epithelial-mesenchymal transition (EMT), TGF-β signaling, and extracellular matrix (ECM) receptor interactions, whereas low ac4C levels were associated with cell cycle, fatty acid metabolism, and microsatellite stability (MSS). The TME of high-ac4C tumors was characterized by higher Stromal, Immune, and ESTIMATE scores, elevated immune checkpoints (LAG3, CD14, LILRB2, SIRPA, CD8A), and strong correlations with macrophages and NK cells. scRNA-seq analysis revealed that CERCAM, a key gene within the ac4C network, was predominantly expressed in cancer-associated fibroblasts (CAFs) rather than tumor cells. DNA methylation of CERCAM was significantly elevated in metastatic CRC compared to primary tumors. In vitro experiments confirmed that CERCAM was overexpressed in CAFs and CRC tissues. Co-culturing CRC cells (RKO, HCT116) with CERCAM-knockdown CAFs significantly suppressed tumor cell proliferation, migration, and invasion. Furthermore, the ac4C score effectively predicted tumor mutational landscape differences and sensitivities to agents like pazopanib, gefitinib, and conventional chemotherapies.

This study delineates the crucial role of ac4C modification in shaping the CRC microenvironment, identifies CERCAM as a CAF-specific driver of tumor progression, and provides a translational framework for ac4C-based prognostic prediction and personalized targeted therapy.

mRNA-based therapeutics have demonstrated notable success in SARS-CoV-2 vaccines and are emerging in cancer immunotherapy. However, conventional mRNA cancer vaccines are limited in part by the low immunogenicity of tumor-associated and neoantigens. We addressed this limitation by formulating a modular, liposome-based mRNA cocktail comprising three distinct mRNAs encoding tumor antigens, the co-stimulatory molecule CD80, and membrane-tethered IL-2. Administration of this mRNA cocktail resulted in synergistic activation of tumor antigen-specific CD8⁺ T cells and robust anti-tumor immune responses. In addition, substituting IL-2 in the mRNA cocktail with membrane-tethered IL-12 led to the expansion and differentiation of endogenous antigen-specific Th1 helper T cells in vivo. Importantly, this platform activated NY-ESO-1-specific CD8⁺ T cells in HLA-A*02:01-transgenic mice, highlighting its translational potential. This modular mRNA cocktail provides a flexible and translatable platform for precision cancer immunotherapy by enabling coordinated activation of both CD8⁺ and CD4⁺ T cell responses.

Breast cancer (BC) is a disease with a subtle onset, often diagnosed late, and has a high mortality rate. This study aimed to identify long non-coding RNAs (lncRNAs) linked to BC as prognostic markers using bioinformatics and initial experiments, explore their regulatory pathways, and assess their expression and prognostic significance.

Transcriptome data and clinical details of a BC cohort were obtained from The Cancer Genome Atlas and Gene Expression Omnibus. Bioinformatics methods, including Weighted Gene Co-expression Network Analysis (WGCNA), Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), Gene Set Enrichment Analysis (GSEA), and survival analysis, were used to evaluate lncRNAs, related microRNAs (miRNAs), and target genes. The competing endogenous RNA (ceRNA)-based regulatory axis of lncRNA-miRNA-messenger RNA (mRNA) was identified, and gene expression on this axis in normal human mammary epithelial and BC cell lines was analyzed using quantitative reverse transcription polymerase chain reaction (qRT-PCR).

We screened the lncRNA modules most associated with BC traits through bioinformatics analysis and analyzed the immune cell infiltration level of BC samples. Active cluster of differentiation 4+ (CD4+) memory T cells were more infiltrated in BC, whereas resting mast cells were less infiltrated in controls. WGCNA revealed 14 lncRNA modules linked to immune cells and 277 modules associated with BC and immune traits. Univariate Cox analysis identified 12 survival-related target mRNAs mainly involved in immune processes and cell proliferation. We constructed a putative BC-related ceRNA network, specifically the long intergenic non-protein coding RNA 200 (LINC00200)-microRNA 20b (miR-20b-5p)-high mobility group box 3 (HMGB3) axis, through the identification of survival-associated lncRNAs and their target mRNAs to identify key regulatory pathways in BC. qRT-PCR revealed elevated LINC00200 and HMGB3 levels in BC cells, with minor differences in LINC00200 across cell lines. The expression of miR-20b-5p decreased in MCF-7 cells but increased in BT549 and MDA-MB-231.

The study is the first to identify LINC00200 as a potential prognostic lncRNA predictive of survival outcomes in BC and to systematically delineate a novel ceRNA regulatory axis comprising LINC00200, miR-20b-5p, and HMGB3. Although this axis holds promise as a novel prognostic marker, further functional studies and assessment of diagnostic utility are warranted.

Alectinib is a first-line tyrosine kinase inhibitor (TKI) for anaplastic lymphoma kinase (ALK)-rearranged non-small cell lung cancer (NSCLC) and is generally well tolerated. Severe renal toxicity is rare and evidence guiding management in elderly patients is limited. We describe a severe, reversible, dialysis-requiring nephrotoxicity confirmed by drug rechallenge, with successful subsequent treatment using an alternative ALK inhibitor.An elderly patient with advanced ALK-rearranged NSCLC developed kidney injury (AKI) shortly after initiation of alectinib, which progressed to dialysis-requiring renal failure. Renal function improved after treatment discontinuation but deteriorated again on alectinib rechallenge, strongly supporting drug-related causality. Urine sediment findings were consistent with acute tubular necrosis. Alectinib was permanently discontinued and treatment was switched to brigatinib, resulting in stable renal function and sustained oncological response.This case highlights that severe alectinib-induced nephrotoxicity, although rare, can occur and may be life-threatening, particularly in elderly patients with comorbidities and polypharmacy. Importantly, effective disease control can still be achieved through alternative ALK inhibitors.

Small cell lung cancer (SCLC) represents the most lethal subtype of lung carcinoma. Platinum-based agents, such as cisplatin and carboplatin, remain the cornerstone of first-line therapy for patients with SCLC, whether administered as chemotherapy alone or in combination with immunotherapy. However, patients frequently develop platinum resistance within a short time frame, leading to therapeutic failure. The molecular mechanisms underlying platinum resistance in SCLC require further investigation, given that effective intervention strategies remain elusive. Through multiomic analysis integrating transcriptomic data from cisplatin-resistant PDX tissues and the IMpower133 SCLC cohort study and translatomic data from SCLC cells, we identified a platinum resistance-associated noncanonical open reading frame derived from MIR7-3HG. This noncanonical ORF encodes an unannotated 128-amino acid protein, designated SCRMP (SCLC cisplatin resistance-associated microprotein). SCRMP was up-regulated in cisplatin-resistant SCLC tissues and cell lines, and its elevated expression strongly correlated with impaired platinum response and unfavorable survival outcomes in patients. CRISPR-Cas9-mediated SCRMP knockout restored cisplatin sensitivity and promoted apoptosis in SCLC cells, platinum-resistant patient-derived organoids in vitro, and patient-derived xenografts in vivo. Mechanistically, SCRMP potentiated platinum resistance by mediating SUMOylation and nuclear translocation of CSNK2B, which stabilized the RBBP4-p300 complex. This stabilization activated transcription of RAD51C and associated DNA damage repair genes through histone acetylation, ultimately promoting therapeutic resistance. Our study systematically elucidates the biological function of SCRMP in SCLC platinum resistance and delineates its specific molecular mechanism in regulating DNA damage repair, thereby providing theoretical foundations and potential intervention strategies for overcoming platinum resistance in SCLC.