Translational Oncology最新文献

Located on chromosome 5, the THOC3 gene encodes a protein of 351 amino acids, though its precise biological role in various malignancies, including lung cancer (LC), remains to be fully clarified. Elevated expression of THOC3 was observed in LC (IHC) analyses. Increased THOC3 levels were associated with advanced tumor stages and poorer prognosis in patients. Functional studies using both loss-of-function and gain-of-function approaches suggested that THOC3 may influence cellular proliferation, tumorigenic potential, and apoptosis in vitro. Mechanistically, its effects appear to involve activation of the STAT3 pathway. Additionally, Garcinone d-induced STAT3 activation mitigated certain malignant characteristics of LC cells, though silencing THOC3 reversed this effect. These results indicate that THOC3 may serve as a potential prognostic marker and could warrant further investigation as a therapeutic target in LC.

The immunosuppressive tumor microenvironment (TME) and intrinsic heterogeneity of ovarian cancer (OC) are primary drivers of therapeutic resistance and mortality. To deconvolute these complex dynamics and identify robust therapeutic targets, this study employed an integrative strategy combining ensemble machine learning algorithms with high-resolution single-cell transcriptomics and experimental validation. Through dual-feature selection (LASSO and SVM-RFE) applied to multi-cohort bulk transcriptomic data, we identified Stabilin-1 (STAB1) as a top-ranked prognostic determinant. Crucially, single-cell analysis of the OC ecosystem redefined the cellular localization of STAB1, revealing its predominant enrichment in LYVE1+ perivascular-like M2 macrophages and a hyper-aggressive, EMT-active tumor subpopulation. Validating these in silico insights, in vitro loss-of-function assays confirmed that STAB1 silencing in OC cell lines (A2780 and SK-OV-3) significantly suppressed cell proliferation, colony formation, and invasion. Collectively, our findings support STAB1 as a pivotal "dual-checkpoint" molecule that bridges the immunosuppressive stroma and the malignant epithelium, highlighting its potential as a novel therapeutic target for dismantling the ovarian cancer ecosystem.

DNA double-strand breaks (DSBs) are among the most severe forms of DNA damage, and their cellular consequences depend on the efficiency of DNA repair pathways. Non-homologous end joining (NHEJ) provides a rapid mechanism to reconnect broken DNA strands and is a major pathway for maintaining genomic stability. FBXW7α is the substrate-selective unit within the SCF (SKP1-CUL1-F-box) E3 ubiquitin ligase complex, imparting specificity by recruiting proteins destined for ubiquitin-mediated turnover. It has recently been reported to mediate XRCC4 ubiquitylation during DNA damage, and is a recognized tumor suppressor, thereby promoting NHEJ. A circular RNA derived from FBXW7 gene exons 3 and 4 (circ-FBXW7) has been identified. This circRNA encodes a 185-amino acid protein, named FBXW7-185aa, which contains a spanning-junction open reading frame (ORF). We found that compared with FBXW7α, FBXW7-185aa was recruited more slowly to DNA damage sites in glioblastoma (GBM) cells. Functionally, FBXW7-185aa may enhance NHEJ repair by facilitating the removal of FBXW7α from DNA damage lesions, thereby promoting the subsequent steps of DNA repair. Efficient DNA repair can enable tumor cells to survive genotoxic stressors like radiotherapy. Radiotherapy is also known to modulate the tumor immune microenvironment. Therefore, our findings suggest that FBXW7-185aa may represent a potential molecular nexus between DNA damage response and tumor-immune interactions. Moreover, depletion of FBXW7-185aa was associated with increased sensitivity of GBM cells to pevonedistat, a drug that inhibits the ubiquitin-like modification of the protein NEDD8, when combined with radiation. Together, our results indicate that FBXW7-185aa acts as a fine-tuning regulator of NHEJ through its interaction with FBXW7α, adding a new layer to the complexity of FBXW7 gene-derived proteins in DNA repair. These findings provide a mechanistic basis for exploring how fine-tuned DNA repair dynamics may influence GBM radiosensitivity and potentially impact tumor immunogenicity, although direct immunological effects remain to be determined.

Background: Glioblastoma (GBM) is an aggressive primary brain tumor with poor prognosis. Although brain imaging features are related to biological behaviors of GBM, the causal relationship between them remain unclear.

Objectives: To explore the causal relationship between brain imaging features and GBM, identify key pathogenic genes, and provide a perspective for GBM therapy.

Methods: Two-sample MR analysis was employed. Causal relationships were evaluated based on brain imaging features, eQTL, and GWAS data. Differentially expressed brain imaging-related genes were screened through gene mapping and differential expression analysis. MR analysis on eQTL data identified key genes, and GSEA was performed. Given its robust genetic association, high expression in GBM, and enrichment association with tumor malignancy, NRP2 was determined as the core gene, with its function verified by in vitro/in vivo experiments.

Results: MR analysis identified 255 GBM-associated brain imaging features, with 9 key genes selected. NRP2 was identified as a risk gene. NRP2 knockdown significantly inhibited GBM proliferation, migration, and invasion and promoted apoptosis. The inhibitory effects were reversed by activated FAK-signaling pathway. Mechanistically, NRP2 regulated FAK phosphorylation through direct binding, thereby activating the Focal-adhesion pathway and promoting tumor malignancy. In animal experiments, inhibiting NRP2 slowed tumor growth, which was weakened by FAK agonists.

Conclusion: This study establishes the causal relationship between brain imaging features and GBM from a genetic perspective. NRP2 activates Focal-adhesion pathway through FAK signaling to drive GBM progression. NRP2 is a key molecule connecting imaging phenotypes and GBM malignant behaviors, serving as a potential therapeutic target.

Background: Chronic inflammation is a key driver of glioma progression, but its cellular organization within the tumor microenvironment remains poorly understood.

Methods: This study employed an integrated multi-omics analysis strategy, combining bulk transcriptomics, proteomics, and glioma cell line expression data with single-cell RNA sequencing data from paired gliomas and adjacent normal brain tissues. Inflammatory signaling activity was quantitatively assessed using pathway-level scoring methods, endothelial cell heterogeneity was analyzed at single-cell resolution, and pseudo-temporal trajectory analysis and cell-cell communication analysis were further conducted. Immunohistochemical analysis was performed using human glioma brain tissue samples from our center to independently validate key findings at the protein level.

Results: Bulk transcriptomics analysis revealed significantly activated inflammatory signals in glioblastomas. Single-cell analysis identified a highly inflammatory endothelial cell subtype that was significantly enriched in tumor tissues. Cell communication analysis further revealed enhanced signal output capabilities and participation in neurally-related ligand-receptor interactions. CEP135 was specifically enriched in this endothelial cell subtype and showed consistent upregulation of both transcriptional and protein levels across multiple independent datasets. Immunohistochemical analysis of glioma brain tissue from our center confirmed that CEP135 is primarily localized in tumor-associated endothelial regions, and its expression level was significantly correlated with increasing tumor grade.High CEP135 expression was associated with poor treatment response, shorter survival outcomes.

Conclusion: This study identified a innovative CEP135+inflammation-associated endothelial cell subtype and established CEP135 as a key biomarker linking endothelial inflammation reprogramming, tumor progression, and adverse clinical outcomes.

Objective: Transcriptomic subtyping is not yet standardized for prognostic use in epithelial ovarian cancer (EOC). This study aims to validate RNA sequencing (RNAseq) from formalin-fixed, paraffin-embedded (FFPE) tissues and to evaluate survival-associated transcriptomic subtypes and differentially expressed genes (DEGs) in a clinical trial cohort.

Methods: An exploratory post hoc analysis was conducted on FFPE samples from patients enrolled in the ENGOT-ov24/NSGO-AVANOVA1&2 trial. RNA was extracted and sequenced, and gene expression analysis was performed to classify subtypes using established, microarray-based, algorithms. Differentially expressed genes (DEGs) were identified based on survival groups, and survival outcomes were analyzed using Kaplan-Meier curves.

Results: Of 96 eligible samples, 82 were included in the final analysis. Subtype classifications showed moderate agreement across RNAseq data formats. However, gene expression variability showed inconsistent concordance with clinical metadata and molecular subtypes. Eighteen genes were differentially expressed between long- and short-term survivors. Notably, DPEP3 and SLC14A1, were significantly upregulated in long-term survivors. Despite distinct expression patterns, no significant survival differences were observed between subtypes.

Conclusions: This study demonstrates the feasibility of using RNAseq on FFPE tissue in EOC, while also highlighting challenges of applying microarray-based transcriptomic subtypes to RNAseq data. Transcriptomic analysis identified potential prognostic gene candidates but also highlighted the need to refine classification tools. Further research is essential to improve the molecular classification of EOC, thereby enhancing prognostic accuracy and guiding future therapeutic strategies.

Background: l-type calcium channels (LTCCs) play an important role in tumorigenesis, but their expression profile, functional significance, and therapeutic potential in gastric adenocarcinoma (GAC) remain unclear. Concurrently, aberrant lipid metabolism is widely acknowledged as a key driver of GAC progression, but its upstream regulatory mechanisms have yet to be fully elucidated. This study was designed to explore the association between LTCCs and lipid metabolic reprogramming in GAC and to investigate its influence on GAC progression and clinical outcomes.

Methods: The expression profiles of LTCCs family members in GAC were analyzed using the TCGA database. Gastric cancer cells were treated with the LTCCs antagonist (+)-Bay-K-8644, and its effect on the malignant phenotype of the cells was detected by CCK-8, clone formation, scratch healing and transwell assay. Metabolic changes were analyzed by UPLC-Q-TOF-MS metabolomics. The role of lipid metabolism in the regulation of LTCCs was verified by palmitic acid (PA) backfill assay.

Results: Bioinformatics analysis showed that Voltage-dependent LTCCs subunit alpha-1D (CACNA1D) was up-regulated in GAC. CACNA1D had excellent diagnostic value and its high expression was associated with poor prognosis. (+)-Bay-K-8644 significantly inhibited gastric cancer cells' proliferation, clone formation, and migration ability. Metabolomics analysis revealed that (+)-Bay-K-8644 treatment resulted in reprogramming of lipid metabolism. Exogenous PA was able to partially reverse the anticancer effects of (+)-Bay-K-8644 and restore cell proliferation and migration.

Conclusions: This study elucidated the unique expression pattern of LTCCs in GAC for the first time, and confirmed that LTCCs antagonism exerts anticancer effects by inducing reprogramming of lipid metabolism. These findings provide a theoretical basis for repositioning calcium channel blockers for GAC therapy and suggest that combined targeting of calcium channels and lipid metabolism may become a new strategy for GAC treatment.

Objective: The tumor microenvironment (TME) of hepatocellular carcinoma (HCC) is shaped by underlying liver pathology, with potentially distinct features in HCC without cirrhosis (Non-cirrHCC) compared to HCC with cirrhosis (CirrHCC); however, these background-specific differences remain incompletely understood. This study aimed to systematically characterize and compare the cellular composition, spatial organization, and immunoregulatory interactions of the TME between Non-cirrHCC and CirrHCC.

Methods: Total 278 HBV-positive cases and mapped 2,837,999 cells across 11 major cell types were applied for spatial analysis. Subsequently, we integrated the spatial data with publicly available single-cell RNA-seq datasets for downstream analysis.

Results: Comparative analysis demonstrated marked differences in immune cell composition between Non-cirrHCC and CirrHCC, with CirrHCC characterized by a pronounced decline in functionally active CD8⁺ T cells. We identified 10 distinct heterotypic cellular neighborhoods (HCNs) representing the heterotypic architecture of the tumor microenvironment. Notably, CirrHCC exhibited an immunosuppressive microenvironment with increased spatial proximity between Tregs and CD8T_CD107a⁺ cells, leading to reduced CD8⁺ T cell functional signaling. Integration with single-cell RNA sequencing from public database further indicated that, in CirrHCC, Tregs preferentially interact with the CD8T_CD107a+ cells, potentially mediated by the SPP1-ITGA4 signaling axis.

Conclusion: In conclusion, CirrHCC promotes a spatially organized Treg-CD8T_CD107a⁺ suppressive niche that constrains CD8⁺ T cell effector function in HCC, with SPP1-ITGA4 emerging as a potential mediating pathway.

Background: Retroperitoneal sarcomas (RPS) exhibit a high locoregional recurrence rate after macroscopically complete surgical resection. Systemic therapies have limited efficacy and significant adverse effects. Sarcoma cell lines are susceptible to paclitaxel (PTX), a microtubule stabilizer, in 2-dimensional (2-D) monolayer cell culture, but resistant in animal models. When locally delivered via drug-eluting buttresses supratherapeutic concentrations are achieved and PTX becomes efficacious. Due to the limitations of 2-D culture, we establish an organotypic culture system to model the mechanisms by which supratherapeutic and prolonged exposure of PTX is effective.

Methods: Liposarcoma tumors (LP6) were established subcutaneously in NU/J mice. Tumors were harvested, sliced with a vibratome (250 µm thick), and cultured on permeable trans wells.

Results: Organotypic culture viability was maintained up to 7 days with greater than 50 % viability. Tumor slices were composed of 82 % ± 7 % human liposarcoma cells with the remainder being mouse stroma as determined by CD44 staining. Under 4-day exposure, IC₅₀ of PTX with organotypic culture shifted 7000 rightward as compared to 2-D culture. A subset of 17 genes was significantly differentially expressed as compared to untreated controls, while no genes were differentially expressed after 1 day of treatment. Gene set enrichment analysis demonstrated enrichment in apoptotic, extracellular matrix, cell motility, and cell cycle pathways. Caspase-8 activation occurred only at 10,000 ng/mL and 4-days of PTX or greater.

Conclusion: This study reports a reproducible, clinically relevant organotypic culture liposarcoma model, which can serve as an intermediary between in vitro and in vivo studies.

Background: Resistance to temozolomide (TMZ)-based chemotherapy is a major cause of progression and recurrence in patients with glioblastoma. Although the deubiquitinating enzyme ubiquitin C-terminal hydrolase L1 (UCHL1) has been implicated in tumor chemoresistance, its roles and underlying mechanisms in TMZ resistance remain unclear.

Methods: Bioinformatics analyses and immunohistochemistry were used to assess the prognostic significance and the levels of UCHL1 in glioma specimens. Co-immunoprecipitation assays and mass spectrometry were performed to identify protein interactors of UCHL1. The impact of the UCHL1-Keratin 8 (KRT8) axis on TMZ resistance was evaluated using the cell counting kit-8 (CCK-8) assay and western blotting.

Results: We found that UCHL1 induced resistance of glioblastoma cells to TMZ. Moreover, UCHL1 expression was significantly upregulated in patients with TMZ-resistant glioma and correlated with poor prognosis. As a deubiquitinase belonging to the UCH enzyme family, UCHL1 utilizes its deubiquitination activity to remove K27-linked polyubiquitin chains from KRT8, thereby preventing KRT8 degradation by the proteasome and maintaining its stability. In vitro assays further demonstrated that depletion of the UCHL1-KRT8 axis weakened chemoresistance by increasing TMZ-induced apoptosis.

Conclusion: These findings reveal a novel signaling axis underlying TMZ resistance in glioblastoma and highlight the UCHL1-KRT8 axis as a promising therapeutic target.