Multi-omics integration and functional validation identify thrombospondin-1 as a functionally relevant mediator of doramectin-associated inhibition of glioblastoma migration and invasion.

NTMT1 (METTL11A) promotes MYC-dependent proliferation and downstream HLA-A suppression in cervical cancer

Background Identifying upstream molecular regulators linking tumor proliferation with immune-related alterations remains an important challenge in cancer therapy. NTMT1 (METTL11A), a protein N-terminal methyltransferase, has been implicated in tumorigenesis; however, its role in coordinating tumor–immune interaction is poorly understood. Methods We performed integrative single-cell RNA sequencing (GSE208653) and spatial transcriptomics (GSE208654) analyses to characterize NTMT1 expression and function in cervical cancer. Spatial deconvolution and microenvironmental co-localization analyses were used to define context-specific effects. Functional validation was conducted using RT–qPCR, Western blotting, multiplex immunofluorescence, and flow cytometry-based assays. Results NTMT1 exhibited heterogeneous expression across epithelial and immune cell populations, with enrichment in squamous cell carcinoma. Spatial transcriptomics revealed that NTMT1-positive regions were associated with altered immune composition, including reduced macrophage and increased NK/T cell infiltration. In epithelial-enriched regions, NTMT1 expression correlated with activation of cell cycle pathways, including MYC, E2F1, CDK1, and CCNB1. Functional experiments demonstrated that NTMT1 promotes cell cycle progression via MYC upregulation. In epithelial–NK/T co-localization niches, NTMT1 was associated with modulation of antigen presentation pathways and suppression of HLA-A expression. Functionally, NTMT1 overexpression reduced IFN-γ production by CD8+ T cells under the co-culture conditions used in this study, which was partially restored by HLA-A re-expression. Conclusion NTMT1 was identified as a candidate regulator associated with MYC activation and HLA-A suppression. Although the precise molecular mechanism remains to be elucidated, functional experiments demonstrated that NTMT1 overexpression was accompanied by increased MYC expression, reduced HLA-A expression, enhanced cell-cycle progression, and impaired CD8+ T-cell function. These findings identify NTMT1 as a candidate regulatory node associated with MYC activation and downstream HLA-A suppression, warranting further mechanistic and in-vivo investigation, highlighting NTMT1 as a promising candidate for future therapeutic investigation. Further in vivo studies will be required to determine its suitability as a target for combination immunotherapy.

Unraveling the role of PPFIA1 in cancer: a comprehensive multi-omics analysis and functional validation from pan-cancer to pancreatic cancer

Background PTPRF interacting protein alpha 1 (PPFIA1), a cytoplasmic scaffold protein of the liprin family, modulates cell adhesion, signal transduction, and cytoskeletal dynamics. Its pan-cancer prognostic implications and functional roles, particularly in pancreatic cancer, remain underexplored. Methods Pan-cancer datasets including TCGA, GTEx and TISCH were integrated to assess PPFIA1 expression, genomic alterations, and pathway enrichment via Kaplan-Meier survival analysis, Cox regression, cBioPortal, and GSEA. In pancreatic cancer, external cohorts (e.g., GSE28735, GSE52452) were used for expression and prognosis validation. Besides, immunotherapy response was analyzed by using TIGER cohorts. Functional impacts were determined by using ShRNA-mediated knockdown in CFPAC-1, PANC-1 and Panc-02 cells and evaluating their proliferation, migration and invasion in vitro and in vivo. Results PPFIA1 exhibited differential expression across multiple cancer types, with overexpression in pancreatic adenocarcinoma (PAAD) versus non-tumor tissue at mRNA/protein levels, predominantly in malignant cells. High PPFIA1 correlated with adverse prognosis in PAAD across multiple endpoints. Genomic analyses revealed amplifications in head/neck cancers and mutations in endometrial carcinoma, clustering in SAM domains. GSEA enrichment analysis indicated pan-cancer activation of the mitotic spindle assembly pathway and Epithelial-Mesenchymal Transition (EMT) pathways in PAAD. Knockdown suppressed PAAD cell proliferation, colony formation, migration, invasion, and tumor growth in vivo. Notably, elevated PPFIA1 predicted superior responses to immunotherapy in pan-cancer cohorts rather than pancreatic cancer. Conclusions PPFIA1 emerges as a pan-cancer biomarker with prognostic significance and shows an oncogenic role in PAAD, potentially associating with EMT-related malignant phenotypes. Its association with immunotherapy efficacy suggests PPFIA1 warrants further investigation as a candidate biomarker and potential functional target for precision oncology.

Identification and validation of NCOA5-ZCCHC3/FGF22 axis as a prognostic marker in lung adenocarcinoma migration and invasion

Lung adenocarcinoma (LUAD), the most common subtype of lung cancer, is associated with substantial global mortality. Nuclear receptor coactivator 5 (NCOA5) has been implicated in several malignancies; however, its functional role and regulatory mechanisms in LUAD remain largely unknown. In this study, NCOA5 expression was evaluated in 94 paired LUAD and adjacent tissues using immunohistochemistry, RT-PCR, and Western blotting. Functional analyses were conducted using shRNA knockdown, CRISPR-mediated knockout, and overexpression models to assess the effects of NCOA5 on LUAD cell proliferation, migration, invasion, apoptosis, cell cycle progression, and organoid formation. Xenograft models were used to validate tumorigenicity in vivo. IP-MS and co-immunoprecipitation identified NCOA5-interacting proteins, while ChIP-seq and dual-luciferase assays interrogated downstream transcriptional regulation. NCOA5 was significantly upregulated in LUAD tissues and associated with advanced stage, poor differentiation, and reduced overall survival, serving as an independent prognostic factor ( P  < .040). NCOA5 knockdown inhibited LUAD cell proliferation, migration, invasion, induced G0/G1 arrest, promoted apoptosis, reduced organoid formation, and suppressed xenograft growth, whereas NCOA5 overexpression produced the opposite effects. Among 39 candidate interacting proteins identified by IP-MS, ZCCHC3 was validated as a direct NCOA5-binding partner. ZCCHC3 depletion phenocopied NCOA5 loss and reversed NCOA5-induced proliferation, migration, invasion, and colony formation, supporting a functional NCOA5–ZCCHC3 interaction. ChIP-seq analysis identified fibroblast growth factor 22 (FGF22) as a direct transcriptional target of NCOA5. FGF22 was markedly downregulated in LUAD tissues and higher FGF22 expression was associated with improved patient survival. NCOA5 suppressed FGF22 transcription, while NCOA5 inhibition increased FGF22 expression. Functionally, FGF22 knockdown enhanced LUAD aggressiveness, whereas FGF22 restoration abrogated the oncogenic effects of NCOA5 in vitro and in vivo. Collectively, these findings identify a previously unrecognized NCOA5–ZCCHC3/FGF22 axis that drives LUAD progression and provide new mechanistic insight into the role of NCOA5 in LUAD biology.

FZD5 drives macrophage-mediated immunomodulation and predicts prognosis in glioma: evidence from single-cell sequencing.

BACKGROUND Gliomas are highly malignant brain tumors characterized by an immunosuppressive microenvironment, which limits therapeutic efficacy and contributes to poor clinical outcomes. The WNT/β-catenin signaling pathway is critically involved in tumor progression, and FZD5, a key receptor within this pathway, may participate in immune regulation. However, its specific role and underlying mechanisms in glioma remain unclear. METHODS RNA-seq and microarray datasets from the Chinese Glioma Genome Atlas (CGGA) and The Cancer Genome Atlas (TCGA), together with single-cell RNA sequencing (scRNA-seq) datasets from GEO, were comprehensively analyzed. The Seurat package was used to identify macrophage-related clusters and mitophagy-associated pathways. Cox and LASSO regression analyses, along with a prognostic nomogram, were applied to evaluate the prognostic significance of FZD5. Immune infiltration, functional enrichment, and immunotherapy response analyses were conducted, followed by validation using spatial transcriptomics, immunohistochemistry, and in vitro assays. RESULTS In bulk glioma transcriptomes, FZD5 emerged as an independent predictor of poor prognosis. Crucially, single-cell and spatial analyses revealed that the biologically significant FZD5 signal originated predominantly within tumor-associated macrophages (TAMs), where it colocalized with the M2 marker CD163. Consistently, elevated FZD5 levels correlated with increased myeloid infiltration and an immunosuppressive tumor microenvironment. Functionally, macrophage-expressed FZD5 was associated with mitophagy-related programs and promoted an M2-skewed phenotype, thereby enhancing glioma cell proliferation, migration, and invasion via macrophage-glioma crosstalk. CONCLUSION FZD5 is a TAM-enriched marker in glioma tissues and a potential regulator of macrophage-associated immunosuppressive programs, supporting its utility as a prognostic biomarker and a candidate target for microenvironment-oriented interventions in glioma.

Integrative multi-omics profiling reveals DRG1 promotes hepatocellular carcinoma via c-Myc phosphorylation and facilitates lenvatinib resistance.

BACKGROUND & AIMS Developmentally regulated GTP-binding protein 1 (DRG1), a member of the GTPase family, has been implicated in cancer progression; however, its biological function and therapeutic relevance in hepatocellular carcinoma (HCC) remain poorly understood. This study aimed to define the role of DRG1 in HCC progression and uncover its underlying regulatory mechanisms. METHODS Multi-omics analyses, clinical tissue validation, in vitro functional assays, and in vivo xenograft models were integrated to investigate the biological function, regulatory mechanism, and therapeutic relevance of DRG1 in HCC. RESULTS DRG1 was significantly upregulated in HCC tissues and associated with aggressive clinicopathological characteristics and poor patient survival. Integrated single-cell and spatial transcriptomic analyses revealed preferential DRG1 expression in malignant regions and its association with an immunosuppressive tumor microenvironment. Mechanistically, DRG1 promoted HCC cell proliferation through regulation of c-Myc activation, while HDAC2 was identified as an upstream regulator of DRG1 expression. Furthermore, DRG1 depletion enhanced sensitivity to lenvatinib treatment in HCC models. CONCLUSIONS Our findings identify DRG1 as a potential prognostic biomarker and therapeutic target in HCC. The HDAC2-DRG1-c-Myc regulatory network contributes to malignant progression, while DRG1 inhibition represents a potential strategy to improve lenvatinib response.

Transcriptomic Profiling of Adhesion GPCRs in Pancreatic Adenocarcinoma Identifies ADGRG7 and ADGRF1 as Novel Diagnostic Biomarkers and Therapeutic Targets