TL;DR
High MCT4 expression in TAMs may be associated with metabolic reprogramming toward glycolysis, and could promote M2 polarization of TAMs, thereby contributing to HCC progression and poor clinical outcomes.
In hepatocellular carcinoma (HCC), the polarization of tumor-associated macrophages (TAMs) towards the M2 phenotype is a key factor driving the formation of an immunosuppressive microenvironment and malignant progression, a process frequently associated with glycolytic metabolic reprogramming. Nucleolar and spindle associated protein 1 (NUSAP1) is aberrantly overexpressed in various malignancies. However, whether it promotes HCC progression by driving TAM M2 polarization through regulating tumor cell glycolysis remains unclear. Bioinformatics analysis was used to examine NUSAP1 expression in HCC and its correlation with M2 TAM infiltration. qRT-PCR and Western blot were employed to validate NUSAP1 expression levels in HCC cell lines. Functional experiments, including flow cytometry, ELISA, colony formation, and Transwell assays, were conducted to investigate the regulatory effects of NUSAP1 on TAM M2 polarization and HCC cell malignant behaviors. GSEA and correlation analyses were performed to explore the association between NUSAP1 and the glycolysis pathway. The impact of NUSAP1 on glycolytic metabolic reprogramming in HCC cells was assessed via Seahorse energy metabolism analysis, ATP/lactate production assays, and detection of key glycolytic protein expression. Furthermore, bioinformatics analysis predicted the transcriptional regulation of NUSAP1 by forkhead box P1 (FOXP1), which was subsequently validated by ChIP and dual-luciferase reporter assays. Finally, in vitro rescue experiments and an allograft mouse tumor model were used to confirm that the FOXP1/NUSAP1 axis induces TAM M2 polarization via glycolysis and accelerates HCC progression. NUSAP1 was significantly overexpressed in HCC, and its expression level positively correlated with M2 TAM infiltration. Functional experiments demonstrated that NUSAP1 facilitates HCC progression by driving glycolytic metabolic reprogramming in tumor cells and inducing polarization to the M2 phenotype. Mechanistically, FOXP1, as a transcription factor, binds to the NUSAP1 promoter region and activates its transcription. Both in vitro and in vivo experiments further confirmed that the FOXP1/NUSAP1 axis promotes TAM M2 polarization by activating glycolytic metabolism, thereby accelerating HCC progression. This study reveals a novel mechanism by which FOXP1 transcriptionally upregulates NUSAP1 expression, resulting in glycolytic metabolic reprogramming that induces TAM M2 polarization and promotes HCC progression. These findings identify a potential therapeutic avenue for reversing the immunosuppressive microenvironment in HCC.
Cancer Stem Cells (CSCs) are the core drivers of esophageal cancer recurrence, metastasis, and treatment resistance, and their metabolic reprogramming characteristics provide an important entry point for targeted therapy. This study aimed to explore the role and regulatory mechanism of HACD1 (3-Hydroxyacyl-CoA Dehydrogenase 1), a key enzyme in lipid metabolism, in maintaining the stemness of esophageal cancer. Using the Patient-Derived Xenograft (PDX) cell line HD-12 and laboratory cell lines TE-1 and KYSE-150, a Tumoursphere (TS) stemness enrichment model was constructed, and systematic analysis was conducted by combining quantitative Polymerase Chain Reaction (qPCR), transcriptome sequencing, lipidomics, functional experiments, and animal models. The results showed that the upregulation of stemness markers (Oct4(Octamer-binding transcription factor 4), Sox2 (SRY-box transcription factor 2), etc.) in PDX-derived HD-12 tumourspheres was significantly higher than that in laboratory cell lines, with unique transcriptomic characteristics. Lipidomics confirmed that lipid metabolites such as Triglycerides (TG) and Long-Chain Fatty Acids (LCFA) were significantly enriched in tumourspheres. The HACD (3-Hydroxyacyl-CoA Dehydrogenase) family was specifically highly expressed in stem cells, and the enzymatic activity of HACD1 (3-hydroxyacyl-CoA dehydratase 1) was positively correlated with the stemness phenotype. HACD1 knockdown significantly inhibited esophageal cancer tumoursphere formation, cell migration, and in vivo tumourigenic ability, and downregulated the expression of stemness markers; exogenous supplementation of Docosahexaenoic Acid (DA) could reverse this inhibitory effect in a concentration-dependent manner. Metabolic analysis indicated that tumoursphere cells rely on both glycolysis and lipid oxidation for energy supply and possess metabolic flexibility, and blocking a single pathway cannot effectively inhibit stemness. This study is the first to confirm that HACD1 maintains the stemness phenotype of esophageal cancer by regulating lipid metabolism. The PDX model exhibits a unique stemness regulatory pattern closer to clinical practice, providing new molecular targets and experimental basis for the metabolic targeted therapy of esophageal cancer.
BACKGROUND Hepatocellular carcinoma (HCC) arises within a hypoxic and immunosuppressive tumor microenvironment (TME), where tumor-associated macrophages (TAMs) constitute a major immune population. The impact of hypoxia on TAM functional heterogeneity and their contribution to HCC growth and metastasis remain incompletely understood. METHODS We integrated multiple single-cell RNA sequencing datasets and developed a machine learning framework to map cellular hypoxia at single-cell resolution. Hypoxic TAMs (H-TAM) were characterized using transcriptomic and functional assays, including pseudotime trajectory analysis, regulatory network inference, in vitro co-culture, and orthotopic mouse models. Drug sensitivity correlations and in vivo validation were performed to evaluate therapeutic strategies. RESULTS H-TAM represented the most hypoxic immune population in HCC and exhibited enhanced interactions with malignant hepatocytes. Transcriptomic profiling revealed HIF-1α-dependent hypoxia signaling and upregulation of plasminogen activator, urokinase (uPA, encoded by PLAU). H-TAM-derived uPA engaged its receptor uPAR on HCC cells, promoting epithelial-mesenchymal transition (EMT), migration, invasion, and lung metastasis. In line with this, genetic silencing of the corresponding mouse gene Plau in hypoxia-exposed bone marrow-derived macrophages (H-BMDM) markedly attenuated these pro-tumorigenic effects. Integrative drug sensitivity analysis identified dasatinib as a potential therapeutic agent in HCC with high uPAR expression, and in vivo administration selectively suppressed H-BMDM mediated tumor progression and metastasis while prolonging mouse survival. CONCLUSIONS Hypoxia drives TAM heterogeneity in HCC via an H‑TAM‑intrinsic HIF‑1α-uPA axis that engages uPAR on HCC cells to promote metastasis. uPAR is a potential prognostic biomarker, and dasatinib is a promising therapy to block this axis and improve outcomes.
It is proposed SLC25A1 is associated with metabolic reprogramming in HCC and contribute to tumor progression by promoting lipogenesis and offering a promising therapeutic target for early intervention and treatment.
BACKGROUND Metabolic reprogramming represents a hallmark feature of hepatocellular carcinoma (HCC). As a crucial branch of the polyol pathway, the biological functions and clinical implications of sorbitol metabolism in HCC progression remain to be fully elucidated. METHODS This study integrated single-cell transcriptomic data from 67 HCC patients to establish a sorbitol metabolism scoring system. Pseudotime trajectory analysis was employed to investigate the differentiation patterns of cells with aberrant sorbitol metabolism, while spatial transcriptomics was utilized to characterize their spatial distribution. Using machine learning approaches on HCC cohort transcriptomic data, we developed a prognostic prediction model, complemented by CIBERSORT-based immune infiltration analysis and CellMiner-derived drug sensitivity predictions. In addition, in vitro gain- and loss-of-function experiments were conducted to validate the biological role of SQSTM1 in HCC cells. RESULTS Our findings demonstrate that ALDH3A1+ malignant cells with high sorbitol metabolism scores exhibit dysregulated cell adhesion, enhanced immune evasion capacity, and activated hypoxia signaling pathways. These cells were predominantly localized at the tumor invasive front. The ALDH3A1+-based predictive model identified a high-risk group with significantly poorer prognosis, characterized by increased TP53 mutation frequency and a distinct immunosuppressive microenvironment. Drug sensitivity analysis suggested Irofulven as a potential therapeutic agent targeting sorbitol metabolism-active tumor cells. Furthermore, in vitro experiments demonstrated that SQSTM1 promoted HCC cell proliferation and migration, supporting its functional involvement in HCC progression. CONCLUSION This study uncovers the mechanistic role of sorbitol metabolism in driving HCC progression through shaping an immunosuppressive microenvironment. The sorbitol metabolism scoring system may serve as a novel prognostic biomarker and provides a theoretical foundation for precision treatment strategies, such as Irofulven-targeted therapy, in HCC management. Moreover, SQSTM1 may act as an important downstream functional mediator and represents a potential therapeutic target in hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) poses a significant global health burden with limited therapeutic options, particularly for non-viral etiologies. The mitochondrial solute carrier SLC25A43 is implicated in cellular redox homeostasis, yet its role in HCC remains unclear. This study aimed to comprehensively investigate the expression pattern, clinical significance, biological function, and potential mechanisms of SLC25A43 in HCC. Utilizing multi-omics data from public databases (TCGA-LIHC, GEO, and HPA), we performed integrated bioinformatic analyses. SLC25A43 was consistently upregulated in HCC tissues compared with non-tumorous liver tissues and demonstrated strong diagnostic value (AUC = 0.861). High SLC25A43 expression was significantly associated with advanced tumor stage, metastasis, and adverse clinicopathological features. Survival analyses identified SLC25A43 as an independent prognostic risk factor for overall survival, progression-free interval, and disease-specific survival. Functional enrichment analyses suggested that SLC25A43 is involved in mitochondrial oxidative phosphorylation, energy metabolism, and immune-related pathways. Immune infiltration analyses using ssGSEA, xCell, and TIMER consistently revealed negative correlations between SLC25A43 expression and multiple antitumor immune cell populations, particularly CD8 + T cells. Experimental validation confirmed that SLC25A43 was significantly upregulated in HCC tissues at both mRNA and protein levels. Functional assays in Huh-7, Hep-LM3, MHCC97H, and LO2 cells demonstrated that SLC25A43 knockdown inhibited, whereas overexpression promoted, cell proliferation and migration. Rescue experiments further verified the specificity of these effects. Mechanistically, SLC25A43 regulated intracellular ATP production, ROS accumulation, and glutathione metabolism, indicating a role in redox homeostasis and energy metabolism. In addition, PBMC co-culture experiments showed that SLC25A43 suppressed CD8 + T-cell cytotoxic activity by reducing Granzyme B expression. A prognostic nomogram incorporating SLC25A43 exhibited favorable predictive performance and was successfully validated in two independent GEO cohorts. SLC25A43 is a novel diagnostic and prognostic biomarker for HCC. Its upregulation promotes tumor progression through metabolic reprogramming, redox homeostasis remodeling, and suppression of antitumor immune responses. These findings highlight SLC25A43 as a promising therapeutic target and provide new insights into the metabolic-immune regulatory network in hepatocellular carcinoma.