Abstract Ferroptosis has increasingly emerged as a novel target for cancer therapy because of intrinsic or acquired ferroptosis vulnerabilities in cancers. Small nucleolar RNA (snoRNA)-mediated mRNA modifications play a critical role in regulating ferroptosis and supporting tumor cell adaptation. However, the function and mechanism of the box C/D snoRNA SNORD60 in hepatoblastoma (HB) remain poorly understood. Here, we found that SNORD60 expression was markedly downregulated in HB tissues, and restoring its expression inhibited HB cell proliferation and induced ferroptosis both in vitro and in vivo. Mechanistically, SNORD60 guided the 2′-O-methylation (Nm) of kielin/chordin-like protein (KCP) mRNA, and accelerated its degradation by resolving a G-quadruplex proximal to the Nm site. Furthermore, Silencing KCP induced lipid peroxidation and ferroptosis by suppressing ATF4-mediated transcription of SLC7A11. Moreover, plasma KCP protein levels were markedly higher in patients with HB than in healthy individuals, supporting KCP as a reliable non-invasive diagnostic biomarker for HB. High KCP expression correlates with poor patient prognosis. Overall, our findings reveal that SNORD60-mediated Nm modification of KCP prevents tumor cells from evading ferroptosis and inhibits tumor progression. This SNORD60/KCP axis represents a novel ferroptosis vulnerability and a promising therapeutic target in HB.

Mitochondrial dysfunction has been increasingly recognized as a key pathophysiological contributor to congenital heart disease (CHD), yet its underlying genetic causal effects remain incompletely characterized. This study aimed to investigate the genetically supported associations between mitochondrial-related genes and CHD under a multi-omics framework. We integrated three-layer quantitative trait loci (QTL) data, including DNA methylation, gene expression and protein abundance, of 1,123 nuclear-encoded mitochondrial genes with genome-wide association study (GWAS) summary statistics of overall CHD from FinnGen R12. Summary-based Mendelian randomization (SMR) was applied to systematically evaluate associations between mitochondrial-related molecular traits and CHD. Heterogeneity in dependent instruments (HEIDI) test, colocalization analysis and single-nucleus transcriptomic analysis (GSE203274) were further performed to improve the reliability of identified candidate genes. After integrating three-layer multi-omics data, we identified 13 candidate genes associated with CHD susceptibility with evidence across at least two layers. These genes were predominantly involved in mitochondrial DNA (mtDNA) replication, mitochondrial translation, and mitochondrial respiratory chain, indicating that these pathways may mediate the pathogenic association between mitochondrial dysfunction and CHD. Among these promising genes, POLG and EARS2 were classified as Tier 1 candidates, with consistent directional associations observed in both peripheral blood and cardiac-specific datasets. POLG encodes the sole DNA polymerase responsible for mtDNA replication and repair, while EARS2 functions as a mitochondrial glutamate-tRNA synthetase to mediate the translation of respiratory chain complex subunits. Specifically, hypermethylation at cg04001880 and cg22187094 was associated with higher POLG expression and increased CHD susceptibility, whereas hypermethylation at cg03187795 was linked to lower EARS2 expression and reduced CHD-related risk. Independent single-nucleus transcriptomic data (GSE203274) provided suggestive supporting evidence for partial regulatory patterns observed in our multi-omics analyses. Our multi-omics SMR analysis prioritizes POLG and EARS2 as candidate genes genetically implicated in the susceptibility of overall CHD, and further adds evidence supporting a potential role for mitochondrial dysfunction in CHD etiology. These findings enhance our understanding of the complex genetic mechanisms of CHD and provide promising molecular targets for subsequent mechanistic investigation and therapeutic development.

Transfer RNA (tRNA) modifications play a critical role in regulating codon-specific mRNA translation and enabling tumor cell adaptation. The RNA methyltransferase METTL1 installs N7-methylguanosine (m⁷G) modifications on tRNAs, thereby shaping codon usage and translational output. However, the function and mechanistic contribution of the METTL1-tRNA axis in pancreatic ductal adenocarcinoma (PDAC) remain poorly defined. Here, we show that METTL1 is overexpressed in PDAC tissues and that elevated METTL1 expression is associated with poor patient survival. Genetic ablation of METTL1 markedly suppresses PDAC cell proliferation, migration, and tumor growth in vitro and in vivo. Mechanistically, METTL1 loss selectively reduces m⁷G-modified valine tRNAs - particularly, Val-AAC, Val-CAC, and Val-TAC - leading to impaired translation of valine-enriched oxidative phosphorylation transcripts. As a consequence, METTL1 deficiency disrupts mitochondrial respiration and energy production in PDAC cells. Consistent with this model, valine tRNA levels are elevated in PDAC tissues, and their selective depletion phenocopies METTL1 loss by impairing mitochondrial bioenergetics and tumor cell fitness. Thus, the METTL1-valine tRNA axis promotes PDAC progression through codon-dependent translational control of mitochondrial electron transport chain and oxidative metabolism. Together, our findings identify a METTL1-tRNA-mitochondrial signaling axis as a previously unrecognized metabolic vulnerability and a promising therapeutic target in pancreatic cancer.

Synthesis of the extracellular matrix provides a novel opportunity to cell behavior control. However, exploration of the application of nucleic acid materials for cell surface engineering to achieve physical regulation of migration remains relatively limited. In this study, a self-assembled DNA network anchored to the cell membrane was constructed as a synthetic matrix mimic, which has been shown to inhibit cancer cell migration and invasion through physical confinement. The DNA network forms a dense and uniform engineering on the surface of tumor cells through a process of self-assembly. This study found that coating with RCA1/2 network led to a significant reduction in wound healing rate of approximately 49.19% and invasion rate of approximately 44.49%. The study also indicates that the potential mechanism for inhibiting cell migration and invasion is that coating of the DNA network on the membrane significantly limits membrane fluidity and promotes integrin retention on the membrane, thus interfering with its dynamic circulation. The spatial confinement in this study provides a novel DNA-based platform for controlling cell behavior as well as opening up a new paradigm for anti-migratory and anti-invasive strategies.

Accurate detection of single-nucleotide variants (SNV) is critical for genetic disease diagnosis and cancer profiling, yet existing methods often lack the combination of specificity, multiplexing capacity, and rapid workflow required for clinical application. Here, we report a ligase-mediated encoding (LmCo) system that integrates ligase chain reaction (LCR) for single-base discrimination with hybridization chain reaction (HCR) for signal amplification and multicolor fluorescence encoding. The system achieves a detection limit of 0.1626 fM and enables reliable identification of mutant alleles at frequencies as low as 0.0001% variant allele fraction, substantially outperforming conventional methods. LmCo simultaneously detects six or more SNV targets in a single closed-tube reaction with minimal crosstalk, demonstrates robust stability in complex matrices, and achieves 96.7% accuracy in simulated clinical samples. This platform provides a rapid, sensitive, and highly multiplexed solution for SNV detection in molecular diagnostics and precision medicine.

Abstract Background: Transfer RNA (tRNA) modifications play a critical role in regulating codon-specific mRNA translation and supporting tumor cell adaptation. The RNA methyltransferase METTL1 installs N7-methylguanosine (m7G) modifications on tRNAs, shaping their codon usage landscape and influencing protein synthesis. However, the function and mechanism of the METTL1/tRNA axis in pancreatic ductal adenocarcinoma (PDAC) remain poorly understood. This study examines the role of METTL1-mediated m7G tRNA modification in the progression of PDAC and its associated metabolic reprogramming. Methods: METTL1 expression was analyzed using transcriptomic data from The Cancer Genome Atlas (TCGA) and validated in an independent PDAC tissue cohort by qRT-PCR. METTL1-knockout PDAC cell lines were generated via CRISPR/Cas9. Cellular proliferation and migration were assessed through MTT, colony formation, and wound-healing assays, while tumor growth was evaluated in xenograft models. Global tRNA m7G levels were measured using dot blot, and qRT-PCR was used to quantify specific tRNA abundances. Polysome profiling identified METTL1-dependent translational targets, and mitochondrial function was assessed by Seahorse Mito Stress testing. Results: METTL1 was significantly upregulated in PDAC (p < 0.001) and correlated with poor patient survival (p < 0.05). METTL1 knockout suppressed PDAC cell proliferation, migration, and tumor growth (p < 0.001) in vitro and in vivo. Mechanistically, METTL1 loss decreased the abundance of m7G-modified valine tRNAs, particularly Val-AAC, Val-CAC, and Val-TAC (all p < 0.001), leading to reduced translation of valine codon-enriched oxidative phosphorylation genes. Consequently, METTL1 deficiency impaired mitochondrial respiration and energy production. Consistently, valine tRNA expression was elevated in PDAC tissues (p < 0.01), and selective knockdown of these tRNAs inhibited proliferation and migration (p < 0.001) by disrupting mitochondrial function. Reintroduction of Val-AAC, Val-CAC, and Val-TAC restored growth (p < 0.001) and migratory capacity (p < 0.01) in METTL1-deficient cells. Conclusions: METTL1-mediated m7G modification of valine tRNAs promotes PDAC progression by reprogramming the translational landscape to support mitochondrial metabolism. This METTL1-tRNA-mitochondrial axis represents a novel metabolic vulnerability and a promising therapeutic target in pancreatic cancer. Citation Format: jiabei zhu, Qi Zhang, Rui Su, Qiuhui Pan, Ajay Goel. METTL1-mediated m7G modification of valine tRNAs drives pancreatic ductal adenocarcinoma progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 5962.

Hepatoblastoma (HB) is a rare but predominant liver cancer in children, with few treatment choices in advanced stages. YWHAE is closely related to several human diseases and acts as a molecular scaffold for malignant transformation. However, whether YWHAE promotes HB development remains unknown. Conducting RNA and m6A sequencing on HB tissues, we found that YWHAE was upregulated and modified by N6-methyadenosine. Functionally, YWHAE promoted proliferation and inhibited cell death in HB by in vitro and in vivo studies. Mechanistically, METTL3-dependent m6A modification activated YWHAE mRNA expression, and the m6A reader IGF2BP2 recognized and bound to the m6A site on YWHAE mRNA, thereby enhancing the mRNA stability of YWHAE. Interestingly, RNA sequencing revealed that YWHAE knockdown was involved in regulating ferroptosis of HB cells by mediating SLC7A11 expression. Moreover, knockdown of YWHAE significantly increased the levels of lipid ROS and peroxides in HB cells, promoting the susceptibility of HB cells to ferroptosis. In summary, these findings illuminated the role of YWHAE in HB progression and uncovered its relevance to ferroptosis as a new therapeutic target for HB.

Hepatoblastoma (HB) is the predominant hepatic malignancy among children. Despite therapeutic options for HB were gradually refined in recent years, patients with metastasis suffer from an unsatisfactory prognosis. Pyroptosis is a type of programmed and inflammatory necrosis. Neutrophils are crucial in innate immunity, which were shown to be associated with tumor progression. Our study strived to unravel the relationship between neutrophil-induced pyroptosis (NIP) and HB. The clinical and bulk RNA sequencing data of 38 patients with HB were obtained from Shanghai Children’s Medical Center. We established NIP score based on the LASSO regression. The single-cell RNA sequencing data (GSE186975) were used for for key genes identification, cellular communication, and differentiation trajectories of neutrophils. KEGG, GO, GSVA, and ssGSEA enrichment were used to analyze biological functions, including neutrophil extracellular traps (NETs), NOD-like receptors pathway, neutrophil activation, neutrophil-mediated cytotoxicity, and others. We constructed a NIP score based on the expression of three genes related to neutrophil and pyroptosis, namely ELANE, CASP1, and NOD2, which was positively correlated with a favorable prognosis of HB. Moreover, we clarified the function of ELANE in HB microenvironmwnt. Immunohistochemistry and transcriptome analysis unraveled a significant correlation between NETs and pyroptosis in HB, suggesting the key role of NETs-related neutrophils in inducing pyroptosis and prolonging survival. We also found upregulated tumor-promoting and immunosuppression-related pathways in the HB microenvironment. In addition, we clarified the growth trajectories and phenotypic changes of neutrophils in the immune microenvironment of HB, which can serve as potential targets for immunotherapy. The novel NIP score for patients with HB shows high predictive value for survival. Moreover, we identified biological function, cellular communication, and growth trajectories of neutrophils in HB. Our findings broaden insights into the treatment of HB.

Hepatoblastoma (HB) is the most common pediatric liver malignancy with an increasing incidence. However, the functional roles of 3D chromatin organization, epigenetic regulatory factors, and transcriptional reprogramming in HB pathogenesis remain poorly understood. ​Integrated multi-omics analyses of HB and matched non-tumor tissues were performed, including Hi-C, H3K27ac CUT Tag, ATAC-seq, and RNA-seq, to construct high-resolution 3D epigenomic maps and identify genes interacting with HB-specific super-enhancers (SEs). Functional assays of identified targets were conducted in cell lines and animal models. The regulatory mechanisms of SEs and upstream transcription factors (TFs) were investigated using CRISPRi-dCas9, 3C-qPCR, ChIP-qPCR, and luciferase reporter assays. Comprehensive analysis identified TRIB2 as an HB-specific SE-associated oncogene. Functionally, TRIB2 promoted cell proliferation and accelerated tumor growth both in vitro and in vivo. Patients with high TRIB2 expression exhibited advanced PRETEXT stage and metastasis. Mechanistically, TCF3 directly bound to both the TRIB2-SE and its promoter, promoting TRIB2 overexpression. Moreover, TRIB2 conferred resistance to ferroptosis by disrupting KEAP1-mediated ubiquitination of NRF2, thereby stabilizing NRF2 protein and enhancing antioxidant responses. The TCF3-TRIB2-NRF2 axis showed significant co-expression in HB tissues, effectively distinguished HB from normal liver tissues, and was associated with poorer overall survival. Our findings reveal that TCF3 and SE mediate TRIB2 overexpression to inhibit ferroptosis via the KEAP1-NRF2 pathway and drive HB pathogenesis, providing potential diagnostic and prognostic markers for HB.

Understanding the heterogeneity of epigenetic modifications within single cells is pivotal for unraveling the nature of the complexity of gene expression and cellular function. In this study, we have developed a strategy based on multichrome encoding and "AND" Boolean logic recognition for multiplexed, spatially resolved imaging of single-cell RNA epigenetic modifications, termed as PRoximity Exchange-assisted Encoding of Multichrome (PREEM). Through the implementation of this strategy, we can now map the expression and nuclear distribution of multiple site-specific RNA N6-methyladenosine (m6A) modifications at the single-molecule resolution level in single-cells, and reveal the previously unknown heterogeneity. Notably, we demonstrate how these patterns change after treatment with various drugs. Moreover, cyclic imaging with tailed DNA self-assembly further suggest the scalability and adaptability of PREEM's design. As an innovative epigenetic modification imaging tool, PREEM not only broadens the horizons of single-cell epigenetics research, enabling joint analysis of multiple targets beyond the limitations of imaging channels, but also reveals cell-to-cell variability, thereby enhancing our capacity to explore cellular functions.

Hepatoblastoma, the most aggressive childhood liver tumor, poses significant challenges due to limited knowledge of its pathogenesis, particularly in poorly differentiated advanced tumors where the prognosis is dismal. Single-cell sequencing provides an in-depth exploration at the single-cell level and offers a deep understanding of tumor heterogeneity. Herein, single-cell transcriptomics analysis is used to identify a unique malignant-hepatoblast (HB)-like cell subpopulation as the possible origin of poorly differentiated hepatoblastoma. These cells are associated with an unfavorable clinical prognosis in hepatoblastoma patients. The malignant-HB-like cell subpopulation generated insulin-like growth factor 2 (IGF2) to sustain stem-like features by promoting abnormal cholesterol accumulation via SREBF2. IGF2 also stimulated fibroblast 2 to secrete collagen 1, intensifying tumor malignancy via the collagen 1/integrin α1 signaling pathway. This suggests that targeting malignant HB-like cells by inhibiting IGF2-induced pathways can lead to promising treatments for hepatoblastoma. Additionally, serum IGF2 levels may serve as a diagnostic biomarker for advanced hepatoblastoma. In summary, these findings provide valuable insight into the genesis and malignancy of hepatoblastoma and a foundation for more effective diagnostic tools and therapeutic strategies for this challenging disease.

The current understanding of the relationship between circulating metabolites and neuroblastoma (NB) risk is insufficient. Herein, bidirectional Mendelian randomization (MR) studies were utilized to examine the potential causal associations of metabolites and the risk of NB. Significant single-nucleotide polymorphisms (SNPs) associated with circulating metabolites were obtained from the genome-wide association study (GWAS) in the European population (N = 7824). Summary statistics for NB were aggregated from prior GWAS studies encompassing 1,627 patients and 3,254 cancer-free children. The causality was assessed primarily by inverse-variance weighted (IVW), along with MR-Egger regression, weighted median estimator, and weighted mode method. Among the 486 metabolites analyzed, a genetically determined elevation in blood butyrylcarnitine concentration (log10) was significantly correlated with a 51.5

With the development of omics technology, multiple omics techniques have greatly expanded the effective data volume and shown great value in accurately diagnosing major diseases. However, the extremely limited clinical samples and a wide variety of detection biomarkers raise the key technical issue of how to use a small number of samples to achieve multiple biomarker detection. DNA self-assembly (DSA) encoding technology is a controllable technique that can achieve precise regulation of detection signals by combining fluorescent molecular markers based on the high programmability of DNA and the high predictability of the assembled products, playing an important role in the field of target multiplex analysis. Here, we provide a comprehensive overview of DSA encoding in spatial multi-omics imaging, further point out current problems in multiplex analysis, and put forward future research directions.

N6-Methyladenosine (m6A) is one of the most abundant RNA modifications that occur in eukaryotes. The relationship between m6A methylation and DNA damage repair (DDR) is still unclear. As an important chaperone protein of m6A methyltransferase, the role of Wilm’s tumor–associated protein (WTAP) in DDR has not been studied in detail. We identified that WTAP was markedly upregulated in HCC cells with DNA damage induction. Further investigations revealed that WTAP was engaged in DDR and WTAP knockdown resulted in a significant decrease of the stability of the DDR-related transcription factor Forkhead Box M1 (FOXM1) mRNA, eventually preventing DDR. WTAP deficiency inhibited cell growth both in vivo and in vitro. Moreover, WTAP silencing sensitized HCC cells to cisplatin, a well-known DNA damage agent. Altogether, our findings demonstrate that WTAP is the dominant m6A -related modulator upon DNA damage and functions by stabilizing FOXM1. In addition, WTAP deficiency saliently sensitized HCC cells to cisplatin both in vitro and in vivo.

BACKGROUND:The relationship between sex hormone-binding globulin (SHBG) levels and the risk of adverse pregnancy outcomes (APOs) remains controversial. A two-sample Mendelian randomization (MR) study was performed to clarify the causality of SHBG on the risk of APO. METHODS:Significant single-nucleotide polymorphisms (SNPs) associated with SHBG levels were obtained from the genome-wide association study (GWAS) in the European population. Summary statistics of the number of spontaneous miscarriages, preeclampsia, gestational diabetes mellitus (GDM), intrahepatic cholestasis of pregnancy (ICP), and female infertility were utilized as the outcome. The causality was examined primarily by inverse-variance weighted (IVW), along with MR-Egger regression, weighted median estimator, and weighted mode method. RESULTS:Based on the IVW model, every genetically predicted standard deviation (SD) increase in SHBG levels was causally associated with 0.023 SDs decrease of the number of spontaneous miscarriages (Beta ± SE: -0.023 ± 0.010, p = 0.018), 11.3% decrease of the risk of preeclampsia (OR = 0.887, 95% CI: 0.806-0.977, p = 0.015), 17% decrease of the risk of GDM (OR = 0.830, 95% CI: 0.753-0.914, p = 0.000), 23.6% decrease of the risk of ICP (OR = 0.764, 95% CI: 0.584-0.999, p = 0.049), and 14% decrease of the risk of infertility (OR = 0.860, 95% CI: 0.777-0.951, p = 0.003). CONCLUSION:Our study indicated that the increased levels of SHBG could significantly reduce the risk of APO. SHBG may be helpful as the indicator for preconception risk assessment and pregnancy risk monitoring. These findings are limited to European and require further validation in diverse populations.

Background: Neuroblastoma (NB) is the most common extracranial malignant solid tumor in children, accounting for >15 % of cancer-related deaths in children. We analyzed the epidemiological statistical indicators of neuroblastoma and other peripheral nervous system tumors patients from 1990 to 2021 in Global Burden of Disease (GBD) 2021 database, aiming to provide valuable insights for public health interventions and clinical practices. Methods: Based on the GBD 2021 database, this study analyzed the incidence, mortality, prevalence, and Disability-Adjusted Life-Years (DALYs) of neuroblastoma and other peripheral nervous system tumors from 1990 to 2021, stratified by sociodemographic development index (SDI) and geographic regions. Cross-country inequalities analysis was conducted to quantify the SDI-related inequality of disease burden across countries. In addition, the average annual percentage change (AAPC) and Age-Period-Cohort (APC) model were used to evaluate the trend of disease burden, while the global burden of disease to 2035 was predicted by Bayesian Age-Period-Cohort (BAPC) model. Findings: This study reported the disease burden of neuroblastoma and other peripheral nervous system tumors in GBD 2021 database for the first time. Globally, the incidence and mortality of neuroblastoma have increased year by year from 1990 to 2021, especially in regions with low SDI, such as South Asia and sub-Saharan Africa, where the burden of disease has increased significantly. Regions with high SDI, such as North America and Western Europe, have seen a reduction in disease burden due to higher levels of medical care and earlier diagnosis. The age distribution shows that children under 5 years of age are mainly affected, especially in low- and middle-income areas. In addition, the incidence is slightly higher in men than in women. The BAPC model predicts that the global incidence, mortality, and DALYs of neuroblastoma will continue to increase until 2035. Interpretation: Significant regional and population variation in neuroblastoma and other peripheral nervous system tumors worldwide, with a particularly high disease burden in low SDI areas with limited medical resources. This trend highlights the urgent need for global public health interventions and resource allocation, particularly in low-income countries. Future research should focus on improving early diagnosis, risk stratification and target therapy in order to reduce the global burden of disease and improve patients’ prognosis. Funding: This study was supported by National Natural Science Foundation of China (No. 82293662, No 82172357 and No 81930066), Key project of Shanghai ''Science and Technology Innovation Action Plan (22JC1402304) and Research fund of Shanghai Municipal Health Bureau (No. 2019cxjq03).

Hepatoblastoma (HB), the most common pediatric hepatic malignancy, exhibits an increasing incidence. Metabolism reprogramming represents a pivotal hallmark in the oncogenic transformation process, with glutamine emerging as a critical energy source for neoplastic cells, rivaling glucose. However, the mechanism by which glutamine is involved in the development of HB remains unclear. Our study identified glutamine metabolism as a crucial factor in the development of HB. The key enzyme of glutamine metabolism, kidney-type glutaminase (GLS1), is activated in HB and regulates cell proliferation. Mechanistically, the GLS1 subtype KGA, utilizing glutamate derived from glutaminolysis, enhances glutathione (GSH) synthesis, which in turn inhibits ferroptosis in HB cells. Importantly, the Thr563 residue of KGA undergoes O-GlcNAcylation, enhancing enzyme activity and stability, accelerating glutaminolysis, and promoting the proliferation of HB. This study demonstrated that enhanced glutaminolysis, driven by GLS1, is crucial for the development of HB by inhibiting ferroptosis. The O-GlcNAcylation of KGA isoform ensures its stability and glutaminase function in HB cells, which can serve as a promising therapeutic target for KGA-mediated glutaminolysis in HB.

BACKGROUND:Hepatoblastoma is the most prevalent liver cancer in children. Immunotherapy targeting immune checkpoint molecules has become pivotal in various cancer treatments. However, the clinical significance of immune checkpoint ligands in hepatoblastoma remains largely unclear due to various challenges. This study sought to first characterize the expression profile of the immune checkpoint ligand CD276 in hepatoblastoma and assess its potential as a predictor of malignant characteristics and regulator of neutrophil infiltration. METHODS:Univariable and multivariable logistic regression analyses were performed to evaluate the clinical significance of immune checkpoint ligands in the bulk RNA-seq dataset and develop a novel predictive model for malignancy. Furthermore, single-cell RNA sequencing (scRNA-seq), immunohistochemistry (IHC), deconvolution analysis, and correlation analysis were employed to characterize the expression pattern of CD276 and explore its influence on the tumor immune microenvironment. RESULTS:The bulk RNA-seq analysis revealed CD276 transcript levels were significantly elevated in hepatoblastoma tissues, especially in patients with more aggressive malignant phenotypes. Furthermore, we developed a predictive model based on a risk score and constructed a user-friendly nomogram to predict patient metastasis by integrating CD276 levels with clinical features. Both scRNA-seq and multiplex immunohistochemistry (mIHC) analyses confirmed that CD276 is highly expressed, predominantly in cancer-associated fibroblasts. Our results also demonstrated that CD276 levels correlate with immune infiltration in hepatoblastoma, and that CD276 regulates CXCL2 to modulate neutrophil infiltration, suggesting a potential mechanism underlying the role of CD276 in hepatoblastoma malignancy. In addition, we validated that CD276 in cancer-associated fibroblasts promotes hepatoblastoma growth in mice. CONCLUSIONS:Our findings highlight the critical role of CD276 in hepatoblastoma malignancy, potentially providing novel insights and therapeutic targets for the development of combinatorial immunotherapy strategies for this disease.