Primary liver cancer remains a significant global public health challenge, characterized by persistently high incidence and mortality. This review synthesizes current epidemiological data to analyze trends and etiological shifts, with particular emphasis on China, which bears over 40% of the global burden. Findings highlight a transition in dominant risk factors from viral hepatitis to metabolic dysfunction-associated steatotic liver disease (MASLD), alongside persistent threats from aflatoxin exposure and lifestyle behaviors. Evidence-based prevention strategies, including universal hepatitis B virus (HBV) vaccination, antiviral therapy expansion, aflatoxin control, and early metabolic intervention, are critical to reducing disease burden. The integration of artificial intelligence into screening and management represents a promising advancement. A multi-faceted approach combining vaccination, surveillance, lifestyle modification, and technological innovation is essential for effective global liver cancer control.

Intrahepatic cholangiocarcinoma (iCCA) is the second most prevalent liver cancer with a high mortality and recurrence rate, and remains a poorly understood disease. The tumor margin, as the transition zone between normal tissue and tumor, was not appreciated before. We performed mass cytometry (cytometry by time of flight, CyTOF) on 30 samples from iCCA tumor, paratumor and margin tissue. We found that the number of CD4+ central memory T cells (CD4+ Tcm) increased in the margin zone. Single cell RNA-seq (scRNA-seq) further discovered that CD4+ Tcm cells enriched in the margin zone of iCCA exhibited upregulated Annexin A1 (ANXA1) expression. Fibroblasts recruited CD4+ ANXA1+ Tcm cells via Chemokine (C-C motif) ligand 19 (CCL19)-Chemokine (C-C motif) receptor 7 (CCR7) signaling pathway. The molecular characteristics of CD4+ ANXA1+ Tcm in iCCA were characterized, and the interactions between these T cells and other cells were determined. Patients with enriched CD4+ ANXA1+ Tcm cells in the tumor margin exhibited a worse prognosis. Specifically, CD4+ ANXA1+ Tcm cells could recruit macrophages to the tumor margin and regulate the macrophage polarization via the ANXA1-Formyl Peptide Receptor 1 (FPR1) signaling axis. CD4+ ANXA1+ Tcm cell-activated macrophages could enhance the invasion and proliferation of tumor cells by exerting different cytokines. In conclusion, our study systematically characterized the features and distribution of CD4+ ANXA1+ Tcm cells in the margin zone of iCCA. We investigated the potential mechanisms by which CD4+ ANXA1+ Tcm cells affected tumor progression, and provided novel understanding of the function of these CD4+ Tcm cells in iCCA.

Abstract Background Tumor heterogeneity presents a formidable challenge in understanding the mechanisms driving tumor progression and metastasis. The heterogeneity of hepatocellular carcinoma (HCC) in cellular level is not clear. Methods Integration analysis of single-cell RNA sequencing data and spatial transcriptomics data was performed. Multiple methods were applied to investigate the subtype of HCC tumor cells. The functional characteristics, translation factors, clinical implications and microenvironment associations of different subtypes of tumor cells were analyzed. The interaction of subtype and fibroblasts were analyzed. Results We established a heterogeneity landscape of HCC malignant cells by integrated 52 single-cell RNA sequencing data and 5 spatial transcriptomics data. We identified three subtypes in tumor cells, including ARG1+ metabolism subtype (Metab-subtype), TOP2A+ proliferation phenotype (Prol-phenotype), and S100A6+ pro-metastatic subtype (EMT-subtype). Enrichment analysis found that the three subtypes harbored different features, that is metabolism, proliferating, and epithelial-mesenchymal transition. Trajectory analysis revealed that both Metab-subtype and EMT-subtype originated from the Prol-phenotype. Translation factor analysis found that EMT-subtype showed exclusive activation of SMAD3 and TGF-β signaling pathway. HCC dominated by EMT-subtype cells harbored an unfavorable prognosis and a deserted microenvironment. We uncovered a positive loop between tumor cells and fibroblasts mediated by SPP1-CD44 and CCN2/TGF-β-TGFBR1 interaction pairs. Inhibiting CCN2 disrupted the loop, mitigated the transformation to EMT-subtype, and suppressed metastasis. Conclusion By establishing a heterogeneity landscape of malignant cells, we identified a three-subtype classification in HCC. Among them, S100A6+ tumor cells play a crucial role in metastasis. Targeting the feedback loop between tumor cells and fibroblasts is a promising anti-metastatic strategy.

Abstract Gut microbiome alterations are increasingly associated with hepatocellular carcinoma (HCC), highlighting the gut–liver axis as a key contributor to tumor progression and prognosis. Taxon‐based HCC microbiome studies have shown limited reproducibility because they are affected by database dependency, taxonomic ambiguity, and overlooked ecological interactions. The Two Competing Guilds (TCG) model, based on stable gut microbiome interactions, provides a structurally grounded framework for robust, generalizable biomarkers. Using shotgun metagenomic data from a newly recruited cohort of 120 surgically resectable HCC cases and 76 benign liver tumor controls, we constructed co‐abundance networks to identify stably correlated genome pairs and assembled a hepatic cancer‐TCG (HCC‐TCG) model composed of 142 genomes. Functionally, one Guild had more genes for butyrate production from carbohydrate fermentation while the other Guild was enriched in genes for virulence factors and antibiotic resistance, highlighting its potential proinflammatory roles. Classifiers trained on the abundance profiles of HCC‐TCG genomes successfully distinguished HCC from benign liver tumors (area under the receiver operating characteristic, AUROC = 0.70) and from colorectal liver metastases (CRLM) (AUROC = 0.78). In an external validation cohort, the model further discriminated against HCC from intrahepatic cholangiocarcinoma (iCCA) (AUROC = 0.72), and from healthy controls (AUROC = 0.79–0.85), demonstrating its broad applicability for tumor stratification across clinical contexts. Moreover, HCC‐TCG profiles predicted post‐resection recurrence risk and response to adjuvant therapies (AUROC up to 0.83). Importantly, external validation in two independent cohorts of advanced HCC patients treated with PD‐1/PD‐L1 inhibitors demonstrated consistent predictive performance (AUROC = 0.64–0.73), confirming the model's generalizability in nonsurgical and immunotherapy contexts. This genome‐specific, ecologically structured, and database‐independent framework identifies a conserved Guild‐based microbiome signature for HCC. Our findings demonstrate that a fixed genome‐resolved ecological structure retains transferable discriminatory signal across clinical contexts. The HCC‐TCG framework provides a genome‐specific, interaction‐based foundation for future development of non‐invasive microbiome stratification strategies requiring prospective validation.

e16291 Background: Clinical management of hepatocellular carcinoma (HCC) is becoming increasingly complex, encompassing curative resection for early-stage disease and multimodal treatment strategies across disease stages. However, preoperative risk stratification and timely evaluation of treatment response remain challenging due to the lack of reliable blood-based tools. Tumor-agnostic circulating tumor DNA (ctDNA) methylation profiling may help address these unmet clinical needs by enabling real-time, noninvasive assessment of tumor biology and therapeutic efficacy. Methods: We prospectively enrolled 174 HCC patients undergoing curative hepatectomy and collected plasma samples within 24 hours prior to surgery (Ts). In parallel, an independent neoadjuvant therapy cohort of 27 HCC patients treated with immune checkpoint inhibitor-based systemic therapy was included, in which plasma samples were obtained at baseline before treatment initiation (Tb) and prior to surgery (Ts). All plasma samples were assayed using the GutSeer. A tumor-agnostic machine learning model was developed based on 342 HCC patients and 1,415 healthy subjects to generate a methylation-based cancer score (MCS) for prognostic stratification and pathological response assessment. Results: The hepatectomy cohort was followed for a median of 26.3 months with 47 recurrences. Preoperative Ts MCS was significantly associated with advanced tumor stage, larger tumor size, and vascular invasion, indicating its ability to reflect tumor burden and underlying biological aggressiveness. Based on Ts MCS, patients were stratified into low-risk (n = 45) and high-risk (n = 129) groups, yielding a hazard ratio (HR) of 9.36 (95%CI: 2.27–38.59; p < 0.001) and a negative predictive value of 95.6%. Multivariate analysis confirmed Ts MCS as an independent predictor of recurrence (HR = 5.09, 95%CI: 1.18-21.92; p < 0.05). In the neoadjuvant therapy cohort, MCS decreased significantly from Tb to Ts (p < 0.001). The change of MCS (Tb-Ts) was strongly negatively correlated with the proportion of residual viable tumor (R = -0.70, p < 0.001), indicating MCS changes closely mirror pathological response. Consistently, MCS change accurately discriminated major pathological response (MPR, ≤50% viable tumor cells) from non-MPR (AUC = 0.859), outperforming changes of alpha-fetoprotein (AUC = 0.731) and des-gamma-carboxy prothrombin (AUC = 0.686). Conclusions: This study demonstrates that tumor-agnostic ctDNA methylation profiling enables effective preoperative risk stratification in resectable HCC. Furthermore, dynamic changes in MCS provide a precise, noninvasive biomarker for assessing pathological response to systemic therapy. These findings highlight the broad clinical utility of methylation-based liquid biopsy approaches across surgical and systemic treatment settings in HCC. Clinical trial information: NCT06178809 .

e16223 Background: Hepatocellular carcinoma (HCC) remains challenging to manage once unresectable. Although atezolizumab plus bevacizumab is standard first-line therapy, its potential to enable conversion to curative resection is not well defined. We evaluated the efficacy and safety of atezolizumab plus bevacizumab combined with on-demand transarterial chemoembolization (TACE) in a real-world cohort, with a focus on surgical conversion. Methods: We conducted a multicenter real-world study of 121 patients with unresectable HCC treated with atezolizumab plus bevacizumab ± TACE. The primary endpoint was the objective response rate (ORR), and secondary endpoints included progression-free survival (PFS), overall survival (OS), disease control rate, surgical conversion rates and safety. Results: Patients treated in the first-line setting achieved better outcomes than those receiving second-line therapy (ORR 47.3%, 53/112, vs 22.2%, 2/9), with longer median PFS (19.0 vs 8.8 months) and OS (35.3 vs 13.0 months). In the first-line cohort, patients receiving atezolizumab plus bevacizumab with TACE showed higher ORR (50.5%, 48/95, vs 29.4%, 5/17), longer median PFS (20.9 vs 8.0 months) and OS (35.3 vs 25.6 months) compared with those without TACE, with outcomes numerically exceeding historical GO30140 and IMbrave150 benchmarks. Successful R0 resection was achieved in 33.7% (32/95) of patients in the with-TACE group versus 5.9% (1/17) in the without-TACE group. Among resected patients, pCR occurred in 34.4% (11/32) and MPR in 68.8% (22/32; MPR, < 50% viable tumor cells). Among patients achieving MPR, RECIST responses included CR (n = 3), PR (n = 10), and SD (n = 9), indicating incomplete concordance between pathological and radiological assessments. A single tumor and higher CD8 + T-cell infiltration in baseline biopsy specimens were associated with a more favourable ORR. We further identified ECOG performance status, lower tumor burden, and pathological features of baseline biopsy specimens (higher CD4 + and CD8 + T-cell infiltration) as key factors associated with successful conversion to surgery. Treatment-related adverse events (TRAEs) occurred in 91.6% and 88.2% of patients in the with- and without-TACE groups, with grade 3/4 events in 54.7% and 52.9%, respectively; no grade 5 events or postoperative complications were observed. Conclusions: In this multicenter real-world cohort, on-demand TACE combined with atezolizumab plus bevacizumab enabled deep tumor responses and curative-intent resection in one-third of initially unresectable HCC patients. Baseline immune infiltration identified candidates most likely to benefit from conversion. These findings support a conversion-oriented strategy integrating locoregional and systemic therapy to bridge unresectable HCC toward surgical cure.

Circular RNAs (circRNAs) are covalently closed, single-stranded RNAs generated via backsplicing. They are highly stable and evolutionarily conserved, making them promising candidates for cancer therapy and diagnosis. CircRNAs regulate cancer progression by modulating genome instability, angiogenesis, metastasis, stemness, and chemoresistance. They do so through mechanisms including microRNA (miRNA) sponging, protein interaction, translational templating, and transcription/translation regulation. CircRNAs play a critical role in cancer immunotherapy. They modulate immune checkpoint blockade (ICB) responses and cytokine secretion to reshape the tumor immune microenvironment (TME). CircRNAs also serve as stable platforms for neoantigen-based cancer vaccines and improve in vivo chimeric antigen receptor T cell (CAR-T) therapy by replacing unstable linear mRNA. Additionally, circRNAs are potential noninvasive biomarkers due to their abundance in body fluids and differential tumor-normal expression. Despite challenges such as unclear regulatory networks, off-target effects, and inefficient delivery, this review systematically summarizes the biogenesis of circRNAs, their functional mechanisms, their roles in cancer progression, and their applications in cancer immunotherapy. The review also highlights their utility as biomarkers and future translational directions, providing a focused overview of their potential to advance cancer immunotherapy.

Idiopathic pulmonary fibrosis (IPF) is a progressive and fatal interstitial lung disease with limited therapeutic options, highlighting the need for new molecular targets and effective antifibrotic agents. In this study, we investigated the role of B-cell lymphoma 9 (BCL9) in IPF and evaluated the antifibrotic potential of ZD-4009, a novel small-molecule inhibitor targeting BCL9. BCL9 was upregulated in fibrotic lung tissues, suggesting a possible association between BCL9 and pulmonary fibrosis progression. ZD-4009 showed high-affinity binding to β-catenin (pKD = 7.36 ± 0.01, KD = 44.72 nM). In vitro, ZD-4009 inhibited fibroblast activation, suppressed extracellular matrix deposition, and reduced the expression of fibrosis-associated markers, including COL1A1 and α-SMA. Mechanistically, ZD-4009 suppressed HIF-1α-associated glycolytic reprogramming and inhibited TGF-β/SMAD signaling, suggesting that its antifibrotic effects are associated with coordinated regulation of metabolic and profibrotic pathways. In vivo, ZD-4009 treatment was associated with alleviated bleomycin-induced pulmonary fibrosis, as reflected by improved survival from 55% in the bleomycin group to 82% in the bleomycin + ZD-4009 group, reduced relative collagen content from 61% to 48%, and ameliorated micro-CT-assessed fibrotic changes. In the tested model, ZD-4009 showed favorable antifibrotic effects compared with nintedanib, while liposome-encapsulated ZD-4009 reduced treatment-related toxicity. Together, these findings nominate ZD-4009 as a promising antifibrotic candidate and highlight BCL9 as a potential therapeutic target for IPF.

BACKGROUND:No neoadjuvant treatment has been considered to be standard therapy for patients with resectable intrahepatic cholangiocarcinoma with high-risk factors for recurrence. The GOLP regimen (gemcitabine-oxaliplatin, lenvatinib, and an anti-programmed death 1 antibody) has shown promising efficacy with a manageable safety profile in advanced intrahepatic cholangiocarcinoma and biliary tract cancer. METHODS:In a phase 2-3 trial, we randomly assigned, in a 1:1 ratio, patients with resectable high-risk intrahepatic cholangiocarcinoma to the neoadjuvant group (intravenous gemcitabine-oxaliplatin plus toripalimab every 3 weeks for three cycles and oral lenvatinib once daily for 9 weeks, followed by curative resection) or the control group (curative resection and no neoadjuvant treatment). All patients received adjuvant capecitabine for eight cycles after surgery. The primary end point was event-free survival. Secondary end points included overall survival and safety. RESULTS:A total of 178 patients underwent randomization (88 patients to the neoadjuvant group and 90 to the control group). At the interim analysis at a median follow-up of 16.9 months, the median event-free survival was significantly longer in the neoadjuvant group (18.0 months; 95% confidence interval [CI], 13.8 to 27.6) than in the control group (8.7 months; 95% CI, 7.2 to 12.4) (P<0.001). Overall survival at 24 months was 79% (95% CI, 70 to 90) in the neoadjuvant group and 61% (95% CI, 50 to 75) in the control group (hazard ratio for death, 0.43; 95% CI, 0.23 to 0.79; P = 0.005, which did not meet the significance criterion [two-sided alpha, 0.0019]). Across all treatment phases, adverse events occurred in 97% of the patients in the neoadjuvant group and in 70% of those in the control group. During the neoadjuvant phase, adverse events of grade 3 or higher occurred in 28% of the patients, and treatment-related adverse events of grade 3 or higher in 26%. No treatment-related adverse event led to death. CONCLUSIONS:Neoadjuvant GOLP led to significantly longer event-free survival than control therapy, with mainly low-grade adverse events, among patients with resectable high-risk intrahepatic cholangiocarcinoma. (Funded by the Clinical Research Plan of Shanghai Hospital Development Center and others; ZSAB-neoGOLP ClinicalTrials.gov number, NCT04669496.).

Liver cancer, particularly hepatocellular carcinoma (HCC), poses a severe global public health threat owing to its high incidence, frequent late-stage diagnosis, and poor 5-year survival rate. Conventional approaches to liver cancer diagnosis and treatment are limited by their reliance on subjective physician experience, uniform and undifferentiated treatment strategies, and imprecise prognostic assessment. This review synthesizes studies published between 2019 and 2025 on the application of multi-modal data in liver cancer care, including computed tomography (CT), magnetic resonance imaging (MRI), pathology, and multi-omics data. We explore the utility of single-modal data analysis including the role of CT or MRI in enhancing diagnostic accuracy and the application of pathological data. Subsequently, the review focuses on multi-modal data fusion strategies, including feature-level, decision-level, and modal-level fusion, which collectively support precision diagnosis, personalized treatment recommendation, and accurate prognosis prediction in clinical practice. Additionally, it addresses critical challenges such as data heterogeneity and low physician acceptance of integrated data-driven tools, while outlining future directions including the development of standardized multi-modal data ecosystems. This review highlights multi-modal data as a core driver of precision liver cancer care, with the objective of accelerating its translation into routine clinical practice.

Abstract Background and aims Existing imaging and serum‐marker assays miss many early liver cancers, especially in high‐risk chronic liver disease carriers. We aimed to create a highly accurate, non‐invasive, methylation‐based liquid biopsy for early detection. Methods We used a comprehensive, multi‐platform, multi‐cohort strategy for marker discovery, starting with methylation profiling of hepatocellular carcinoma samples from TCGA and in‐house cohorts. From 30 initial candidates, nine highly liver‐specific methylation markers were shortlisted, and three optimal cfDNA markers (RNF135, CHFR, PAX5) were selected to develop a robust diagnostic model, tuned in a training set (N = 280) and locked in an internal testing set (N = 124). The model was then validated in a prospective, large‐scale trial conducted at four geographically distinct Chinese centres. Results The clinical trial included 1097 participants from two groups, (i) a diagnosing group (N = 646) that prospectively enrolled individuals without prior diagnostic results and represented a real‐world high‐risk population, and (ii) a diagnosed group recruited after pathology confirmation. Overall, the model achieved 94.43% (95% confidence interval, 92.12–96.09%) sensitivity and 95.16% (92.78–96.78%) specificity for liver cancer, with stage‐I sensitivity of 93.10% (89.78–95.40%). Within the diagnosing group, overall sensitivity was 93.99% (91.28–95.90%), and for the 267 stage‐I cases, it reached 92.88% (89.15–95.39%). As for specificity, it remained high across confounders: 92.78% (85.84–96.46%) in cirrhosis, 91.74% (85.46–95.45%) in other‐cancer interference samples. Besides, the model outperformed the traditional liver cancer biomarker AFP and showed changes in methylation signals before and after surgery, suggesting a possible role in perioperative monitoring. Each centre independently reported sensitivities and specificities exceeding 90%, demonstrating robust geographic performance. Conclusions Using a systematic marker‐discovery pipeline and a multi‐centre prospective cohort, we developed a methylation‐based liquid biopsy that reliably detects early liver cancer in high‐risk populations. Clinical trial number Chictr.org identifier: ChiCTR2400092883. Key points Three cfDNA methylation markers, RNF135, CHFR and PAX5, were identified for liver cancer liquid biopsy. A three‐marker diagnostic model based on qMSP was established for highly accurate non‐invasive detection of liver cancer. The LC‐HMC model achieved 94.43% sensitivity and 95.16% specificity in the clinical trial. The model detected stage‐I liver cancer with a sensitivity of 93.10%.

BACKGROUND:Hepatocellular carcinoma (HCC) remains a leading cause of cancer mortality worldwide. While immune checkpoint blockade has revolutionized the treatment of many cancers, responses in HCC remain limited. Robust functional platforms capable of predicting individual responses to immunotherapy are urgently needed. In this study, we developed a patient-derived organotypic tumor spheroid (PDOTS) model that preserves the heterogeneity and immune microenvironment of HCC, enabling rapid and reliable assessment of targeted and immunotherapeutic responses. METHODS:Tumor tissues from 30 HCC patients were processed using a "Five-Point Clock" sampling method and cultured within a three-dimensional microfluidic chip supplemented with IL-2 and CD3/28 activator to maintain tumor-infiltrating lymphocyte activity. The genomic, immune, and histopathological fidelity of PDOTS relative to parental tumors was evaluated. Drug responses were assessed ex vivo and validated in matched patient-derived xenograft (PDX) models. Transcriptomic profiling was subsequently performed to identify gene signatures associated with treatment sensitivity and to construct a transcriptomic predictive model. RESULTS:The PDOTS retained ≥60% viability over 7 days and faithfully maintained the genomic, immune, and histopathological profiles of parental tumors. Functionally, PDOTS exhibited immune-dependent responses to PD-1 blockade and predicted treatment responses with 80% concordance in matched PDX models. Transcriptomic profiling revealed distinct metabolic and immune signatures in sensitive and resistant tumors, which were used to derive the "Organoid Killing Index (OKI)". The OKI gene-derived index, a composite index integrating the enrichment scores of gene signatures associated with the OKI, strongly correlated (R=0.829, p<0.001) and predicted clinical outcomes in an external patient cohort treated with atezolizumab plus bevacizumab. CONCLUSIONS:These findings establish PDOTS as an immune-competent ex vivo platform for functional precision oncology and support the integration of functional testing with transcriptomic prediction to guide individualized immunotherapy in HCC.

Tumor-associated macrophages (TAMs) are key regulators of the metastatic immune microenvironment, yet the specific TAM subsets that drive immune suppression and tumor progression in colorectal cancer liver metastasis (CRLM) remain poorly defined. Here, we integrated CyTOF, single-cell and spatial transcriptomics, bulk RNA sequencing, and lipidomics to identify a distinct population of lipid-laden, immunosuppressive TREM2+ TAMs enriched in CRLM. These cells exhibited high expression of lipid metabolism-related genes, including APOE, LIPA, and GPNMB, and accumulated abundant intracellular lipid droplets. Spatial analyses revealed their preferential localization at the invasive margins and within intratumoral colonic lumen-like structures-regions characterized by the buildup of APOE protein, mucinous material, and apoptotic tumor debris. Transcriptional analyses suggest these macrophages follow a Kupffer cell-related differentiation trajectory and acquire an immunoregulatory phenotype via uptake of tumor-derived lipids. Functionally, TREM2+ TAMs produced leukotrienes via the ALOX5/ALOX5AP pathway, which in turn sustained chronic inflammatory signaling. This inflammatory milieu potentiated neutrophil recruitment and fostered tumor cell stemness, thereby reinforcing an immunosuppressive metastatic niche and correlating with poor patient prognosis. In vivo murine depletion model and ex vivo organotypic tumor models confirmed that either selective ablation of TREM2+ TAMs or pharmacological inhibition of leukotriene synthesis alleviated immunosuppression and potentiated the efficacy of anti-PD-1 therapy. Our study defines a conserved lipid-associated TREM2+ TAM population as an essential contributor of immune evasion and microenvironment remodeling in liver metastasis, and suggest it as a potential therapeutic target in metastatic colorectal cancer.

4162 Background: Immune checkpoint inhibitor (ICI) based conversion therapy is increasingly utilized in the management of initially unresectable hepatocellular carcinoma (HCC). However, the clinical significance of immune-related pathologic response (irPR) features remain poorly understood in HCC. Methods: We enrolled 217 HCC patients undergoing ICI-based conversion therapy followed by curative-intent resection and assigned them to discovery and validation cohorts (2:1). Thirteen immune-related pathological response (irPR) features—tumor-infiltrating lymphocytes, tertiary lymphoid structures, lymphoid aggregates, plasma cell infiltration, granulomas, neutrophils, foamy macrophages, cholesterol clefts, hemosiderin-laden macrophages, giant cells, neovascularization, necrosis, and mature fibrosis—were assessed on H&E-stained tumor bed sections. To explore differences in the tumor microenvironment, bulk mRNA sequencing was performed on FFPE samples from patients in the discovery cohort. Results: In the discovery cohort, four irPR features (tumor-infiltrating lymphocytes, lymphoid aggregates, neutrophils, and foamy macrophages) were integrated to construct the immunotherapeutic response score (ITRS), classifying patients as ITRSlow (0–1) or ITRShigh (2–4). ITRS low patients showed significantly improved OS (HR = 4.17; 95% CI 1.64–10.60; P = 0.003) and RFS (HR = 2.07; 95% CI 1.32–3.23; P = 0.001), with longer median RFS (32.3 vs 8.5 months). ITRS remained independently associated with OS (HR = 2.81; 95% CI 1.06–7.43; P = 0.038) and RFS (HR = 2.10; 95% CI 1.32–3.33; P = 0.002) and was validated in an independent cohort. FFPE transcriptomic profiling (n = 41) showed that ITRS low tumors were immune-inflamed, with enrichment of immune-related GO/KEGG pathways and globally higher immune/stromal infiltration (higher ESTIMATE immune/stromal scores, lower tumor purity; all P < 0.001). Unsupervised clustering identified hot/cold states, with hot tumors enriched in ITRS low (65.4% vs 13.3%, P = 0.003); CD8 + T-cell infiltration was higher across MCPcounter/CIBERSORT/TIMER and was accompanied by stronger antigen presentation (MHC-II gene sets) and increased CD8 + T-cell activation/cytotoxicity as well as exhaustion signatures (all P < 0.05), supporting an immune-active microenvironment linked to favorable outcomes. Conclusions: An irPR feature–based ITRS robustly predicts both OS and RFS in HCC patients following ICT. The ITRS is readily applicable in routine diagnostic pathology practice and provides novel insights into the biological mechanisms underlying immunotherapy resistance.

BACKGROUND:Surgical site infection (SSI) is a serious and potentially lethal complication following liver surgery, which is commonly associated with nosocomial bacteria such as Klebsiella pneumoniae, Acinetobacter baumannii, Enterococcus faecalis, and Enterococcus faecium. To address the critical need for rapid on-site diagnosis of these infections in complex abdominal drainage fluid (ABDF), we developed the CTL-SSI-LAMP platform, a novel point-of-care system that uniquely integrates Chelex-100 thermal lysis (CTL) DNA extraction with lyophilized loop-mediated isothermal amplification (LAMP). RESULTS:The optimized CTL method, which utilizes Chelex-100 resin to adsorb inhibitors and thermal treatment for efficient cell lysis, allowed for rapid DNA extraction from ABDF samples within 15 min using minimal equipment. Following optimization, the analytical sensitivity of the lyophilized SSI-LAMP assays was ≤ 103 CFU/mL with satisfactory analytical specificity, which was validated through both simulated and real ABDF samples. The entire detection process takes approximately 1 h to be completed without requiring specialized laboratory equipment. In a clinical cohort, the clinical sensitivity of our platform for these four SSI-related bacteria was 100%, while the clinical specificity was over 97%, with a negative predictive value of 100% and an overall accuracy exceeding 98%. SIGNIFICANCE:The CTL-SSI-LAMP platform provides a rapid, portable, and culture-independent method for detecting major SSI-related bacteria directly from postoperative ABDF. By combining robust sample pretreatment with lyophilized reagents, this platform enables timely, targeted antimicrobial therapy and is highly suitable for point-of-care application in intensive care units, operating rooms, and resource-limited clinical settings.

e16245 Background: KRAS mutations are common in colorectal cancer and associated with poor outcomes after colorectal liver metastases (CRLM) resection. However, the relationship between specific KRAS subtypes and survival/recurrence in CRLM patients post-resection is not well-defined. Methods: We retrospectively analyzed 1389 CRLM patients undergoing curative-intent hepatectomy (2020–2022). KRAS variants were determined by targeted sequencing with Sanger confirmation. OS and PFS were analyzed by Kaplan–Meier/log-rank tests, and prognostic factors were evaluated using Cox regression. Results: KRAS mutations were detected in 28.0% (389/1389) of patients. KRAS mutation prevalence was higher in females (32.8% vs 25.8%; P = 0.008), CEA-positive (29.8% vs 24.2%; P = 0.030), CA19-9-positive (31.0% vs 25.6%; P = 0.030), and patients with multiple liver metastases (31.4% vs 23.9%; P = 0.002; Table 1). KRAS mutations were more frequent in right-sided colon primaries (34.9%) compared to left hemicolon (24.5%). Among KRAS-mutant patients (n = 389), codon 12 variants accounted for 62.7%, predominantly G12D (32.1%) and G12V (12.9%) (Supplementary Table 1). Univariate Cox analysis showed that KRAS mutations were associated with shorter PFS (HR = 1.48; P < 0.001) and OS (HR = 2.18; P < 0.001), with the largest hazard ratios for G12V (PFS: HR = 2.06; OS: HR = 3.61; both P < 0.001; Table 2). KRAS-mutant tumors had shorter mPFS and mOS compared to WT tumors (mOS: 43 months vs not reached; mPFS: 15.87 vs 23.93 months; both P < 0.001) (Figure 1A). Codon 12 mutations, particularly G12V, conferred the worst survival outcomes (Figure 1B and 1C). Baseline characteristics were similar between codon 12 and non-codon 12 KRAS-mutant groups, except codon 12 variants were more common in males (66.0% vs 55.3%; P = 0.049; Table 3). In the codon 12–mutant subgroup (n = 244), G12V tumors were more frequent in CEA-positive than CEA-negative patients (25.4% vs 9.3%; P = 0.003) and in those with multiple liver metastases (25.2% vs 13.9%; P = 0.036; Table 4). Conclusions: KRAS mutations were frequent and independently predicted worse OS and PFS after curative hepatectomy. Prognostic impact varied across KRAS subtypes, with G12V indicating a high-risk subgroup. Incorporating codon-specific KRAS information into risk stratification and perioperative decision-making may refine patient selection and trial design.

Abstract Immunotherapy has emerged as a transformative approach to cancer treatment, yet its clinical efficacy in most solid tumours remains limited, largely because of the immunosuppressive tumour microenvironment (TME). In this context, glucose metabolic reprogramming has emerged as a central determinant of tumour progression and immune dysfunction because it not only sustains the proliferative and biosynthetic demands of malignant cells but also profoundly reshapes immune responses within the TME. Effective antitumour immunity depends on the metabolic adaptability of effector immune cells, particularly the coordinated use of glycolysis and oxidative phosphorylation to support activation, expansion and cytotoxic function. Under the nutrient‐deprived, hypoxic and acidic conditions that characterize the TME, however, these cells undergo metabolic restriction that progressively drives dysfunction and exhaustion. By contrast, regulatory T cells, tumour‐associated macrophages and myeloid‐derived suppressor cells exhibit greater metabolic plasticity, enabling their persistence and reinforcing their immunosuppressive activity. In this review, we discuss how glucose metabolic reprogramming drives immune dysfunction through several interconnected processes, including glucose competition, lactate accumulation, reciprocal regulation between glucose metabolism and cytokine signalling, glycosylation remodelling and dynamic crosstalk with immune checkpoint signalling. Collectively, these mechanisms position glucose metabolism as a pivotal immunometabolic axis linking tumour bioenergetics to immune evasion and therapeutic resistance. A deeper understanding of this regulatory network may inform the rational development of combination strategies that integrate metabolic intervention with immunotherapy, ultimately improving therapeutic precision and the durability of clinical benefit. Highlight Glucose metabolic reprogramming is a central driver of immunosuppression in the tumour microenvironment. Glucose competition establishes a selective bioenergetic hierarchy that constrains antitumour immunity. Lactate accumulation and reciprocal regulation with cytokine signalling amplify immunosuppressive signalling and reinforce immune exclusion. Glycosylation remodelling translates altered metabolic flux into sustained changes in receptor stability, ligand recognition and checkpoint responsiveness. Dynamic crosstalk with immune checkpoint signalling entrenches chronic immune dysfunction and therapeutic resistance.

BACKGROUND:Associating liver partition and portal vein ligation for staged hepatectomy (ALPPS) effectively induces rapid liver hypertrophy in patients with initially unresectable liver tumours, yet the immunological mechanisms remain unclear. OBJECTIVE:We aim to elucidate the immune alterations and underlying mechanisms driving liver regeneration following ALPPS. DESIGN:The cohort study included single-cell RNA sequencing (scRNA-seq) and spatial transcriptomics on remnant liver tissues from ALPPS patients. Neutrophil infiltration was validated by flow cytometry and histological analyses in the world's largest ALPPS clinical cohort and mouse ALPPS models. Functional validation, including neutrophil depletion, matrix metalloproteinase 9 (MMP9) inhibition and CD177 blockade, as well as Cd177 knockout and CD177+ neutrophil infusion in vivo. RESULTS:scRNA-seq revealed substantial neutrophil infiltration following stage 1 ALPPS. Depletion of neutrophils impaired liver regeneration. Among subsets, CD177+ neutrophils were metabolically active with enhanced neutrophil extracellular traps formation and secreted MMP9. MMP9 inhibition disrupted extracellular matrix (ECM) degradation and hepatocyte growth factor alpha (HGF-α) release, impairing regeneration. CD177+ neutrophils interacted with endothelial cells via CD177-PECAM1 to facilitate transmigration, while hepatic stellate cell-derived CXCL8 promoted neutrophil chemotaxis via CXCL8-CXCR1/2. Cd177 deficiency attenuated neutrophil infiltration and regenerative growth, while CD177+ neutrophil infusion restored regeneration, which was abolished in Cd177-/- mice. CONCLUSIONS:CD177+ neutrophils drive liver regeneration by promoting endothelial transmigration, ECM degradation and HGF-α release. These findings reveal a neutrophil-mediated mechanism driving surgical liver regeneration and support the potential of CD177+ neutrophil infusion to establish a proregenerative hepatic environment for therapeutic strategies in liver failure.