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.

PURPOSE:Neurofibromatosis type 2 (NF2) is a tumor predisposition syndrome characterized by bilateral vestibular schwannomas (VSs) resulting in deafness and brainstem compression. This study evaluated efficacy and biomarkers of bevacizumab activity for NF2-associated progressive and symptomatic VSs.PATIENTS AND METHODS:Bevacizumab 7.5 mg/kg was administered every 3 weeks for 46 weeks, followed by 24 weeks of surveillance after treatment with the drug. The primary end point was hearing response defined by word recognition score (WRS). Secondary end points included toxicity, tolerability, imaging response using volumetric magnetic resonance imaging analysis, durability of response, and imaging and blood biomarkers.RESULTS:Fourteen patients (estimated to yield > 90% power to detect an alternative response rate of 50% at alpha level of 0.05) with NF2, with a median age of 30 years (range, 14 to 79 years) and progressive hearing loss in the target ear (median baseline WRS, 60%; range 13% to 82%), were enrolled. The primary end point, confirmed hearing response (improvement maintained ≥ 3 months), occurred in five (36%) of 14 patients (95% CI, 13% to 65%; P < .001). Eight (57%) of 14 patients had transient hearing improvement above the 95% CI for WRS. No patients experienced hearing decline. Radiographic response was seen in six (43%) of 14 target VSs. Three grade 3 adverse events, hypertension (n = 2) and immune-mediated thrombocytopenic purpura (n = 1), were possibly related to bevacizumab. Bevacizumab treatment was associated with decreased free vascular endothelial growth factor (not bound to bevacizumab) and increased placental growth factor in plasma. Hearing responses were inversely associated with baseline plasma hepatocyte growth factor (P = .019). Imaging responses were associated with high baseline tumor vessel permeability and elevated blood levels of vascular endothelial growth factor D and stromal cell-derived factor 1α (P = .037 and .025, respectively).CONCLUSION:Bevacizumab treatment resulted in durable hearing response in 36% of patients with NF2 and confirmed progressive VS-associated hearing loss. Imaging and plasma biomarkers showed promising associations with response that should be validated in larger studies.

Supplementary Fig. S9: Expression levels of Cxcr3 and its ligands are increased in ICC tissues after GC/dual ICB treatment in murine 425-ICC.

De novo vessel formation (vasculogenesis) in vitro is a key step in tissue engineering to preserve tissue viability for long-term assays and testing therapeutic agents. However, in vitro vasculogenesis is often unreliable due to differences in vascular-supporting cells, including endothelial cells and stromal cells such as smooth muscle cells (SMCs) and fibroblasts. Here, we developed a robust co-culture system of HUVECs and SMCs to generate stable vascular networks capable of maintaining tissue viability over extended periods. Given that SMC plasticity is a major limitation in supporting endothelial network formation, we systematically evaluated the effects of passage number, confluency, and freezing on primary SMC function. To overcome this limitation, we generated immortalized supportive SMCs, which preserved their vasculogenic gene program and functional capacity even at high passage. In addition, we identified and validated key genes associated with endothelial support, including CD248, C3, and FBLN1, all essential for vasculogenesis. Immortalized SMCs consistently maintained expression of these genes and supported robust vessel formation under variable culture conditions. Collectively, this study demonstrates that immortalized SMCs provide a stable, reproducible platform for endothelial-SMC co-cultures, enabling long-term vascularized tumor models suitable for functional studies and therapeutic screening.

Supplementary Fig. S8: Bulk tissue RNA sequencing analysis of ICC after GC/dual ICB combination therapy in orthotopic murine 425-ICC model.

Supplementary Fig. S12: Effect of ICB treatment scheduling on efficacy and toxicity.

Abstract Background/Aim: Immune checkpoint inhibitors (ICs) have improved survival outcomes in advanced hepatocellular carcinoma (HCC), but most HCC patients experience treatment resistance. Thus, we aimed to stratify HCC patients to target therapy-resistant TICs and identify responders through stemness mRNA profiling. Our second aim was to evaluate novel CTC signatures to predict treatment efficacy in response to standard anti-VEGFA+anti-PD-L1 antibody immunotherapy with cancer-driver-targeted chemotherapies. Methods: The Genome-wide CRISPR-Cas9 knockout (GeCKO) method was used to identify genes required for killing tumors expressing TP53, CTNNB1, and ARID2. To confirm the drug efficacy, we used CRISPR/Cas9-mediated driver gene knock-ins and knockouts combined with alcohol Western diet feeding to promote HCC development in a humanized FNRG (Fah- / -;Nod;Rag1- / -;Il2rgc- / - ) mouse harboring ARID1A mutation or TP53 mutation in a PDX Model. Drugs paired with ICI were tested for treatment of HCC in genetically defined PDX and these humanized FNRG mice. Results: The GeCKO screening in ARID1A-mutant HCC showed that a combination of a Polycomb repressive complex 2 (PRC2) inhibitor, anti-PD-L1, and anti-VEGFA treatment reduced tumor size and extended survival in ARID1A-mutant PDX mice with humanized immune systems. The most successful combination from these approaches, comprising anti-EZH1/2 inhibitor + anti-PD-L1 + anti-VEGFA for ARID1A mutant HCC or all-trans retinoic acid (ATRA) and an HDAC inhibitor, along with anti-PD-L1, for TP53 mutant HCC-eradicated human patient-derived liver TICs grown ex vivo from tumors generated in mice. Single-cell RNA-seq analyses of patient HCCs identified 16 upregulated gene signatures with statistical significance. Principal component analyses provide a clear separation between Barcelona Clinic Liver Cancer (BCLC) stage categories A (benign) and B (malignant). Alcohol Western diet-associated TLR4-AKT signal leads to EZH2 phosphorylation of PRC2 complex to switch from histone suppressive activity to transactivation. The most effective targeting of PRC2 components, which eliminated human patient-derived liver TICs grown ex vivo in mice. Conclusions: Our innovative approach, using precision medicine to identify patients most likely to respond to specific drug combinations targeting TICs and cancer-driver mutations, will revolutionize cancer treatment by addressing a core component of therapeutic resistance in HCC. Citation Format: Yang Cui, Alexandria Borges, Meng Li, Linda Sher, Bangyan Stiles, Dan G. Duda, Anthony El-Khoueiry, Keigo Machida. CTC/cfDNA biomarker-guided precision medicine to overcome immune checkpoint inhibitor resistance in hepatocellular carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(8_Suppl):Abstract nr LB182.

Supplementary Table S1: Antibody list.

Cancer immunotherapy relies on PD-L1 expression, often assessed through immunohistochemistry, offering limited insights. In-vivo fluorescence lifetime (FLT) imaging, utilizing PD-L1-targeted probes, provides quantitative estimates of PD-L1 heterogeneity across tumors, showing promise for clinical translation of FLT imaging.

Supplementary Fig. S2: Standard chemotherapy converts ICB-resistant ICCs to ICB-responsive tumors, significantly delays tumor progression and increases survival in mice.

Supplementary Fig. S10: Schematic of the animal experiment design.

Supplementary Fig. S6: CTLA-4 blockade mediates the efficacy of GC/ICB therapy in ICC and increases CD8+CTL frequency in murine ICC.

This file contains all the supplementary figures and tables related to this manuscript.

Hepatocellular carcinoma (HCC) represents one of the leading contributors to cancer-related deaths, with the majority of patients diagnosed at stages where curative treatment is no longer possible. Combining stereotactic body radiotherapy (SBRT) with immune checkpoint inhibition (ICI) has gained increasing attention as a therapeutic approach. Beyond its ability to provide high local tumor control (LC), SBRT can provoke immunogenic tumor cell death, promote antigen release and presentation, and modulate the tumor microenvironment in ways that enhance systemic antitumor immunity.In this narrative review, we outline the scientific rationale for integrating SBRT with ICIs, discuss mechanistic and translational findings and summarize results from key clinical trials. The currently available data indicate a synergistic interaction, most notably reflected in improved survival and response rates. Nevertheless, variability in dose and fractionation schedules, treatment sequencing, and patient characteristics complicates interpretation. Well-designed prospective studies are needed to establish optimal protocols and identify predictive biomarkers to guide patient selection.

Abstract Group 3 medulloblastomas (G3MB) carry the worst prognosis among medulloblastoma subtypes, yet molecularly targeted therapies remain elusive. Standard treatments cause severe long-term morbidity in survivors. Here, we identify tumor-derived sphingosine kinase 2 (SPHK2) as an essential driver of G3MB initiation and progression. SPHK2 exacerbates local immunosuppression by suppressing cytotoxic T-cell and NK-cell activity while promoting regulatory T-cell infiltration. Genetic or pharmacologic SPHK2 inhibition using Opaganib attenuates pro-survival tumor signaling and restores anti-tumor immunity, significantly improving survival in syngeneic G3MB mouse models. Combining Opaganib with fractionated low-dose radiation (f-LDRT) further enhances antigen presentation and reprograms tumor-associated myeloid cells toward an anti-tumor phenotype. This combination therapy markedly prolongs survival without inducing significant toxicity. Overall, our study establishes SPHK2 as a previously unrecognized therapeutic target and presents a safe, effective, microenvironment-reprogramming regimen for G3MB. One Sentence Summary Direct inhibition of tumor-derived SPHK2 overcomes local immunosuppression and downregulates pro-survival signaling in Group 3 medulloblastoma, while combination with fractionated low-dose radiation further enhances anti-tumor immunity and significantly improves survival.

Supplementary Fig. S11: Cxcr3 in CD8 T cells mediates the benefit of GC/ICB combination therapy in orthotopic murine 425-ICC model.

PURPOSE:To investigate the impact of radiation therapy (RT) for brain tumors on circulating lymphocyte populations and understand this from a dosimetric perspective. METHODS AND MATERIALS:This prospective study enrolled patients with primary brain tumors treated with either photon or proton RT. To specifically study the effect of RT, only patients without concurrent chemotherapy were eligible. For each patient, blood samples were collected before, during, and after RT, as well as at the first follow-up, for flow cytometry analysis of blood cells. We used a stochastic model to compute blood dose distributions from treatment plans and patient-specific blood flow simulations. Dose to the lymph nodes (LNs) in the head and neck region was also assessed. Dose distributions and lymphocyte trends from patients treated with photon versus proton therapy were compared. RESULTS:Thirty-five patients with a median age of 50 were included: 27 patients with benign tumors (mostly meningiomas) and 8 with malignant tumors (mostly gliomas). There were no significant RT-induced changes in either the total lymphocyte count or its subpopulations in this patient cohort. In line with this finding, dosimetric analyses showed that total blood doses were ∼0.1 to 0.2 Gy and that most LNs in the head and neck area are spared. Proton RT was associated with a greater dosimetric sparing of both blood and LNs. CONCLUSIONS:In this exploratory analysis, we found no evidence that RT alone results in significant changes in peripheral lymphocyte counts in patients with primary brain tumors treated to limited intracranial volumes. The lack of apparent RT-induced cytotoxicity is consistent with the limited dose received by either the blood or the regional LNs.