Background and Aims: HCC remains a leading cause of cancer mortality in China. Anti-angiogenic drugs (AADs) plus immune checkpoint blockade (ICB) combination therapy shows considerable promise for HCC; however, its efficacy is hampered by immunosuppression within the tumor microenvironment. High endothelial venules (HEVs) facilitate lymphocyte migration and tumor infiltration. The aim was to study the formation, functional mechanisms, and clinical relevance of HEVs in the treatment of HCC with combination therapy. Approach and Results: Single-nucleus RNA sequencing, flow cytometry, and immunohistochemistry revealed increased HEV expression and higher CD3 ⁺ T-cell infiltration in HCC tissue after a combination of AAD and ICB therapy. Multiplex immunohistochemistry and spatial analysis demonstrated that CCR7 ⁺ CD8 ⁺ T cells were spatially associated with HEVs. Pseudotime analysis of human T cells and treatment of Hepa1-6 orthotopic liver tumor mouse models with CCR7 ⁺ CD8 ⁺ T-cell transfusions were used to show that CCR7 ⁺ CD8 ⁺ T cells can differentiate into cytotoxic effector T cells. The same models demonstrated that combination therapy activated VEGFC and non-canonical NF-κB pathways, promoting HEV formation. Kaplan–Meier analysis revealed that high HEV density correlated with improved clinical response and prolonged survival. Conclusions: HEVs are pivotal in modulating immune activity within the HCC tumor microenvironment. Targeting the VEGFC–NF-κB (non-canonical)–HEV axis could be a promising therapeutic strategy to enhance antitumor immunity and improve outcomes in patients with HCC who are receiving combination AAD plus ICB therapy.

Large hepatocellular carcinoma (HCC) exhibits heterogeneous morphologies and varied responses to treatment. We evaluated outcomes of patients with different large HCC classifications receiving surgical resection (SR) or transarterial chemoembolization plus ablation (TA). Patients with HCC ≥ 5 cm receiving SR or TA between May 2016 and December 2020 at one center were analyzed retrospectively and with propensity score matching (PSM). Overall survival (OS) and progression-free survival (PFS) of the 2 treatment groups were compared. Tumors were classified according to imaging morphology and gross pathology: Type I, simple nodular; Type II, simple nodular with extranodular growth or confluent multinodular; Type III, infiltrative. Of 644 patients, 374 met the inclusion criteria (300 received SR and 74 received TA). Before PSM, median follow-up was 51.2 (IQR 29.6-65.3) months, and the SR group had longer OS (HR 2.13, 95% CI 1.44-3.15, p<0.001) and PFS (HR 2.31, 95% CI 1.66-3.20, p0.05). Infiltrative HCC (Type III) was an independent negative prognostic factor for OS and PFS. Within both treatment groups, patients with infiltrative HCC had shorter OS and PFS than patients with non-infiltrative HCC (Types I and II) (all p<0.001). For patients with HCC ≥ 5 cm, tumor classification is an important prognostic factor. In patients with non-infiltrative HCC, TA and SR had comparable OS after PSM. For patients with infiltrative HCC, TA and SR had limited efficacy.

The Coatomer protein complex Zeta 1 (COPZ1) has been reported to play an essential role in maintaining the survival of some types of tumors. In this study, we sought to explore the molecular characteristics of COPZ1 and its clinical prognostic value through a pan-cancers bioinformatic analysis. We found that COPZ1 was extremely prevalent in a variety of cancer types, and high expression of COPZ1 was linked to poor overall survival in many cancers, while low expression in LAML and PADC was correlated with tumorigenesis. Besides, the CRISPR Achilles’ knockout analysis revealed that COPZ1 was vital for many tumor cells’ survival. We further demonstrated that the high expression level of COPZ1 in tumors was regulated in multi-aspects, including abnormal CNV, DNA-methylation, transcription factor and microRNAs. As for the functional exploration of COPZ1, we found a positive relationship between COPZ1’s expression and stemness and hypoxia signature, especially the contribution of COPZ1 on EMT ability in SARC. GSEA analysis revealed that COPZ1 was associated with many immune response pathways. Further investigation demonstrated that COPZ expression was negatively correlated with immune score and stromal score, and low expression of COPZ1 has been associated to more antitumor immune cell infiltration and pro-inflammatory cytokines. The further analysis of COPZ1 expression and anti-inflammatory M2 cells showed a consistent result. Finally, we verified the expression of COPZ1 in HCC cells, and proved its ability of sustaining tumor growth and invasion with biological experiments. Our study provides a multi-dimensional pan-cancer analysis of COPZ and demonstrates that COPZ1 can serve as both a prospective target for the treatment of cancer and a prognostic marker for a variety of cancer types.