Stromal spatial signatures of the classical adenosine pathway dictate prognosis in hepatocellular carcinoma

Background The classical adenosinergic pathway (CD39/CD73/A2BR) plays a pivotal role in regulating tumor immune evasion in hepatocellular carcinoma (HCC). However, the prognostic significance of the specific spatial coordination of these enzymes within the tumor microenvironment remains elusive.Methods We retrospectively included 92 patients with HCC who underwent curative hepatectomy. Multiplex immunohistochemistry was used to assess the in situ expression of CD39, CD73, A2BR, and cytokeratin (CK). Digital pathology was employed to segment the tissue into tumor parenchyma (CK+) and stromal regions (CK-). Associations between spatial phenotypes and survival outcomes were evaluated using Kaplan-Meier and Cox proportional hazards models. Furthermore, the transcriptomic profiles of ENTPD1 (CD39), NT5E (CD73), and ADORA2B (A2BR) were analyzed in the TCGA-LIHC cohort (n = 371) to externally validate the protein-level findings.Results Although the densities of CD39, CD73, and A2BR were higher in the tumor parenchyma than those in the stroma, multivariate Cox regression revealed that only stromal CD39⁺CD73⁺A2BR⁺ co-expression phenotype emerged as an independent protective factor for overall survival (hazard ratio [HR] = 0.300, P = 0.035). Conversely, the stromal CD39⁻CD73⁺A2BR⁻ phenotype was identified as an independent risk factor (HR = 2.93, P = 0.023). Correlation analysis further demonstrated a strong positive correlation between CD73 and A2BR within the stroma (r = 0.803, P Conclusion The spatial organization of the adenosine pathway, particularly within the stroma, is associated with HCC prognosis and may provide greater prognostic information than individual marker expression. These findings provide a potential basis for spatially informed risk stratification and may inform future studies of metabolic immunotherapy in HCC.