Spatial proximity of Tumor-Immune interactions predicts patient outcome in hepatocellular carcinoma

The fitness and viability of a tumor ecosystem are influenced by the spatial organization of its cells. We aimed to study the structure, architecture, and cell-cell dynamics of the heterogenous liver cancer tumor microenvironment using spatially resolved multiplexed imaging.We performed co-detection by indexing (CODEX) multiplexed immunofluorescence imaging on 68 hepatocellular carcinoma (HCC) biopsies from Thai patients (TIGER-LC) as a discovery cohort, and then validated the results in an additional 190 HCC biopsies from Chinese patients (LCI). We segmented and annotated 117,270 and 465,632 cells from the TIGER-LC and LCI cohorts, respectively. We observed four groups of TIGER-LC patients (IC1, IC2, IC3, IC4) with distinct tumor-immune cellular interaction patterns. In addition, patients from IC2 and IC4 had much better overall survival than those from IC1 and IC3. Noticeably, tumor and CD8+ T cell interactions were strongly enriched in IC2, the group with the best patient outcomes. Close proximity between tumor and CD8+ T cells was a strong predictor of patient outcome in both the TIGER-LC and the LCI cohorts. Bulk transcriptomic data from 51 of the 68 HCC cases was utilized to determine tumor-specific gene expression features of our classified subtypes. Moreover, we observed that the presence of immune spatial neighborhoods in HCC as a measure of overall immune infiltration is linked to better patient prognosis.Highly multiplexed imaging analysis of liver cancer reveals tumor-immune cellular heterogeneity within spatial contexts, such as tumor and CD8+ T cell interactions, which may predict patient survival.