Multi-omics spatial characteristics of CD8+TRM cells in hepatocellular carcinoma and immunotherapy response prediction

Shi-Peng Li^1*Ju Ma¹Xinqiang Li²Bingbing Qiao³Liancai Wang¹Deyu Li¹Hua Zhou⁴Hua Zhou^5*Jinzhen Cai^6*

• ¹Henan Provincial People's Hospital, Zhengzhou, China
• ²The Affiliated Hospital of Qingdao University, Qingdao, China
• ³The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China
• ⁴The First Affiliated Hospital of Henan University of Science and Technology, Luoyang, China
• ⁵Tianjin Medical University School of Basic Medical Sciences, Tianjin, China
• ⁶Fujian Medical University Union Hospital, Fuzhou, China

Immune checkpoint blockade (ICB), a therapeutic strategy that reinvigorates anti-tumor immune responses by disrupting inhibitory signaling pathways in immune cells, has emerged as a pivotal systemic modality for patients with advanced hepatocellular carcinoma (HCC). However, the fact that only a subset of patients achieve durable clinical responses to ICB underscores the urgent need to develop reliable, clinically actionable predictive biomarkers. Despite the potential of tumor-infiltrating immune cells (TIICs) to stratify this patient cohort, the predictive utility of TIICs with respect to their density and spatial arrangement has not been conclusively validated for clinical application. CD8+T cells frequently infiltrate the HCC microenvironment and play a critical role in immunotherapy, we developed and validated a model based on contrast enhanced computed tomography (CECT) images and preliminarily predicted the change of CD8+T cells distribution of HCC patients after ICB treatment, then through single-cell RNA sequencing analysis of HCC tumor tissues, we clearly show that CD8+ T cells and CD8+ tissue-resident memory T cells (TRM) undergo a progressive and statistically significant upregulation in their proportions post-ICB treatment. What's more, multiplex immunohistochemistry (mIHC) was used to develop a multidimensional TIIC signature, which included quantifying the proportions of CD8+TRM, CD4+TRM, and CD68+ cells, as well as analyzing the spatial organization of CD8+PD-1+TRM cells. We found that the proportion and infiltration of CD8+TRM cells in HCC tissue increased from the immune margin (IM) to the tumor core (TC) after treatment with anti-PD-1/PD-L1 immunotherapy. These results indicate that the spatial heterogeneity of CD8+TRM cells within the tumor immune microenvironment critically determines immunotherapy efficacy in HCC.