Funding
National Key Research and Development Program of China (NKPs)
National Natural Science Foundation of China (NSFC)
National Natural Science Foundation of China-Guangdong Joint Fund (国家自然科学基金-广东省联合基金)
Guangdong Basic and Applied Basic Research Foundation
Open Fund Project of Guangdong Academy of Medical Sciences
Key-Area Research and Development Program of Guangdong Province
Science and Technology Planning Project of Guangdong Province (S&T Project of Guangdong Province)
Science and Technology Projects in Guangzhou
Fundamental Research Funds for the Central Universities (Fundamental Research Fund for the Central Universities)
ARTICLE ABSTRACT
Despite the pivotal role of CTLs in antitumor immunity, a substantial proportion of CTL-rich patients with hepatocellular carcinoma (HCC) experience early relapse or immunotherapy resistance. However, spatial immune variations impacting the heterogeneous clinical outcomes of CTL-rich HCCs remain poorly understood. In this study, we compared the single-cell and spatial landscapes of 20 CTL-rich HCCs with distinct prognoses using multiplexed in situ staining and validated the prognostic value of myeloid spatial patterns in a cohort of 386 patients. Random forest and Cox regression models identified macrophage aggregation as a distinctive spatial pattern characterizing a subset of CTL-rich HCCs with an immunosuppressive microenvironment and poor prognosis. Integrated analysis of single-cell and spatial transcriptomics, combined with in situ staining validation, revealed that spatial aggregation enhanced protumoral macrophage reprogramming in HCCs, marked by lipid metabolism orientation, M2-like polarization, and increased adjacent CTL exhaustion. This spatial effect on macrophage reprogramming was replicated in HCC-conditioned human macrophage cultures, which showed an enhanced capability to suppress CTLs. Notably, increased macrophage aggregation was associated with higher response rates to anti–PD-1 immunotherapy. These findings suggest that the spatial distribution of macrophages is a biomarker of their functional diversities and microenvironment status, which holds prognostic and therapeutic implications.
