Yanchun Wang, Guangyu Zhang, Xin Zhang, Gaoke Liu, Lihua Zhang, Lei Chen, Shuli Sang, Sheng Yao, Yang Fei, Zhichao Tian, Yu Zang, Hongwei Wang, Yiping Dong, Chunjie Liu, Xizhao Chen, Yan Liu, Min Zhang
Tertiary lymphoid structures (TLSs) have emerged as critical prognostic and immunotherapeutic indicators in cancer, with their clinical significance modulated by spatial distribution patterns and density. Here, we performed integrated single-cell and spatial transcriptomic profiling of 30 gastric cancer (GC) specimens stratified by TLS spatial localization patterns. Comparative analysis shows pronounced enrichment of CXCL13+ T lymphocytes (TLCs), CXCR5+ germinal center B lymphocytes (gc_B cells), LAMP3+CD80+ activated dendritic cells (DCs), and SELP+ACKR1+ high endothelial venule (HEV) cells within intratumoral-TLS (iTLS) rich tumors compared to peritumoral-TLS (pTLS) and desert-TLS (dTLS) tumor subtypes. Multimodal cell–cell interaction analysis and functional experiments demonstrate that HEV expressed VCAM1 and ICAM1 recruits and activates CXCL13+ TLC through the CXCL13-ACKR1 pathway, which promotes TLS formation via CXCL13-CXCR5-dependent crosstalk with B lymphocytes. We further develop a single-cell/spatial TLS signature that captures the cellular ecosystem of iTLS-containing tumor, demonstrating predictive value for immunotherapy outcomes in GC patients. Tertiary lymphoid structures (TLS) within the tumour microenvironment are important for prognostic and immunotherapeutic indications in cancer. Here the authors use a spatial and single cell transcriptomics analysis of gastric cancer (GC) samples and show transcriptomic differences grouped by types of TLS and generate a prognostic transcriptomic signature to help predict outcome in GC patients.