Yaofei Jiang

Elucidating how Epstein–Barr virus (EBV) shapes tumor–immune interactions is important for improving therapy in nasopharyngeal carcinoma (NPC). This multi‐cohort analysis integrates single‐cell, spatial, and bulk transcriptomic data with multiplex immunofluorescence and functional assays. Single‐cell profiling identifies stemness‐associated epithelial states and enrichment of SPP1 ⁺ tumor‐associated macrophages (TAMs) in EBV ⁺ NPC. SPP1 ⁺ TAM abundance is associated with stemness‐associated epithelial features and dysfunctional T‐cell states, while high MIF and SPP1 ⁺ macrophage signatures are associated with shorter progression‐free survival. Within the EBV ⁺ spatial cohort, SPP1 ⁺ TAMs co‐occur with stemness‐associated tumor cells, exhausted CD8 ⁺ T cells, and regulatory T cells in Niche 5, which is more frequent in non‐response tumors following chemoimmunotherapy. Functional experiments show increased MIF expression and secretion by EBV ⁺ epithelial cells. MIF inhibition attenuates conditioned‐medium‐induced monocyte migration and SPP1‐associated TAM‐like programming, whereas recombinant MIF reproduces these effects. Conversely, macrophage‐derived SPP1 increases stemness‐associated marker expression and tumorsphere formation in EBV ⁺ epithelial and tumor models. It also impairs primary CD8 ⁺ T‐cell proliferation, effector function, and cytotoxic activity while increasing exhaustion‐associated markers; CD44 blockade partially reverses these effects. Collectively, these findings support an EBV‐associated MIF–SPP1 macrophage–tumor/T‐cell crosstalk model linking stemness‐associated epithelial phenotypes with immune dysfunction and adverse clinical outcomes.

Background: Combinatory therapeutic strategy containing immunochemotherapy as part of induction therapy components is one of the current trends in the treatment of high‐risk metastatic locally advanced nasopharyngeal carcinoma (NPC). However, the mechanism underlying the heterogeneity of response at the single‐cell level has not been underexplored.Methods: 18 bulks and 11 single‐cell RNA sequencing from paired before‐treatment and on‐treatment samples in patients with treatment‐naive high‐risk metastatic locally advanced NPCs were obtained. Following quality control, a total of 87 191 cells were included in the subsequence bioinformatics analysis.Results: Immunochemotherapy was associated with on‐treatment tumour microenvironment (TME) remodelling, including upregulation of anti‐TMEs signatures, downregulation of pro‐TMEs signatures, reversing CD8⁺ T exhaustion, and repolarizing proinflammatory TAMs. For the patients achieving a complete response, the cytotoxic activity of CD8⁺ T cells was stimulated and more interferon‐gamma was provided, which would be the key for TAMs proinflammatory repolarization and eventually promote the CD8⁺ T cells maturation in turn. Among patients who did not reach complete response, differentiation and hypoxia signatures for endothelial cells were elevated after therapy. These patients exhibited higher levels of immune checkpoint genes in malignant cells at the baseline (before treatment), and decreased tumour antigen presentation activity, which may underlie the resistance mechanism to therapy.Conclusions: This study pictures a map of TME modulation following immunochemotherapy‐based combination induction therapy and provides potential future approaches.Highlights: Immunochemotherapy remodeled T cell phenotypes. For the patients achieving complete response, more interferon gamma was provided by CD8⁺ T cells after therapy, which would be the key for TAMs pro‐inflammatory repolarization and eventually promote the CD8⁺ T cells maturation in turns. Among patients who did not reach complete response, malignant cells exhibited higher level of immune checkpoint genes before therapy, and decreased tumor antigen presentation activity, which may underlie the resistance mechanism to therapy.