Spatial multi-omics reveals that CXCL10 enhances the response to nICT in surgically resectable esophageal squamous cell carcinoma

Abstract Neoadjuvant immune-chemotherapy (nICT) has transformed the treatment strategy for esophageal squamous cell carcinoma (ESCC), yet the molecular mechanisms underlying treatment response remain incompletely understood. To investigate these mechanisms, we jointly analyzed matched ESCC specimens obtained before and after neoadjuvant therapy using single-cell transcriptomic profiling, spatial transcriptomics, and spatial metabolomic imaging. This multilayered, same-sample framework revealed extensive remodeling of the tumor microenvironment, including enhanced immune activation and marked changes in the spatial organization of major cellular populations. Notably, CXCL10 increased the intratumoral proportion of CD8⁺ T cells, promoted their cytotoxic activation, and enhanced T-cell–mediated tumor elimination both in vitro and in vivo. Genetic silencing or pharmacological inhibition of CXCR3 attenuated these effects and diminished the therapeutic benefit of anti-PD1 treatment, supporting a functional role for the CXCL10–CXCR3 axis in modulating immunotherapy efficacy. Spatial metabolomics further revealed pronounced remodeling of ornithine metabolism in malignant regions, characterized by suppression of the urea cycle, potential involvement of a glutamate/proline-linked ornithine pathway, and enhanced polyamine synthesis. An ornithine metabolism-related gene signature was strongly associated with poor prognosis in the TCGA-ESCC cohort, while pharmacological inhibition of ornithine decarboxylase enhanced the antitumor efficacy of anti-PD1 therapy in vivo. Together, these findings provide a spatial multi-omics atlas of ESCC following nICT and identify the CXCL10–CXCR3 axis and ornithine-centered metabolic remodeling as potentially actionable immune and metabolic features associated with treatment response.