Tumor-derived JAG2 programs macrophages via NOTCH3 to drive perineural invasion in colorectal cancer

Perineural invasion (PNI) is an important route of dissemination in colorectal cancer (CRC) and is associated with recurrence and poor prognosis, but the underlying immune-tumor mechanisms remain insufficiently defined. We integrated patient-derived data with in vitro and in vivo models to investigate the role of tumor-macrophage signaling in CRC-associated PNI. Single-cell transcriptomic and ligand-receptor analyses identified tumor-derived JAG2 and macrophage NOTCH3 as a PNI-associated communication axis, with enrichment of JAG2-positive tumor epithelial cells and NOTCH3-positive macrophages in nerve-adjacent regions. Functional studies showed that activation of this axis induced STAT3 phosphorylation and increased CCL2 expression in macrophages, supporting STAT3-mediated transcriptional upregulation of CCL2. These macrophages exhibited an immunosuppressive M2-like phenotype with increased neurotrophic and chemotactic mediators and promoted tumor migration and neurotropic behavior in vitro. Disruption of JAG2-NOTCH3 signaling, STAT3 inhibition, or blockade of the CCL2 arm attenuated these effects. Across in vivo models, pathway inhibition reduced tumor growth, decreased CD206-positive tumor-associated macrophages, and attenuated sciatic nerve invasion. Together, these findings identify a targetable JAG2-NOTCH3-STAT3-CCL2 signaling programmed through which CRC cells reprogramed macrophages to establish a neurotropic microenvironment permissive for PNI, suggesting potential therapeutic strategies to limit neural dissemination in CRC.