Cell-Autonomous Cxcl1 Sustains Tolerogenic Circuitries and Stromal Inflammation via TNF in Pancreatic Cancer

We have shown that KRAS-TP53 genomic coalteration is associated with immune -excluded microenvironments, chemoresistance, and poor survival in pancreatic ductal adenocarcinoma (PDAC) patients. By treating KRAS-TP53 cooperativity as a model for high -risk biology, we now identify cell-autonomous Cxcl1 as a key mediator of spatial T-cell restriction via interactions with CXCR2+ neutrophilic myeloid-derived suppressor cells in human PDAC using imaging mass cytometry. Silencing of cell-intrinsic Cxcl1 in LSL-KrasG12D/+;Trp53R172H/+;Pdx-1Cre/+(KPC) cells reprograms the traffi cking and functional dynamics of neutrophils to overcome T-cell exclusion and controls tumor growth in a T cell-dependent manner. Mechanistically, neutrophil-derived TNF is a cen-tral regulator of this immunologic rewiring, instigating feed-forward Cxcl1 overproduction from tumor cells and cancer-associated fi broblasts (CAF), T-cell dysfunction, and infl ammatory CAF polarization via transmembrane TNF-TNFR2 interactions. TNFR2 inhibition disrupts this circuitry and improves sensitivity to chemotherapy in vivo . Our results uncover cancer cell-neutrophil cross-talk in which context-dependent TNF signaling amplifi es stromal infl ammation and immune tolerance to promote therapeutic resistance in PDAC.SIGNIFICANCE: By decoding connections between high-risk tumor genotypes, cell-autonomous infl am-matory programs, and myeloid-enriched/T cell-excluded contexts, we identify a novel role for neu-trophil-derived TNF in sustaining immunosuppression and stromal infl ammation in pancreatic tumor microenvironments. This work offers a conceptual framework by which targeting context-dependent TNF signaling may overcome hallmarks of chemoresistance in pancreatic cancer.