Cancer-associated Fibroblasts in Pan-Cancer Drive CXCR2+VNN2+ Neutrophils Reprogramming to Mediate Immunosuppression and Immunotherapy Resistance

Neutrophils are the most abundant granulocyte population and have important functions such as defense against pathogens. However, they show significant heterogeneity and play more complex roles in tumors. The theory of two-tiered differentiation of neutrophils is insufficient to summarize their phenotypic and functional heterogeneity. Therefore, specific regulatory mechanisms remain to be explored and neutrophil-based therapeutic regimens remain challenging. Here, we generated a single-cell atlas of neutrophils from 462 patients with 21 cancer types, revealing their heterogeneity, with CXCR2+ VNN2+ Neu as the main functional subpopulation exerting immunosuppressive effects. Spatial transcriptomic data from the pan-cancer elucidated that fibroblast regulated the phenotypic shift of CXCR2+ VNN2+ Neu in tumor tissues and enabled it to acquire immunosuppressive functions through receptor ligands, cytokines, and extracellular vesicles, which suggested that the tumor microenvironment component was a key reason for the heterogeneity of the prognostic association between neutrophils and pan-cancer patients. Subsequently, we constructed a gene regulatory network to demonstrate the specific regulatory mechanisms of this subpopulation and confirmed that the relevant transcription factors were closely associated with its immunosuppressive function. The pan-cancer immunotherapy cohort proved that the CXCR2+ VNN2+ Neu phenotypic shift was also an important cause of immunotherapy resistance in patients. We finally constructed a deep learning model named Deepsurv to accurately stratify pan-cancer patients based on the CXCR2+ VNN2+ Neu phenotypic shift gene regulatory network (CVN-GRN) and predict the prognosis of the patients, which achieved the desired results.