FIGURE 2 from Characterizing Neutrophil Subtypes in Cancer Using scRNA Sequencing Demonstrates the Importance of IL1β/CXCR2 Axis in Generation of Metastasis-specific Neutrophils

Characterization of neutrophils in metastasis. A, UMAP of neutrophils in CRCLM. B, Coexpression of Hₑnriched and Tₑnriched signatures. One cluster is not enriched for either signature (blue arrow). C and D, Unsupervised pseudotime analysis and estimated smoothers for TXNIP expression over the different numbered pseudotime lineages. E, Coexpression of TXNIP and CXCR2. F and G, Expression and estimated smoothers for CXCL8 over pseudotime. H, Coexpression of CXCL8 and IL1β. I, IHC staining of TXNIP in a patient CRCLM sample at 4x (left) and 10x (right). Scale bars = 50 µm. J and K, IHC staining of ITGAM (Neutrophils) and CD3 (T cells) in a patient CRCLM sample at 4x, scale bars = 50 µm. Dashed squares indicate regions where immune cells cluster. L, Differentially expressed genes in metastasis-specific neutrophil cluster. M, Hₑnriched, Tₑnriched, and Mₑnriched gene signatures in mouse bulk-RNA-seq neutrophil dataset. L, Healthy liver tissue, PT: Primary tumor, LMET: Liver metastasis. N, Averaged expression of the individual genes of the three signatures. O, UMAP plot of integrated human neutrophils from PT and metastatic (M) datasets of different cancers. P, Coexpression of CXCR2 with IL1β (top) and TXNIP (bottom). Q, Differential gene expression between neutrophils in malignancy compared with PT. R and S, GO and KEGG analysis of MCRC neutrophils.