Maturation, developmental site, and pathology dictate murine neutrophil function

Abstract

Neutrophils have been implicated in poor outcomes in cancer and severe inflammation. We found that neutrophils expressing intermediate levels of Ly6G (Ly6GInt) were present in mouse cancer models and more abundant in those with high rates of spontaneous metastasis. Maturation, age, tissue localization and functional capacity all drive neutrophil heterogeneity. Recent studies have proposed various markers to distinguish between these heterogeneous sub-populations; however, these markers are limited to specific models of inflammation and cancer. Here, we identify and define Ly6G expression level as a robust and reliable marker to distinguish neutrophils at different stages of maturation. Ly6GInt neutrophils were bona fide immature neutrophils with reduced immune regulatory and adhesion capacity. Whereas the bone marrow is a more recognised site of granulopoiesis, the spleen also produces neutrophils in homeostasis and cancer. Strikingly, neutrophils matured faster in the spleen than in the bone marrow with unique transcriptional profiles. We propose that developmental origin is critical in neutrophil identity and postulate that neutrophils that develop in the spleen supplement the bone marrow by providing an intermediate more mature reserve before emergency haematopoiesis.

CXCR2 inhibition enables NASH-HCC immunotherapy

“…7g, h ). Finally, these AZD5069/anti-PD1 TANs closely resembled a recently objectively characterised acute-inflammatory immature-Ly6G Int neutrophil population isolated from lipopolysaccharide-(LPS)-treated mice 50 ( Supplementary Data Fig. 7i ).…”

“…Hence, TAN-enriched immune hubs observed in AZD5069/anti-PD1 treated tumours are able to avoid immune checkpoint inhibition signals that might otherwise cause immune exhaustion. AZD5069/anti-PD1 TANs also displayed a strong correlation with transcriptional changes seen in neutrophils during an acute systemic inflammatory response 50 , including expression of genes involved in exocytosis, myeloid cell activation and degranulation ( Supplementary Data Fig. 7g, h ).…”

Early Neutrophilia Marked by Aerobic Glycolysis Sustains Host Metabolism and Delays Cancer Cachexia

“…We aimed to confirm our findings by analyzing publicly available data on neutrophils, and, therefore, used bulk RNA sequencing data from splenic neutrophils that were isolated from wild-type C57BL/6J and KPC mice (SRA accession number PRJNA749728) [ 16 ]. Unsupervised differential gene expression analysis demonstrated the elevated expression of genes that are associated with hypoxia, apoptosis, fatty acid, and cholesterol metabolism, as well as the overexpression of genes that are related to glycolysis and oxidative phosphorylation in neutrophils of the KPC mice compared to neutrophils of the wild-type control mice without cancer ( Figure S2E ).…”

“…Bulk RNA sequencing data (FASTQ files) corresponding to neutrophils that were isolated from the spleen of wild-type C57BL/6J and KPC mice was downloaded from the Sequence Read Archive (SRA) (accession PRJNA749728) [ 16 ]. The reads were aligned using STAR to the GRCm39 mouse reference gene with GENCODE M27 annotations using the nf-core/rnaseq analysis pipeline version 3.0 [ 17 ].…”

CXCR2 inhibition enables NASH-HCC immunotherapy

“…Hence, TAN-enriched immune hubs observed in AZD5069/anti-PD1 treated tumours are able to avoid ICI signals that might otherwise cause immune exhaustion. AZD5069/anti-PD1 TANs also displayed a strong correlation with transcriptional changes seen in neutrophils during an acute systemic inflammatory response, 50 including expression of genes involved in exocytosis, myeloid cell activation and degranulation ( online supplemental figure 7G, H ). Finally, these AZD5069/anti-PD1 TANs closely resembled a recently objectively characterised acute-inflammatory immature-Ly6G Int neutrophil population isolated from lipopolysaccharide-(LPS)-treated mice 50 ( online supplemental figure 7I ).…”