Identification of immunosuppressive neutrophils using multi-omics: why functional testing remains key

Polymorphonuclear neutrophils (PMNs) are innate cells that may act as myeloid-derived suppressor cells (MDSCs), most often characterized by their immunosuppressive capacity towards immune cells within the tumor microenvironment. Over the last decade, many studies have tried to characterize these PMN-MDSCs. Although all human neutrophils obtain suppressive activity upon activation, specialized subsets, including low-density cells, have been proposed to exert MDSC activity without requiring prior stimulation. Single-cell RNA sequencing (scRNAseq) has created a new horizon to study possible neutrophil subsets. As such, the identification of distinct neutrophil subtypes in tumors may better characterize immunosuppressive neutrophils, in the end leading to specific targeting strategies in cancer patients. Nevertheless, scRNAseq has not yet led to a clear immunophenotypic characterization of PMN-MDSCs, and clinical relevance by functional testing is still lacking. Studies on the discrepancy in RNA abundance, protein expression and functionality in neutrophils indicate that the future for defining functional neutrophil subsets will rely on further development of single-cell techniques, like proteomics, to truly carve out neutrophil subsets and plasticity.