Single cell RNA profiling of neutrophils in a model of pancreatic orthotopic cancer

Neutrophils are increasingly recognized as key players in the tumour immune response, and are associated with poor clinical outcomes. Despite recent advances characterizing the diversity of neutrophil states in cancer, common trajectories and mechanisms governing the ontogeny and relationship between these neutrophil states remain undefined. Single-cell RNA sequencing (scRNAseq) of neutrophils from the bone marrow, spleen, and blood in addtion to the pancreatic tumor thus uncovers an unique tumor differentiation trajectory, where immature and mature neutrophils converge into a terminally differentiated tumor neutrophil state. CD11b+CD115-Ly6G+ neutrophils were sorted by fluorescence activated cell sorting (FACS) from the bone marrow, spleen, blood and tumor from mice bearing orthotopic tumors previously established from the Pdx1Cre; KrasG12D/+; Trp53R172H/+ (KPC) genetically modified mouse model (n=2). Each sample was independently labelled with TotalSeq-A hashtag antibodies (A0301-A0308), before pooling for 10X v3 3' scRNAseq.