Circulating Low Density Neutrophils In Advanced Lung Cancer Patients Exhibit Unique Immune Signatures and affect prognosis

Abstract The role of innate immunity, in especially neutrophils, in supporting anti-tumor immunity in lung cancer is understudied. Neutrophil "flavors", such as low-density neutrophils (LDN) and high-density neutrophils (HDNs), have been described in cancer patients and associated with tumor-supportive properties. Nevertheless, HDN and LDN phenotypic properties and their cancer-driven modulations are still largely unknown. Here, we aimed to evaluate the clinical relevance and significance of circulating LDN in lung cancer patients, and by using data-driven artificial intelligence supported analysis tools evaluated phenotypic states of circulating neutrophils subsets during lung cancer progression. COPD patients and healthy volunteers served as controls. Using mass cytometry (CyTOF), we aimed to identify major sub-populations within circulating LDN and HDN subsets and determine a possible change in phenotype due to cancer. LDN were highly enriched in advanced lung cancer patients (12.9±2.4%), but not in early stage patients (1.2±0.2%), healthy individuals (1.3±1.1%), or stable COPD patients (2.2±0.6%). Elevated LDN (>10% of LDF) showed a remarkable correlation with poorer prognosis in advanced patients. Our CyTOF analysis revealed the presence of three main neutrophil subsets with a great variability in the presence of CD10+ vs. CD10− LDNs between patients. We further demonstrated that both HDN and LDN retain a degree of inherent spontaneous plasticity. Deep characterization of cancer-related circulating neutrophils, as well as their modulation along tumor progression, will assist in suggesting new biomarkers and novel targets to improve response to current conventional therapies and immunotherapies.