Single-cell RNA sequencing reveals neutrophil differentiation-related genes for immunotherapy response prediction in colorectal cancer

Immunotherapy has demonstrated promising results in treating colorectal cancer (CRC), yet not all patients benefit equally, highlighting the need for predictive biomarkers and additional adjuvant. Neutrophils, pivotal in immune regulation and inflammation, remain underexplored in the context of CRC immunotherapy, holding the potential targets for improving immunotherapy efficacy. Single-cell RNA sequencing data from 19 CRC patients, including immunotherapy-treated individuals and controls, were analyzed. Genes associated with neutrophil differentiation, termed Neutrophil Differentiation-Related Genes (NDRGs), were identified through trajectory analysis and refined to nine genes (TMBIM6, CTSS, CYCS, DDX3X, DYNLL1, LGALS1, GANI2, RPS29, and TUBA1A) using Elastic Net. Drug screening was performed to identify potential therapeutic agents targeting NDRGs. Immunotherapy induced a shift in neutrophil composition, marked by a reduction in inflammatory neutrophils and an increase in immune neutrophils. The nine NDRGs were successfully validated and utilized to develop a predictive model for immunotherapy response. Drug screening identified Ivermectin as a potential therapeutic agent targeting NDRGs. This study highlights the dynamic role of neutrophils in the CRC immune microenvironment, their potential as biomarkers for immunotherapy response, and their utility as therapeutic targets to improve immunotherapy outcomes.