A neutrophil-specific GSR integrates pattern recognition receptor in pathogenesis of neonatal late-onset sepsis: evidence from artificial intelligence-driven multi-omics and clinical validation

Abstract Background Neonatal late-onset sepsis (NS) is a life-threatening condition driven by a dysregulated host response to infection, where neutrophil dysfunction plays a pivotal role. Pattern recognition receptors (PRRs), as the frontline sensors of the innate immune system, are increasingly recognized as key regulators of immune cell function, yet their specific connection to neutrophil biology in NS remains largely undefined. Objective This study aimed to decode a novel PRR-neutrophil (PRRN)-associated molecular signature in NS. Methods Bulk transcriptomic data from NS peripheral samples (GSE138712) were analyzed using Limma, WGCNA, and CIBERSORT to identify shared PRRN-related differentially expressed genes (DEGs). Hub genes were narrowed down via PPI network analysis. A diagnostic model was constructed using 132 machine learning algorithms on the training set (GSE69686, NS peripheral blood bulk profile) and validated on an independent set (GSE25504, NS peripheral blood bulk profile) based on PRRN-associated hub genes. SHAP analysis identified the central hub gene in training set. Single-cell RNA sequencing data of sepsis patient peripheral blood(GSE167363) were used to validate its cell-specific distribution, functional trajectory, and regulatory network at spatial and temporal manners via cutting-edge analytical framework. AI-based drug screening (DrugReflector) and molecular docking computationally nominated a candidate therapeutic compound for NS targeting PRRN-associated central hub genes. Finally, NS patient clinical samples were collected for the validation of expression patterns of hub genes. Results We identified PRRN-associated diagnostic and candidate therapeutic models for NS patients and nominated GSR as the PRRN-associated and up-regulated hub gene in NS pathogenesis. Single-cell analysis confirmed that GSR is predominantly expressed in neutrophils within the NS samples and is enriched in pathways related to innate immune response. Conclusion This study systematically defines novel PRRN molecular co-expression patterns in NS, establishing GSR as a neutrophil-centric hub gene. These findings offer a potential diagnostic framework and an exploratory therapeutic target for NS patients, and the candidate compound requires further experimental validation.