Related Experiment Video
Updated: Oct 26, 2026
ATAC-Seq Library Preparation of Murine Bone Marrow-Derived Neutrophils
Published on: January 3, 2025
Single-cell RNA sequencing highlights the heterogeneity of neutrophils in granulomatosis with polyangiitis
Shuning Guo1,2,3,4, Yangzhong Zhou1,2,3,4, Dazhi Pang5
1Department of Rheumatology and Clinical Immunology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, China.
Background And Objectives:
Granulomatosis with polyangiitis (GPA), a clinical phenotype of anti-neutrophil cytoplasmic antibody-associated vasculitis, is characterized by necrotizing inflammation of blood vessels. While neutrophil dysregulation is central to its pathogenesis, its cellular heterogeneity and dynamic functional states remain poorly defined. To better understand the transcriptomic profile of neutrophils, we applied single-cell RNA sequencing (scRNA-seq) to peripheral blood samples from GPA patients and healthy controls (HCs).
Methods:
Peripheral blood samples were collected from three patients with active GPA and three HCs, followed by sequencing using the BD Rhapsody platform (Becton, Dickinson and Company). In a subsequent validation cohort, flow cytometry and quantitative real-time polymerase chain reaction were used to confirm specific neutrophil subtypes and explore their potential clinical significance.
Results:
Seven neutrophil clusters with distinct transcriptomic and functional profiles were identified. CXCL8-high and TXNIP-high subsets, predominant in HCs and GPA patients respectively, showed inflammatory profiles. CXCL8-high subsets also expressed immunoregulatory molecules to restrain inflammation. MMP8-high neutrophils showed enhanced antimicrobial activity, whereas IFIT1-high cells were linked to antiviral responses, suggesting that prior infections may contribute to anti-neutrophil cytoplasmic antibody-associated vasculitis (AAV) pathogenesis. PLA2G7-high neutrophils, identified as terminally differentiated by pseudotime analysis, were associated with disease relapse and renal involvement, indicating potential as AAV biomarkers. Transcriptional profiles of peripheral T and natural killer (NK) cells were also characterized, along with the analysis of systemic immune cell-cell communication networks.
Conclusion:
Our study enhances the understanding of the dynamic functional roles of neutrophils in GPA and may facilitate the development of therapeutic strategies aimed at selectively targeting disease-specific neutrophil subsets.
