Iwasaki Naruhito, Poposki Julie A, Kidoguchi Masanori, Oka Aiko, Klingler Aiko I, Stevens Whitney W, Suh Lydia A, Bai Junqin, Peters Anju T, Grammer Leslie C, Welch Kevin C, Smith Stephanie S, Conley David B, Bochner Bruce S, Schleimer Robert P, Kern Robert C, Tan Bruce K, Kato Atsushi
Division of Allergy and Immunology, Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, Ill.
Department of Otolaryngology, Northwestern University Feinberg School of Medicine, Chicago, Ill.
J Allergy Clin Immunol. 2025 Mar;155(3):843-855. doi: 10.1016/j.jaci.2024.10.032. Epub 2024 Nov 9.
Chronic rhinosinusitis with nasal polyps (CRSwNP) is characterized by type 2 (T2) inflammation. Recent studies, including our own, suggest that neutrophils are also elevated in T2 nasal polyps (NP) and that elevated neutrophils display an activated phenotype. However, the actual roles of neutrophils in NP pathogenesis in T2 CRSwNP are still largely unclear.
To reveal the roles and heterogeneity of neutrophils in NP tissue by single-cell RNA sequencing analysis.
We developed a novel microwell-based single-cell RNA sequencing assay using granulocyte-enriched samples from 5 control sinus tissues, 5 NP tissues and patient-matched peripheral blood (PB) samples. This approach allowed for examination of differential expression of genes in NP neutrophils by the Benjamini-Hochberg algorithm and predicted the overall function of NP neutrophils by pathway and Gene Ontology enrichment analyses.
After performing all quality control steps, we successfully detected neutrophils. We identified 333 downregulated and 128 upregulated genes in NP neutrophils (1,151 cells) compared with all PB neutrophils (13,591 cells) (>1.5-fold, q < 0.05) and found commonly dysregulated genes in NP neutrophils compared with both all PB and control sinus tissue neutrophils (3,136 cells). Commonly downregulated genes in NP neutrophils were associated with the innate immune system, and upregulated genes were associated with nuclear factor-κB signaling, cytokine activity, and cellular response to oxygen-containing compounds. NP neutrophils displayed 4 clusters revealing potential heterogeneity of neutrophils in NP tissue.
Elevated neutrophils in NP tissue appear to exist in several subphenotypes that may play important pathogenic roles in CRSwNP.
伴鼻息肉的慢性鼻-鼻窦炎（CRSwNP）以2型（T2）炎症为特征。包括我们自己的研究在内的近期研究表明，T2型鼻息肉（NP）中中性粒细胞也增多，且增多的中性粒细胞表现出活化表型。然而，在T2型CRSwNP中，中性粒细胞在NP发病机制中的实际作用仍不清楚。
通过单细胞RNA测序分析揭示中性粒细胞在NP组织中的作用和异质性。
我们开发了一种基于微孔的新型单细胞RNA测序检测方法，使用来自5个对照鼻窦组织、5个NP组织和患者匹配的外周血（PB）样本的粒细胞富集样本。该方法通过Benjamini-Hochberg算法检测NP中性粒细胞中基因的差异表达，并通过通路和基因本体富集分析预测NP中性粒细胞的整体功能。
在执行所有质量控制步骤后，我们成功检测到了中性粒细胞。与所有PB中性粒细胞（13591个细胞）相比，我们在NP中性粒细胞（1151个细胞）中鉴定出333个下调基因和128个上调基因（>1.5倍，q<0.05），并发现与所有PB和对照鼻窦组织中性粒细胞（3136个细胞）相比，NP中性粒细胞中存在常见的失调基因。NP中性粒细胞中常见的下调基因与固有免疫系统相关，上调基因与核因子-κB信号传导、细胞因子活性和对含氧化合物的细胞反应相关。NP中性粒细胞表现出4个簇，揭示了NP组织中中性粒细胞的潜在异质性。
NP组织中增多的中性粒细胞似乎存在几种亚表型，可能在CRSwNP中发挥重要的致病作用。
