BACKGROUND 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. OBJECTIVE To reveal the roles and heterogeneity of neutrophils in NP tissue by single-cell RNA sequencing analysis. METHODS 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. RESULTS 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. CONCLUSIONS Elevated neutrophils in NP tissue appear to exist in several subphenotypes that may play important pathogenic roles in CRSwNP.

中文翻译：

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通过单细胞 RNA 测序分析来自鼻息肉的人中性粒细胞，揭示了中性粒细胞在慢性鼻-鼻窦炎中的作用。


背景 慢性鼻窦炎伴鼻息肉 （CRSwNP） 的特征是 2 型 （T2） 炎症。最近的研究，包括我们自己的研究，表明 T2 鼻息肉 （NP） 中的中性粒细胞也升高，并且升高的中性粒细胞显示出活化的表型。然而，中性粒细胞在 T2 CRSwNP 的 NP 发病机制中的实际作用在很大程度上仍不清楚。目的 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。方法 我们使用来自 5 个对照鼻窦组织、 5 个 NP 组织和患者匹配的外周血 （PB） 样本的富含粒细胞的样本开发了一种新的基于微孔的单细胞 RNA 测序测定法。这种方法允许通过 Benjamini-Hochberg 算法检查 NP 嗜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 嗜中性粒细胞的整体功能。结果在执行所有质量控制步骤后，我们成功检测到中性粒细胞。与所有 PB 中性粒细胞 （13,591 个细胞） 相比，我们在 NP 中性粒细胞 （1,151 个细胞） 中鉴定出 333 个下调和 128 个上调基因 （>1.5 倍，q < 0.05），并发现 NP 中性粒细胞与所有 PB 和对照窦组织中性粒细胞 （3,136 个细胞） 相比，NP 中性粒细胞中普遍出现失调基因。NP 中性粒细胞中通常下调的基因与先天免疫系统有关，上调的基因与核因子-κB 信号传导、细胞因子活性和细胞对含氧化合物的反应有关。NP 中性粒细胞显示 4 个簇，揭示了 NP 组织中中性粒细胞的潜在异质性。结论 NP 组织中中性粒细胞升高似乎存在于几种亚表型中，这些亚表型可能在 CRSwNP 中起重要致病作用。