Department of Medicine, Division of Allergy and Immunology, Chicago, Ill.
Department of Otolaryngology, Northwestern University Feinberg School of Medicine, Chicago, Ill.
J Allergy Clin Immunol. 2024 Oct;154(4):952-964. doi: 10.1016/j.jaci.2024.05.014. Epub 2024 May 24.
Chronic rhinosinusitis with nasal polyps (CRSwNP) is characterized by type 2 inflammation in the United States, but the actual roles that eosinophils play in CRSwNP remain largely unclear.
To reveal the roles and heterogeneity of eosinophils in nasal polyp (NP) tissue, we performed single cell RNA sequencing (scRNA-Seq) analysis of NP tissue.
Sinonasal tissues (NP and control sinus tissue) and patient matched peripheral blood (PB) samples were obtained from 5 control patients and 5 patients with CRSwNP. Eosinophils were enriched before processing for scRNA-Seq. The gene expression profiles in eosinophils were determined by microwell-based scRNA-Seq technology (BD Rhapsody platform). We predicted the overall function of NP eosinophils by Gene Ontology (geneontology.org) enrichment and pathway analyses and confirmed expression of selected genes by flow cytometry.
After filtering out contaminating cells, we detected 5,542 eosinophils from control PB, 3,883 eosinophils from CRSwNP PB, 101 eosinophils from control sinus tissues (not included in further analyses), and 9,727 eosinophils from NPs by scRNA-Seq. We found that 204 genes were downregulated and 354 genes upregulated in NP eosinophils compared to all PB eosinophils (>1.5-fold, P < .05). Upregulated genes in NP eosinophils were associated with activation, cytokine-mediated signaling, growth factor activity, NF-κB signaling, and antiapoptotic molecules. NP eosinophils displayed 4 clusters revealing potential heterogeneity of eosinophils in NP tissue.
Elevated eosinophils in NP tissue appear to exist in several subtypes that may play important pathogenic roles in CRSwNP, in part by controlling inflammation and hyperproliferation of other cells.
美国慢性鼻-鼻窦炎伴鼻息肉（CRSwNP）的特征为 2 型炎症，但嗜酸性粒细胞在 CRSwNP 中的实际作用在很大程度上仍不清楚。
为了揭示鼻息肉（NP）组织中嗜酸性粒细胞的作用和异质性，我们对 NP 组织进行了单细胞 RNA 测序（scRNA-Seq）分析。
从 5 例对照患者和 5 例 CRSwNP 患者中获取鼻鼻窦组织（NP 和对照鼻窦组织）和患者匹配的外周血（PB）样本。在进行 scRNA-Seq 处理之前，富集嗜酸性粒细胞。通过微孔 scRNA-Seq 技术（BD Rhapsody 平台）确定嗜酸性粒细胞的基因表达谱。我们通过基因本体论（geneontology.org）富集和途径分析预测 NP 嗜酸性粒细胞的总体功能，并通过流式细胞术确认选定基因的表达。
在剔除污染细胞后，我们通过 scRNA-Seq 从对照 PB 中检测到 5542 个嗜酸性粒细胞，从 CRSwNP PB 中检测到 3883 个嗜酸性粒细胞，从对照鼻窦组织中检测到 101 个嗜酸性粒细胞（未纳入进一步分析），从 NP 中检测到 9727 个嗜酸性粒细胞。与所有 PB 嗜酸性粒细胞相比（>1.5 倍，P<.05），NP 嗜酸性粒细胞中 204 个基因下调，354 个基因上调。NP 嗜酸性粒细胞中上调的基因与激活、细胞因子介导的信号转导、生长因子活性、NF-κB 信号转导和抗凋亡分子有关。NP 嗜酸性粒细胞显示出 4 个簇，揭示了 NP 组织中嗜酸性粒细胞的潜在异质性。
NP 组织中升高的嗜酸性粒细胞似乎存在几种亚型，这些亚型可能通过控制炎症和其他细胞的过度增殖，在 CRSwNP 中发挥重要的致病作用。
