背景:慢性鼻窦炎伴鼻息肉(CRSwNP)在美国以2型炎症为特征,然而,嗜酸性粒细胞在CRSwNP中的实际作用尚不清楚.
目的:通过单细胞RNA测序(scRNA-Seq)分析鼻息肉(NP)组织中嗜酸性粒细胞的作用和异质性。
方法:从5例对照患者和5例CRSwNP患者中获得鼻窦组织(NP和对照窦组织)和患者匹配的外周血(PB)样本。在处理scRNA-Seq之前富集嗜酸性粒细胞。通过基于微孔的scRNA-Seq技术(BD-Rhapsody平台)测定嗜酸性粒细胞中的基因表达谱。我们通过基因本体论(GO)富集和途径分析预测了NP嗜酸性粒细胞的整体功能,并通过流式细胞术确认了所选基因的表达。
结果:过滤掉污染细胞后,我们从对照PB中检测到5542个嗜酸性粒细胞,来自CRSwNPPB的3,883个嗜酸性粒细胞,通过scRNA-Seq,来自对照窦组织的101个嗜酸性粒细胞(未包括在进一步分析中)和来自NP的9,727个嗜酸性粒细胞。我们发现204个基因下调,354个基因在NP嗜酸性粒细胞中上调,与所有PB嗜酸性粒细胞相比(>1.5倍,Padj<0.05)。NP嗜酸性粒细胞中上调的基因与激活有关,细胞因子介导的信号传导,生长因子活性,NF-κB信号传导和抗凋亡分子。NP嗜酸性粒细胞显示4个簇,揭示了NP组织中嗜酸性粒细胞的潜在异质性。
结论:NP组织中嗜酸性粒细胞升高似乎存在于几种亚型中,这些亚型可能在CRSwNP中起重要的致病作用,部分通过控制炎症和其他细胞的过度增殖。
BACKGROUND: 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.
OBJECTIVE: 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.
METHODS: 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.
RESULTS: 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, Padj < .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.
CONCLUSIONS: 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.