Comprehensive Gene Expression Signature Using RNA-Seq in Airways of Mouse Model of Severe Asthma with Fungal Sensitization
Comprehensive Gene Expression Signature Using RNA-Seq in Airways of Mouse Model of Severe Asthma with Fungal Sensitization
复制标题
DOI：
10.1159/000519172
复制
发表时间：
2021-09-29
影响因子：
2.8
通讯作者：
Sagara, Hironori
中科院分区：
文献类型：
--
作者：
Inoue, Hideki;Akimoto, Kaho;Sagara, Hironori
简介：吸入真菌过敏原可导致气道炎症和过度粘液分泌后的气道上皮损伤，这可导致伴有真菌致敏的严重哮喘（SAFS）。尚未对暴露于链格孢菌的小鼠气道（SAFS模型）进行全面的基因表达分析。方法：BALB/c小鼠经鼻给予链格孢菌提取物或磷酸盐缓冲液，每周2次，连续6周。肺切片和支气管肺泡灌洗液，以评估气道炎症。在中央气道中进行RNA-Seq，并进行基因本体（GO）分析和基因集富集分析（GSEA）以进行通路分析。使用人气道上皮细胞16 HBE 140-进行体外实验以验证RNA-Seq发现。结果：链格孢菌染毒小鼠出现嗜酸性粒细胞性气道炎症，并伴有气道黏液过度分泌和气道重塑。RNA-Seq分析显示，在链格孢菌暴露小鼠的气道中有403个上调和108个下调基因。在GO分析中，免疫球蛋白（IG）受体结合、IG产生、炎症反应和T细胞活化的功能上调，而角化和对其他生物体的防御反应的功能下调。GSEA显示阳性富集的T细胞受体复合物，免疫突触，抗原结合，肥大细胞活化，和IG受体结合，和负富集角化和角化在链格孢暴露小鼠相对于对照。链格孢菌暴露于16 HBE 14 o-细胞验证了上皮角化相关基因的下调，包括SPRR 1A、SPRR 1B和KRT 6 B。结论：RNA-Seq分析表明，链格孢菌暴露诱导小鼠气道上皮细胞的炎症反应和防御机制受损，这可能是SAFS的治疗靶点。
Introduction: Inhalation of fungal allergens induces airway epithelial damage following airway inflammation and excessive mucus secretion, which can lead to severe asthma with fungal sensitization (SAFS). Comprehensive gene expression analysis in Alternaria-exposed mouse airways, a model of SAFS, has not been conducted. Methods: BALB/c mice received intranasal administration of Alternaria extract or phosphate-buffered saline twice a week for 6 weeks. Lung sections and bronchoalveolar lavage fluid were obtained to assess airway inflammation. RNA-Seq in the central airway was performed, and gene ontology (GO) analysis and gene set enrichment analysis (GSEA) were conducted for pathway analyses. An in vitro experiment using human airway epithelial cell 16HBE14o- was performed to validate the RNA-Seq findings. Results: Eosinophilic airway inflammation with mucus overproduction and airway remodeling was observed in mice exposed to Alternaria. RNA-Seq analysis revealed 403 upregulated and 108 downregulated genes in airways of Alternaria-exposed mice. In GO analysis, the functions of immunoglobulin (Ig) receptor binding, Ig production, inflammatory response, and T-cell activation were upregulated, while those of keratinization and defense response to other organisms were downregulated. GSEA revealed positive enrichment in T-cell receptor complex, immunological synapse, antigen binding, mast cell activation, and Ig receptor binding, and negative enrichment in keratinization and cornification in Alternaria-exposed mice relative to control. Alternaria exposure to 16HBE14o- cells validated the downregulation of epithelial keratinization-related genes, including SPRR1A, SPRR1B, and KRT6B. Conclusion: RNA-Seq analysis showed that Alternaria exposure induced inflammatory response and impaired defense mechanisms in mice airway epithelium, which might be therapeutic targets for SAFS.