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Abstract：
BACKGROUND: Asthma is the most common chronic disease in children with an increasing prevalence. Its development is caused by genetic and environmental factors and allergic sensitization is a known trigger. Dog allergens affect up to 30% of all children and dog dander-sensitized children show increased expression of cystatin-1 (CST1) and eotaxin-3 (CCL26) in nasal epithelium. The aim of our study was to investigate the functional mechanism of CST1 and CCL26 in the alveolar basal epithelial cell line A549.
METHODS: A549 cells were transfected with individual overexpression vectors for CST1 and CCL26 and RNA sequencing was performed to examine the transcriptomics. edgeR was used to identify differentially expressed genes (= DEG, |log2 FC | ≥ 2, FDR < 0.01). The protein expression levels of A549 cells overexpressing CST1 and CCL26 were analyzed using the Target 96 inflammation panel from OLINK (antibody-mediated proximity extension-based assay; OLINK Proteomics). Differentially expressed proteins were considered with a |log2 FC| ≥ 1, p < .05.
RESULTS: The overexpression of CST1 resulted in a total of 27 DEG (1 upregulated and 26 downregulated) and the overexpression of CCL26 in a total of 137 DEG (0 upregulated and 137 downregulated). The gene ontology enrichment analysis showed a significant downregulation of type I and III interferon signaling pathway genes as well as interferon-stimulated genes. At the protein level, overexpression of CST1 induced a significantly increased expression of CCL3, whereas CCL26 overexpression led to increased expression of HGF, and a decrease of CXCL11, CCL20, CCL3 and CXCL10.
CONCLUSIONS: Our results indicate that an overexpression of CST1 and CCL26 cause a downregulation of interferon related genes and inflammatory proteins. It might cause a higher disease susceptibility, mainly for allergic asthma, as CCL26 is an agonist for CCR-3-carrying cells, such as eosinophils and Th2 lymphocytes, mostly active in allergic asthma.
METHODS: A549 cells were transfected with individual overexpression vectors for CST1 and CCL26 and RNA sequencing was performed to examine the transcriptomics. edgeR was used to identify differentially expressed genes (= DEG, |log2 FC | ≥ 2, FDR < 0.01). The protein expression levels of A549 cells overexpressing CST1 and CCL26 were analyzed using the Target 96 inflammation panel from OLINK (antibody-mediated proximity extension-based assay; OLINK Proteomics). Differentially expressed proteins were considered with a |log2 FC| ≥ 1, p < .05.
RESULTS: The overexpression of CST1 resulted in a total of 27 DEG (1 upregulated and 26 downregulated) and the overexpression of CCL26 in a total of 137 DEG (0 upregulated and 137 downregulated). The gene ontology enrichment analysis showed a significant downregulation of type I and III interferon signaling pathway genes as well as interferon-stimulated genes. At the protein level, overexpression of CST1 induced a significantly increased expression of CCL3, whereas CCL26 overexpression led to increased expression of HGF, and a decrease of CXCL11, CCL20, CCL3 and CXCL10.
CONCLUSIONS: Our results indicate that an overexpression of CST1 and CCL26 cause a downregulation of interferon related genes and inflammatory proteins. It might cause a higher disease susceptibility, mainly for allergic asthma, as CCL26 is an agonist for CCR-3-carrying cells, such as eosinophils and Th2 lymphocytes, mostly active in allergic asthma.
摘要：
背景：哮喘是儿童中最常见的慢性疾病，患病率越来越高。其发展是由遗传和环境因素引起的,并且变应性致敏是已知的触发因素。狗过敏原影响多达30％的所有儿童，狗皮屑致敏的儿童在鼻上皮中显示出胱抑素-1（CST1）和eotaxin-3（CCL26）的表达增加。我们研究的目的是研究CST1和CCL26在肺泡基底上皮细胞系A549中的功能机制。
方法：用CST1和CCL26的单个过表达载体转染A549细胞，并进行RNA测序以检查转录组学。edgeR用于鉴定差异表达的基因(=DEG，|log2FC|≥2,FDR<0.01)。使用来自OLINK的靶96炎症组(基于抗体介导的邻近延伸的测定;OLINK蛋白质组学)分析过表达CST1和CCL26的A549细胞的蛋白质表达水平。差异表达的蛋白质被认为具有|log2FC|≥1,p<.05。
结果:CST1的过表达导致总共27°(1个上调,26个下调),并且CCL26的过表达导致总共137°(0个上调,137个下调)。基因本体富集分析显示I型和III型干扰素信号通路基因以及干扰素刺激基因显著下调。在蛋白质水平，CST1的过表达诱导CCL3的表达显著增加，而CCL26的过表达导致HGF的表达增加，CXCL11、CCL20、CCL3和CXCL10减少。
结论：我们的结果表明，CST1和CCL26的过表达导致干扰素相关基因和炎症蛋白的下调。它可能会导致更高的疾病易感性，主要用于过敏性哮喘，因为CCL26是携带CCR-3的细胞的激动剂，如嗜酸性粒细胞和Th2淋巴细胞，主要活跃于过敏性哮喘。
方法：用CST1和CCL26的单个过表达载体转染A549细胞，并进行RNA测序以检查转录组学。edgeR用于鉴定差异表达的基因(=DEG，|log2FC|≥2,FDR<0.01)。使用来自OLINK的靶96炎症组(基于抗体介导的邻近延伸的测定;OLINK蛋白质组学)分析过表达CST1和CCL26的A549细胞的蛋白质表达水平。差异表达的蛋白质被认为具有|log2FC|≥1,p<.05。
结果:CST1的过表达导致总共27°(1个上调,26个下调),并且CCL26的过表达导致总共137°(0个上调,137个下调)。基因本体富集分析显示I型和III型干扰素信号通路基因以及干扰素刺激基因显著下调。在蛋白质水平，CST1的过表达诱导CCL3的表达显著增加，而CCL26的过表达导致HGF的表达增加，CXCL11、CCL20、CCL3和CXCL10减少。
结论：我们的结果表明，CST1和CCL26的过表达导致干扰素相关基因和炎症蛋白的下调。它可能会导致更高的疾病易感性，主要用于过敏性哮喘，因为CCL26是携带CCR-3的细胞的激动剂，如嗜酸性粒细胞和Th2淋巴细胞，主要活跃于过敏性哮喘。
