University Medical Center Schleswig-Holstein, Department of Pediatric Pneumology & Allergology, Campus Lübeck, Airway Research Center North (ARCN), Member of the German Center of Lung Research (DZL), Lübeck, Germany.
University of Lübeck, Institute for Medical Biometry and Statistics, Airway Research Center North (ARCN), Member of the German Center of Lung Research (DZL), Lübeck, Germany.
PLoS One. 2018 Nov 28;13(11):e0205275. doi: 10.1371/journal.pone.0205275. eCollection 2018.
Human rhinovirus infection (HRVI) plays an important role in asthma exacerbations and is thought to be involved in asthma development during early childhood. We hypothesized that HRVI causes differential DNA methylation and subsequently differential mRNA expression in epithelial cells of children with asthma. Primary nasal epithelial cells from children with (n = 10) and without (n = 10) asthma were cultivated up to passage two and infected with Rhinovirus-16 (RV-16). HRVI-induced genome-wide differences of DNA methylation in asthmatics (vs. controls) and resulting mRNA expression were analyzed by the HumanMethylation450 BeadChip Kit (Illumina) and RNA sequencing. These results were further verified by pyrosequencing and quantitative PCR, respectively. 471 CpGs belonging to 268 genes were identified to have HRVI-induced asthma-specifically modified DNA methylation and mRNA expression. A minimum-change criteria was applied to restrict assessment of genes with changes in DNA methylation and mRNA expression of at least 3% and least 0.1 reads/kb per million mapped reads, respectively. Using this approach we identified 16 CpGs, including HLA-B-associated transcript 3 (BAT3) and Neuraminidase 1 (NEU1), involved in host immune response against HRVI. HRVI in nasal epithelial cells leads to specific modifications of DNA methylation with altered mRNA expression in children with asthma. The HRVI-induced alterations in DNA methylation occurred in genes involved in the host immune response against viral infections and asthma pathogenesis. The findings of our pilot study may partially explain how HRVI contribute to the persistence and progression of asthma, and aid to identify possible new therapeutic targets. The promising findings of this pilot study would benefit from replication in a larger cohort.
人类鼻病毒感染（HRVI）在哮喘加重中起重要作用，并且被认为参与儿童早期哮喘的发展。我们假设 HRVI 导致哮喘儿童的上皮细胞中 DNA 甲基化的差异，并随后导致 mRNA 表达的差异。从患有哮喘（n = 10）和不患有哮喘（n = 10）的儿童中培养原代鼻上皮细胞，直至第 2 代，并感染鼻病毒-16（RV-16）。通过 HumanMethylation450 BeadChip Kit（Illumina）和 RNA 测序分析 HRVI 诱导的哮喘患者（与对照组相比）和随后的 mRNA 表达的全基因组 DNA 甲基化差异。通过焦磷酸测序和定量 PCR 分别进一步验证了这些结果。确定了 471 个 CpG，属于 268 个基因，这些基因具有 HRVI 诱导的哮喘特异性修饰的 DNA 甲基化和 mRNA 表达。应用最小变化标准来限制对 DNA 甲基化和 mRNA 表达发生至少 3%和每百万映射读数至少 0.1 个读数/ kb 变化的基因的评估。使用这种方法，我们鉴定了 16 个 CpG，包括 HLA-B 相关转录物 3（BAT3）和神经氨酸酶 1（NEU1），它们参与针对 HRVI 的宿主免疫反应。鼻上皮细胞中的 HRVI 导致哮喘儿童中 DNA 甲基化的特异性修饰，并改变 mRNA 表达。HRVI 诱导的 DNA 甲基化改变发生在参与宿主抗病毒感染和哮喘发病机制的免疫反应的基因中。该初步研究的发现部分解释了 HRVI 如何促进哮喘的持续和进展，并有助于确定可能的新治疗靶标。这项初步研究的有希望的发现将受益于在更大的队列中复制。
