Laboratory of Molecular and Cell Biology, Department of Pediatric Pulmonology, Allergy and Clinical Immunology, Poznan University of Medical Sciences, Poznan, Poland.
Department of Psychiatric Genetics, Poznan University of Medical Sciences, Poznan, Poland.
Clin Respir J. 2020 Dec;14(12):1191-1200. doi: 10.1111/crj.13260. Epub 2020 Sep 1.
Bronchial asthma is a chronic respiratory disease characterized by airway inflammation, allergen-induced hypersensitivity and dyspnea. Most asthmatic patients demonstrate oscillations of disease symptoms within 24 hours regulated by circadian clock genes. We hypothesized that these genes may be regulators of childhood asthma risk.
The aim was to investigate whether single-nucleotide polymorphisms (SNPs) in the circadian clock genes are associated with childhood asthma risk. We also aimed to analyze the mRNA level of clock genes in the blood of asthmatic children and NHBE cells stimulated with IL-13.
Peripheral blood was collected from 165 asthmatic and 138 healthy Polish children. NHBE cells were culture at the air-liquid interface (ALI) with IL-13 as an in vitro model of allergic inflammation. Using TaqMan probes, we genotyped 32 SNPs in: CLOCK, BMAL1, PER3 and TIMELESS. Expression analysis for TIMELESS was performed using real-time PCR with SYBR Green. For haplotype and genotype statistical analysis we used Haploview 4.2 and STATISTICA version 12, respectively. Gene expression analysis was performed in DataAssist v3.01.
We found that three polymorphisms in TIMELESS (rs2291739, rs10876890, rs11171856) and two haplotypes (TTTT and CTAC) were associated with asthma risk. We also found significantly decreased expression of TIMELESS in the blood of asthmatic children as compared to the healthy children (P = 0.0289) and in NHBE cells stimulated with IL-13 (P = 0.0302).
In our study, we showed for the first time that TIMELESS variants and expression may be associated with childhood asthma.
支气管哮喘是一种慢性呼吸道疾病，其特征为气道炎症、过敏原诱导的过敏反应和呼吸困难。大多数哮喘患者的疾病症状在 24 小时内呈波动变化，由生物钟基因调控。我们假设这些基因可能是儿童哮喘风险的调节因子。
本研究旨在探讨生物钟基因中的单核苷酸多态性（SNP）是否与儿童哮喘风险相关。我们还旨在分析哮喘儿童血液中的时钟基因 mRNA 水平以及受白介素-13（IL-13）刺激的 NHBE 细胞中的时钟基因 mRNA 水平。
从 165 例哮喘患儿和 138 例健康波兰儿童中采集外周血。NHBE 细胞在气液界面（ALI）培养，并用 IL-13 作为过敏性炎症的体外模型。使用 TaqMan 探针，我们对 CLOCK、BMAL1、PER3 和 TIMELESS 中的 32 个 SNP 进行了基因分型。使用实时 PCR 结合 SYBR Green 对 TIMELESS 的表达分析。分别使用 Haploview 4.2 和 STATISTICA 版本 12 进行 haplotype 和基因型统计分析。使用 DataAssist v3.01 进行基因表达分析。
我们发现，TIMELESS 中的三个 SNP（rs2291739、rs10876890、rs11171856）和两个单倍型（TTTT 和 CTAC）与哮喘风险相关。我们还发现，与健康儿童相比，哮喘儿童血液中的 TIMELESS 表达显著降低（P=0.0289），并且在受 IL-13 刺激的 NHBE 细胞中表达也显著降低（P=0.0302）。
在我们的研究中，我们首次表明 TIMELESS 变体和表达可能与儿童哮喘有关。
