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人支气管内膜活组织检查的转录组测序(RNA-Seq):哮喘与对照。

Transcriptome sequencing (RNA-Seq) of human endobronchial biopsies: asthma versus controls.

机构信息

Dept of Respiratory Medicine, Academic Medical Centre, Amsterdam, The Netherlands.

出版信息

Eur Respir J. 2013 Sep;42(3):662-70. doi: 10.1183/09031936.00115412. Epub 2013 Jan 11.

Abstract

The cellular and molecular pathways in asthma are highly complex. Increased understanding can be obtained by unbiased transcriptomic analysis (RNA-Seq). We hypothesised that the transcriptomic profile of whole human endobronchial biopsies differs between asthma patients and controls. First, we investigated the feasibility of obtaining RNA from whole endobronchial biopsies suitable for RNA-Seq. Secondly, we examined the difference in transcriptomic profiles between asthma and controls. This cross-sectional study compared four steroid-free atopic asthma patients and five healthy nonatopic controls. Total RNA from four biopsies per subject was prepared for RNA-Seq. Comparison of the numbers of reads per gene in asthma and controls was based on the Poisson distribution. 46 genes were differentially expressed between asthma and controls, including pendrin, periostin and BCL2. 10 gene networks were found to be involved in cellular morphology, movement and development. RNA isolated from whole human endobronchial biopsies is suitable for RNA-Seq, showing different transcriptomic profiles between asthma and controls. Novel and confirmative genes were found to be linked to asthma. These results indicate that biological processes in the airways of asthma patients are regulated differently when compared to controls, which may be relevant for the pathogenesis and treatment of the disease.

摘要

哮喘的细胞和分子途径非常复杂。通过无偏转录组分析(RNA-Seq)可以获得更深入的了解。我们假设整个人支气管活检的转录组图谱在哮喘患者和对照组之间存在差异。首先,我们研究了从适合 RNA-Seq 的整个支气管活检中获取 RNA 的可行性。其次,我们检查了哮喘和对照组之间转录组谱的差异。这项横断面研究比较了 4 名无类固醇的特应性哮喘患者和 5 名健康的非特应性对照者。每个受试者的 4 个活检标本的总 RNA 均用于 RNA-Seq。基于泊松分布比较哮喘和对照组每个基因的读数数。哮喘和对照组之间有 46 个基因表达差异,包括 pendrin、periostin 和 BCL2。发现 10 个基因网络与细胞形态、运动和发育有关。从整个人支气管活检中分离的 RNA 适合 RNA-Seq,显示哮喘和对照组之间存在不同的转录组谱。发现与哮喘相关的新的和证实的基因。这些结果表明,与对照组相比,哮喘患者气道中的生物学过程受到不同的调节,这可能与疾病的发病机制和治疗有关。

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