Gene Expression Profiles of Acute Exacerbations of Idiopathic Pulmonary Fibrosis
Gene Expression Profiles of Acute Exacerbations of Idiopathic Pulmonary Fibrosis
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DOI：
10.1164/rccm.200810-1596oc
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发表时间：
2009-07-15
影响因子：
24.7
通讯作者：
Kaminski, Naftali
中科院分区：
文献类型：
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作者：
Konishi, Kazuhisa;Gibson, Kevin F.;Kaminski, Naftali
基本原理。特发性肺纤维化（IPF）急性加重的分子机制尚不清楚。我们研究了IPF急性加重的全局基因表达特征。目的：了解IPF急性加重期的基因表达模式。方法：从23个IPF稳定肺、8个IPF急性加重肺（IPF- aex）和15个对照肺中提取RNA，在Agilent基因表达芯片上进行杂交。利用Spotfire和Genomica对基因进行功能分析。MMP1、MMP7、AGER、DEFA1-3、COL1A2和CCNA2的基因验证采用实时定量逆转录聚合酶链反应进行。免疫组织化学和原位末端脱氧核苷酸转移酶dUTP缺口末端标记分析在用于微阵列的相同组织上进行。对对照组、稳定型IPF患者和IPF- aex患者的血浆进行α -防御素ELISA检测。测量和主要结果：IPF- aex和IPF样品中区分IPF与对照肺的基因表达模式相似。579个基因在稳定IPF和IPF- aex之间存在差异表达（错误发现率< 5%）。这些基因的功能分析没有显示感染性或压倒性炎症病因的任何证据。CCNA2和α -防御素是上调最多的基因。CCNA2和α -防御素蛋白水平也较高，并定位于IPF-AEx上皮，也检测到广泛的细胞凋亡。IPF-AEx患者外周血α -防御素水平升高。结论：我们的研究结果表明，IPF-AEx具有增强上皮损伤和增殖的特征，其表现为CCNA2和α -防御素的增加以及上皮细胞的凋亡。外周血和肺中α -防御素的增加可能表明它们可以作为这种疾病的生物标志物。
Rationale. The molecular mechanisms underlying acute exacerbations of idiopathic pulmonary fibrosis (IPF) are poorly understood. We studied the global gene expression signature of acute exacerbations of IPF.Objectives: To understand the gene expression patterns of acute exacerbations of IPF.Methods: RNA was extracted from 23 stable IPF lungs, 8 IPF lungs with acute exacerbation (IPF-AEx), and 15 control lungs and used for hybridization on Agilent gene expression microarrays. Functional analysis of genes was performed with Spotfire and Genomica. Gene validations for MMP1, MMP7, AGER, DEFA1-3, COL1A2, and CCNA2 were performed by real-time quantitative reverse transcription-polymerase chain reaction. Immunohistochemistry and in situ terminal deoxynucleotidyltransferase dUTP nick end-labeling assays were performed on the same tissues used for the microarray. ELISA for alpha-defensins was performed on plasma from control subjects, patients with stable IPF, and patients with IPF-AEx.Measurements and Main Results: Gene expression patterns in IPF-AEx and IPF samples were similar for the genes that distinguish IPF from control lungs. Five hundred and seventy-nine genes were differentially expressed (false discovery rate < 5%) between stable IPF and IPF-AEx. Functional analysis of these genes did not indicate any evidence of an infectious or overwhelming inflammatory etiology. CCNA2 and alpha-defensins were among the most up-regulated genes. CCNA2 and alpha-defensin protein levels were also higher and localized to the epithelium of IPF-AEx, where widespread apoptosis was also detected. alpha-Defensin protein levels were increased in the peripheral blood of patients with IPF-AEx.Conclusions: Our results indicate that IPF-AEx is characterized by enhanced epithelial injury and proliferation, as reflected by increases in CCNA2 and alpha-defensins and apoptosis of epithelium. The concomitant increase in alpha-defensins in the peripheral blood and lungs may suggest their use as biomarkers for this disorder.