Effective prevention and therapy of experimental allergic asthma using a GATA-3-specific DNAzyme
Effective prevention and therapy of experimental allergic asthma using a GATA-3-specific DNAzyme
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DOI：
10.1016/j.jaci.2007.12.1175
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发表时间：
2008-04-01
影响因子：
14.2
通讯作者：
Garn, Holger
中科院分区：
文献类型：
--
作者：
Sel, Serdar;Wegmann, Michael;Garn, Holger
背景：过敏性支气管哮喘是一种慢性气道炎症性疾病。转录因子GATA-3被证明在T(H)2细胞活化中发挥重要作用，但也在调节支气管哮喘涉及的其他细胞类型，包括肥大细胞、嗜酸性粒细胞和上皮细胞。DNAzymes是一类新的反义分子，它结合了DNA碱基配对的特异性和固有的rna切割酶活性。目的：制备GATA-3 mrna特异性DNAzyme并分析其对实验性过敏性哮喘小鼠模型的预防作用。方法：采用体外裂解法筛选活性最高的DNAzyme（定名为gd21）。通过支气管肺泡灌洗的炎症细胞和细胞因子分析来评估变应性气道炎症。肺组织，包括杯状细胞增生和肺功能，用头部体积脉搏波分析。结果：在急性变应性气道炎症模型中，经鼻给药gd21可预防气道炎症和粘液产生，抑制气道对甲胆碱的高反应性发展。在慢性实验性哮喘模型中也发现了类似的效果。有趣的是，gd21至少与其他反义分子一样有效，并且未检测到脱靶效应。进一步的实验表明，肺表面活性剂可能作为内源性转染剂促进gd21的细胞摄取。结论：gata -3特异性DNAzyme外用治疗变应性支气管哮喘是一种有前景的新方法。
Background: Allergic bronchial asthma is a chronic inflammatory disease of the airways. The transcription factor GATA-3 was shown to play an important role in T(H)2 cell activation, but also in the regulation of other cell types involved in bronchial asthma including mast cells, eosinophils, and epithelial cells. DNAzymes represent a new class of antisense molecules that combines the specificity of DNA base pairing with an inherent RNA-cleaving enzymatic activity.Objective: To develop a GATA-3 mRNA-specific DNAzyme and analyze its allergy-preventing activity in murine models of experimental allergic asthma.Methods: The most active DNAzyme (termed gd21) was selected by in vitro cleavage assays. Allergic airway inflammation was assessed by inflammatory cell and cytokine analysis within bronchoalveolar lavage. Lung histology, including goblet cell hyperplasia and lung function, was analyzed using head-out body-plethysmography.Results: Intranasal administration of gd21 prevented airway inflammation and mucus production and inhibited development of airway hyperresponsiveness to methacholine in models of acute allergic airway inflammation. Similar effects were also detected in a model of chronic experimental asthma. Interestingly, gd21 was at least as effective as other antisense molecules, and off-target effects were not detected. Further experiments indicated that pulmonary surfactant may facilitate the cellular uptake of gd21 by acting as an endogenous transfectant.Conclusion: These results indicate that topical application of the GATA-3-specific DNAzyme is a promising novel approach for the treatment of allergic bronchial asthma.