Effective prevention and therapy of experimental allergic asthma using a GATA-3-specific DNAzyme.
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 TH2 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.


CONCLUSIONS
These results indicate that topical application of the GATA-3-specific DNAzyme is a promising novel approach for the treatment of allergic bronchial asthma.
Sel
Serdar
S
Department of Clinical Chemistry and Molecular Diagnostics, Medical Faculty, Philipps University of Marburg, Marburg, Germany.
Wegmann
Michael
M
Dicke
Tanja
T
Sel
Sarper
S
Henke
Wolfgang
W
Yildirim
Ali O
AO
Renz
Harald
H
Garn
Holger
H
D000208:Acute Disease; D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000627:therapeutic use; D001249:Asthma / Q000188:drug therapy* / Q000201:enzymology / Q000517:prevention & control*; D016535:Bronchial Hyperreactivity / Q000201:enzymology / Q000473:pathology / Q000517:prevention & control; D045744:Cell Line, Tumor; D002908:Chronic Disease; D016373:DNA, Antisense / Q000494:pharmacology; D021881:DNA, Catalytic / Q000627:therapeutic use*; D004195:Disease Models, Animal; D004789:Enzyme Activation / Q000235:genetics / Q000276:immunology; D005260:Female; D050990:GATA3 Transcription Factor / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism*; D018836:Inflammation Mediators / Q000037:antagonists & inhibitors / Q000502:physiology; D051379:Mice; D008807:Mice, Inbred BALB C; D010047:Ovalbumin / Q000633:toxicity; D012333:RNA, Messenger / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D034741:RNA, Small Interfering / Q000494:pharmacology; D013379:Substrate Specificity / Q000235:genetics / Q000276:immunology
D000894:Anti-Inflammatory Agents, Non-Steroidal; D016373:DNA, Antisense; D021881:DNA, Catalytic; D050990:GATA3 Transcription Factor; C494748:Gata3 protein, mouse; D018836:Inflammation Mediators; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D010047:Ovalbumin