The Development of Adeno-Associate Viral Vectors Targeting Inflammatory Genes in Asthmatic Murine Models

Asthma is a complex pulmonary inflammatory disease. Generally, it is believed that the susceptible individuals have an abnormal immune response to inhaled allergens. According to some previous studies, Th2 cells and their cytokines play pivotal roles in the pathogenesis of asthma. In addition, the activation of the mast cells and eosinophils in the airway mucosa releases high levels of proinflammatory mediators which induce bronchial obstruction, airway hyper-responsiveness, and airway inflammation. We have established the hybrid adeno-associate virus (AAV) 2/9 vector system to carry shRNA specific to eotaxin-1 (CCL11) and results showed efficient reduction of eosinophilia, airway hyperresponsiveness (AHR) and local Th2 cytokine levels in allergen-sensitized mice. The cDNA sequence of Clara cell 10 kD protein (CC10) was also constructed and had similar therapeutic effect as AAV-shCCL11. The activity of inflammasomes and extracellular ATP have recently been reported to correlate with the pathogenesis of asthma, particular the purinergic P2X7 receptor and its related molecules, include NLRP3, Pannexin-1, and IL-33. Therefore, we will evaluate whether the expression of CD39, CD73 or ST2 by AAV vector would reduce the inflammasome activity in asthmatic mice. The AAV gene delivery systems will also target the inflammasome components, including siRNA sequences to NLRP3, P2X7 receptor, or IL-33. Further, the expression of Pannexin-1 mimetic blocking peptide (10panx1) from epithelial cells will be examined. The airway hyperresponsiveness, cell infiltration in BALF, ATP levels, and the expression of inflammasome molecules will be determined. The results will offer a better understanding of the roles of inflammasome activity and extracellular ATP in asthmatic pathogenesis. The novel therapeutic approaches with those molecules will be developed based on the information obtained from this study.

Project ID:PC10301-0301
External Project ID:NSC102-2320-B182-030-MY3