Pseudotyped Adeno-Associated Virus 2/9-DeliveredCCL11shRNA Alleviates Lung Inflammation in an Allergen-Sensitized Mouse Model

Abstract

Airway infiltration by eosinophils is a major characteristic of chronic asthma. CCL11 (eotaxin-1) is secreted by lung epithelial cells and functions as the major chemokine for eosinophil recruitment. Pseudotyped adenoassociated virus (AAV) 2/9, composed by the AAV2 rep and AAV9 cap genes, can efficiently target lung epithelial cells and might carry gene sequences with therapeutic potential for asthma. This study aimed to determine whether pseudotyped AAV2/9 virus carrying the small hairpin RNA targeting CCL11 and expressed by CMV/U6 promoter could reduce eosinophilia and asthmatic responses in mite allergen-sensitized mice. Mice were sensitized by intraperitoneal and challenged by intratracheal injection with recombinant Dermatophagoides pteronyssinus group 2 allergen (rDp2). AAV2/9 viral vectors were intratracheally injected three days before the first challenge. AAV2/9 sh47 virus significantly reduced airway hyperresponsiveness, airway resistance, CCL11 levels, and eosinophilia in the lungs of sensitized mice. Th2 cytokines, including interleukins (IL)-4, IL-5, and IL-10, were also significantly reduced in the bronchoalveolar lavage fluid of AAV2/9 sh47 virus-treated mice. Th2 cytokine levels were also reduced in rDp2-stimulated mediastinal lymphocytes in treated mice. However, serum levels of rDp2-specific IgG1 and IgE, as well as Th2 cytokine levels in rDp2-stimulated splenocyte culture supernatants, were comparable to the sensitized control group. The results suggest that AAV2/9 sh47 virus relieved local instead of systemic inflammatory responses. Therefore, the CMV/U6 promoter with AAV2/9 viral vector, which is preferable to target lung epithelia cells, might be applied as a novel therapeutic approach for asthma.

Berberine Inhibits Pro-inflammatory Cytokine-induced IL-6 and CCL11 Production via Modulation of STAT6 Pathway in Human Bronchial Epithelial Cells

“…Significant increase of CCL11 levels is detected in blood, sputum, and exhaled breath condensate (EBC) of asthmatic patients compared with healthy subjects [9]. Moreover, the disruption of CCL11 gene expression effectively reduces tissue eosinophilia [10]. Therefore, the reduction of eosinophil recruitment by inhibiting CCL11 expression in the lungs might have therapeutic potential for asthma.…”

Sarcolipin overexpression impairs myogenic differentiation in Duchenne muscular dystrophy

“…Studies have shown that maintaining the SR Ca 2ϩ content is an important modulator of Ca i 2ϩ levels (36,53) and muscle differentiation (17,45,47). In dystrophic muscles, abnormal Ca 2ϩ cycling through sarcolemma and SR increases the Ca i 2ϩ load, an important stimulus that triggers pathological events such as activation of Ca 2ϩ -dependent proteases, improper muscle regeneration, and necrosis (1,2,8,22,57). Among several mechanisms that are responsible for Ca 2ϩ dysregulation, improving the SR Ca 2ϩ -ATPase (SERCA) function, which restores Ca 2ϩ to the SR during excitation-contraction coupling, is found to be highly effective in normalizing the Ca i 2ϩ levels and mitigating the muscle disease in DMD (18,19,32,49).…”

Intratracheal Administration of siRNA Dry Powder Targeting Vascular Endothelial Growth Factor Inhibits Lung Tumor Growth in Mice

“…Because nucleic acids are easily degraded in vivo , an appropriate vector is required for their effective delivery to targeted cells. Viral vectors, 23 , 24 lipid vectors, 25 , 26 and cationic polymers 27 , 28 have been reported as appropriate vectors for inhalable siRNA. Among them, we focused on water-soluble chitosan because of its low toxicity.…”