AAV vectors based on serotype 2 have been evaluated extensively for lung-directed gene transfer in the treatment of cystic fibrosis (CF). Low levels of gene transfer, however, have limited the utility of AAV serotype 2 vectors in this application. Higher levels of transduction have indeed been achieved with AAV vectors based on the other known serotypes 1 and 5 indicating that entry may be a barrier to efficient gene transfer, in an attempt to further assess the potential of AAV for gene therapy of CF, we undertook a comprehensive analysis of human and nonhuman primate tissue for the presence of endogenous AAV. The goals of these studies were to further understand the biology of natural AAV infections as well as to identifydifferent AAV that would yield vectors with improved gene transfer to lung. This recently completed survey revealed endogenous AAV sequences in almost 20% of all tissues screened. Characterization of these sequences demonstrated substantial structural, serologic, and functional heterogeneity. Phylogenetic analysis demonstrated the presence of related groups of AAVs which we call clades. These data provide a basis for identifying key elements of AAV biology relevant to gene therapy as well as to create potentially improved vectors.
The Specific Aims are as follows. 1) Develop AAV vectors utilizing capsid sequences from the novel isolates. Preliminary studies indicate an extremely high level of gene transfer to lung utilizingmembers of the AAV8 and 9 clades. These will be further characterized with respect to in vivo gene transfer in both mice and nonhuman primates as well as for serologic properties. 2) The basis for the enhanced gene transfer with novel AAVs will be further evaluated. An attempt will be made to identify the receptors to at least two of the new AAVs and to evaluate the tissue distribution of these novel receptors. 3) A number of approaches will be undertaken to improve the utility of the novel AAV vectors. We will develop better methods of purification, evaluate the role of double copy vectors to overcome post-entry blocks, and assess pharmacologic agents to inhibit proteosome-mediated degradation of AAV.

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National Heart, Lung, and Blood Institute (NHLBI)
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Weill Medical College of Cornell University
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Ryan, Dorothy M; Vincent, Thomas L; Salit, Jacqueline et al. (2014) Smoking dysregulates the human airway basal cell transcriptome at COPD risk locus 19q13.2. PLoS One 9:e88051
Dvorak, Anna; Tilley, Ann E; Shaykhiev, Renat et al. (2011) Do airway epithelium air-liquid cultures represent the in vivo airway epithelium transcriptome? Am J Respir Cell Mol Biol 44:465-73
Krause, Anja; Whu, Wen Zhu; Xu, Yaqin et al. (2011) Protective anti-Pseudomonas aeruginosa humoral and cellular mucosal immunity by AdC7-mediated expression of the P. aeruginosa protein OprF. Vaccine 29:2131-9
Limberis, Maria P; Bell, Christie L; Heath, Jack et al. (2010) Activation of transgene-specific T cells following lentivirus-mediated gene delivery to mouse lung. Mol Ther 18:143-50
Limberis, M P; Bell, C L; Wilson, J M (2009) Identification of the murine firefly luciferase-specific CD8 T-cell epitopes. Gene Ther 16:441-7
Calcedo, Roberto; Vandenberghe, Luk H; Gao, Guangping et al. (2009) Worldwide epidemiology of neutralizing antibodies to adeno-associated viruses. J Infect Dis 199:381-90
Song, Yuhu; Lou, Howard H; Boyer, Julie L et al. (2009) Functional cystic fibrosis transmembrane conductance regulator expression in cystic fibrosis airway epithelial cells by AAV6.2-mediated segmental trans-splicing. Hum Gene Ther 20:267-81
Vandenberghe, L H; Breous, E; Nam, H-J et al. (2009) Naturally occurring singleton residues in AAV capsid impact vector performance and illustrate structural constraints. Gene Ther 16:1416-28
Fein, David E; Limberis, Maria P; Maloney, Sean F et al. (2009) Cationic lipid formulations alter the in vivo tropism of AAV2/9 vector in lung. Mol Ther 17:2078-87
Limberis, Maria P; Vandenberghe, Luk H; Zhang, Liqun et al. (2009) Transduction efficiencies of novel AAV vectors in mouse airway epithelium in vivo and human ciliated airway epithelium in vitro. Mol Ther 17:294-301

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