Novel Immune Response Evasion Strategy to Redose Adeno-Associated Viral Vectors and Prolong Survival in Surfactant Protein-B Deficient Mice

The goals and objectives for this study are to determine whether AAV containing TLR9 inhibitory oligonucleotide sequences (AAV-hSPBTLR9i) can mitigate the immune response sufficiently to redose AAV in the lungs and prolong the survival of SP-B deficient mice. 24 samples were analyzed for 773 murine genes plus 12 internal reference genes. There were 6 treatment groups: (1) no treatment - on doxycycline (positive control), (2) no treatment - off doxycycline (negative control), (3) AAV-hSPB single, (4) AAV-hSPBTLR9i single, (5) AAV-hSPB repeat, (6) AAV-hSPBTLR9i repeat. Groups 3-6 were administered their corresponding treatment via intratracheal delivery on Day 0. Doxycyline feed was removed from groups 3-6 on Day 7. Ninety days after the first AAV dose, groups 5 & 6 were given their second AAV dose via intratracheal delivery. Doxycycline feed was removed from group 2 on Day 94. On Day 97, the lungs were harvested from the mice and flash frozen. Then RNA was extracted from the lungs for gene expression analysis and interferon gene expression (nCounter). 9 samples were analyzed for 773 murine genes plus 12 internal reference genes. There were 3 treatment groups: (1) 1xPBS (positive control), (2) AAV-hSPB single, (3) AAV-hSPBTLR9i single. Mice were given their corresponding AAV treatment via intratracheal delivery on Day 0. Three hours later, the lungs of the mice were harvested and flash frozen. RNA was extracted from the lungs for gene expression analysis (nCounter).