Does the use of recombinant AAV5 in pulmonary gene therapy lead to lung damage?

• Published in: Respiratory physiology & neurobiology Vol. 168; no. 3; pp. 203 - 209
• Main Authors: Martini, S.V, Fagundes, S.S, Schmidt, A.C, Avila, M, Ornellas, D.S, Ribas, V.T, Petrs-Silva, H, Linden, R, Faffe, D.S, Guggino, S.E, Rocco, P.R.M, Zin, W.A, Morales, M.M
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 30.09.2009
• Subjects: Adeno-associated virus 5; Adeno-associated virus type 5; Airway Resistance; Analysis of Variance; Animals; Apoptosis; Disease Models, Animal; Gene therapy; Genetic Therapy - adverse effects; Genetic Vectors - adverse effects; Green Fluorescent Proteins - genetics; In Situ Nick-End Labeling; Male; Medical Education; Mice; Mice, Inbred BALB C; Pneumonia - etiology; Pneumonia - pathology; Pulmonary mechanics; Pulmonary/Respiratory; Respiratory Mechanics - physiology; RNA, Messenger - metabolism; Time Factors; Vector; polymorphonuclear cells; equipment resistive pressure; pulmonary viscoelastic/inhomogeneous pressure; complementary deoxyribonucleic acid; virus group; adeno-associated virus; control group; V T; Δ P1; Δ P2; GFP; recombinant adeno-associated virus type 5; rAAV5; cystic fibrosis; RT-PCR; V; pulmonary resistive pressure; cDNA; glyceraldehyde-3-phosphate dehydrogenase; tidal volume; Req· V; Vector; diethylpyro-carbonate-treated water; reverse-transcription polymerase chain reaction; AAV5; CF; Est; resistance of the equipment; airflow; DEPC; adeno-associated virus type 5; ITRs; PMN; pulmonary elastance; CTRL; AAV; inverted terminal repeats; cystic fibrosis transmembrane conductance regulator gene; green fluorescence protein; VR; Gene therapy; GAPDH; CFTR; Pulmonary mechanics; Req; Adeno-associated virus type 5
• ISSN: 1569-9048; 1878-1519
• DOI: 10.1016/j.resp.2009.06.016

Abstract This study investigated whether repeated administration of recombinant adeno-associated virus type 5 (rAAV5) to the airways induces inflammatory processes in the lungs of BALB/c-mice, with mechanical and histologic changes. Saline was instilled intratracheally in the control group, and rAAV5-green fluorescence protein (GFP) (4 × 1011 particles) in the virus group (VR). These groups were subdivided into four subgroups: one dose analyzed 3 weeks later (VR1d3w) and two doses analyzed 1 (VR2d1w), 2 (VR2d2w) and 3 weeks (VR2d3w) after the second dose. Lung morphometry, mechanical parameters, airway responsiveness, rAAV5-GFP transduction and the expression of inflammatory cytokines were investigated. No significant differences in lung mechanics, airway responsiveness, and morphometry were observed. Re-administration of rAAV5 vector resulted in a decrease in GFP mRNA expression in the VR2d3w group. There was no evidence of inflammatory response or apoptosis in any group. rAAV5 did not induce an inflammatory process, mechanical or morphometric changes in the lungs. AAV5 may be an appropriate vector for lung gene therapy.