Optimizing aerosol gene delivery and expression in the ovine lung.
We have developed the sheep as a large animal model for optimizing cystic fibrosis gene therapy protocols. We administered aerosolized gene transfer agents (GTAs) to the ovine lung in order to test the delivery, efficacy, and safety of GTAs using a clinically relevant nebulizer. A preliminary study demonstrated GTA distribution and reporter gene expression throughout the lung after aerosol administration of plasmid DNA (pDNA):GL67 and pDNA:PEI complexes. A more comprehensive study examined the dose-response relationship for pDNA:PEI and assessed the influence of adjunct therapeutic agents. We found that the sheep model can differentiate between doses of GTA and that the anticholinergic, glycopyrrolate, enhanced transgene expression. Dose-related toxicity of GTA was reduced by aerosol administration compared to direct instillation. This large animal model will allow us to move toward clinical studies with greater confidence.
McLachlan
Gerry
G
Medical Genetics Section, School of Molecular and Clinical Medicine, University of Edinburgh, Western General Hospital, Edinburgh, UK. gerry.mclachlan@ed.ac.uk
Baker
Alison
A
Tennant
Peter
P
Gordon
Catherine
C
Vrettou
Christina
C
Renwick
Louise
L
Blundell
Richard
R
Cheng
Seng H
SH
Scheule
Ron K
RK
Davies
Lee
L
Painter
Hazel
H
Coles
Rebecca L
RL
Lawton
Anna E
AE
Marriott
Chris
C
Gill
Deborah R
DR
Hyde
Stephen C
SC
Griesenbach
Uta
U
Alton
Eric W F W
EW
Boyd
A Christopher
AC
Porteous
David J
DJ
Collie
D David S
DD
D000818:Animals; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D018014:Gene Transfer Techniques / Q000009:adverse effects / Q000295:instrumentation*; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D008168:Lung / Q000378:metabolism*; D008171:Lung Diseases / Q000209:etiology / Q000473:pathology; D008297:Male; D009330:Nebulizers and Vaporizers; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012756:Sheep
D011993:Recombinant Fusion Proteins; D015500:Chloramphenicol O-Acetyltransferase