Single-dose lentiviral gene transfer for lifetime airway gene expression.
BACKGROUND
Cystic fibrosis (CF) is caused by a defect in cystic fibrosis transmembrane conductance regulator (CFTR) activity, often resulting in an incurable airway disease. Gene therapy into the conducting airway epithelium is a potential cure for CF; however, most gene vectors do not result in long-lived expression, and require re-dosing. Perversely, intrinsic host immune responses can then block renewed gene transfer.


METHODS
To investigate whether persistent gene expression could be achieved after a single dosing event, thus avoiding the issue of blocking host responses, we used a gene transfer protocol that combined an airway pretreatment using lysophosphatidylcholine with a human immunodeficiency virus type-1 (vesicular stomatitis virus G pseudotype) derived lentiviral vector to test whether an integrating vector could produce gene expression able to last for a substantial part of the lifetime of the laboratory mouse.


RESULTS
We found that a single dose of LV-LacZ produced immediate as well as lifetime mouse airway expression, confirming our hypothesis that use of an integrating vector extends transgene expression. Importantly, LV-CFTR dosing achieved at least 12 months of CFTR expression, representing partial functional correction of the CFTR defect in CF-null mice.


CONCLUSIONS
These findings validate the potential of this methodology for developing a gene transfer treatment for CF airway disease.
Stocker
Alice G
AG
Department of Respiratory and Sleep Medicine, Women's & Children's Hospital, Children, Youth & Women's Health Service, North Adelaide 5006, Australia.
Kremer
Karlea L
KL
Koldej
Rachel
R
Miller
Darren S
DS
Anson
Donald S
DS
Parsons
David W
DW
D000818:Animals; D002460:Cell Line; D003550:Cystic Fibrosis / Q000235:genetics / Q000378:metabolism / Q000628:therapy*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1 / Q000235:genetics; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008244:Lysophosphatidylcholines / Q000008:administration & dosage; D051379:Mice; D008810:Mice, Inbred C57BL; D018181:Mice, Inbred CFTR; D020545:Respiratory Mucosa / Q000187:drug effects / Q000378:metabolism*; D013997:Time Factors
D008244:Lysophosphatidylcholines; D019005:Cystic Fibrosis Transmembrane Conductance Regulator