Lentiviral airway gene transfer in lungs of mice and sheep: successes and challenges.
BACKGROUND
Persistent airway gene expression can be achieved in mouse nasal airway using a vesicular stomatitis virus glycoprotein pseudotyped lentiviral (LV) gene vector in combination with lysophosphatidylcholine (LPC) pretreatment. We have now examined the acute in vivo effects of this combination single-dose method for airway LV gene transfer in mouse and sheep lung.


METHODS
Mouse and sheep lungs were exposed to LPC followed 1 h later with the LV vector. Lungs were processed 7 days later using X-gal detection to measure beta-gal gene expression and identify transduced cell types.


RESULTS
In mouse ciliated conducting airways, LPC pretreatment produced extensive gene transfer that extended from the tracheal dosing site into the bronchi and lower airways. Gene expression was present in both terminally differentiated surface cells and in basal cells. Without LPC pretreatment, transduction was limited to the dosing site. In sheep lung, small-volume bronchoscopic instillation delivery produced localized and low-level transduction near the dosing site. Gene expression was again present in surface and basal cells. Neither alterations in LPC dose parameters, nor larger vector volumes increased the level of transduction.


CONCLUSIONS
These findings are the first to confirm the applicability of LPC pretreatment in the production of extensive lentiviral gene transfer in mouse lung airways. However, improved methodologies to increase transduction efficiency are required for adult sheep lung. The results suggest that continued in vivo development of LPC-enhanced lentiviral gene transfer is needed in the lungs of large animals to establish effective lentiviral gene transfer techniques suited to the treatment of airway disease.
Liu
Chuanhe
C
Department of Respiratory and Sleep Medicine, Women's and Children's Hospital, Adelaide, Australia.
Wong
Edward
E
Miller
Darren
D
Smith
Greg
G
Anson
Don
D
Parsons
David
D
D000818:Animals; D005260:Female; D015870:Gene Expression; D015316:Genetic Therapy; D005822:Genetic Vectors; D016086:Lentivirus / Q000235:genetics*; D008168:Lung / Q000378:metabolism*; D008244:Lysophosphatidylcholines / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D012137:Respiratory System / Q000378:metabolism; D012756:Sheep; D014161:Transduction, Genetic
D008244:Lysophosphatidylcholines