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. Copyright 2010 John Wiley & Sons, Ltd.
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. Copyright 2010 John Wiley & Sons, Ltd.
Authors: David Parsons; Martin Donnelley; Alexandra McCarron; Patricia Cmielewski; Victoria Drysdale Journal: Gene Ther Date: 2022-03-29 Impact factor: 4.184
Authors: Tessa Gargett; Branka Grubor-Bauk; Darren Miller; Tamsin Garrod; Stanley Yu; Steve Wesselingh; Andreas Suhrbier; Eric J Gowans Journal: Clin Transl Immunology Date: 2014-06-27