Gene delivery to airway epithelial cells in vivo: a direct comparison of apical and basolateral transduction strategies using pseudotyped lentivirus vectors.
Lentivirus vectors are being investigated as gene delivery vehicles for cystic fibrosis airway gene therapy. Vesicular stomatitis virus G glycoprotein (VSV-G)-pseudotyped vectors transduce airway epithelia via receptors that are located predominantly on the basolateral surface of the airway epithelium. Effective transduction with VSV-G-pseudotyped vectors requires the use of a pre-treatment that disrupts epithelial tight junctions, allowing access to these basolateral receptors. In contrast, it has been reported that apically targeted lentiviral vectors allow efficient gene transfer in the absence of any pre-treatment. In a direct comparison of transduction by a VSV-G-pseudotyped vector, in combination with a pre-treatment with lysophosphatidylcholine (LPC), and the same vector pseudotyped with the apically targeted baculovirus GP64 envelope (without any pre-treatment), the GP64 vector was found to be significantly less efficient. However, when a pre-treatment with LPC was used the level of transduction with the GP64-pseudotyped lentiviral vector was not significantly different to that resulting from the VSV-G-pseudotyped vector. The cell types transduced with each vector were essentially the same, with the majority of cells transduced being respiratory (ciliated cells). However, unlike the VSV-G-pseudotyped vector, which results in persisting gene expression, transduction with the GP64-pseudotyped vector resulted in gene expression that declined to undetectable levels over six months, whether or not an LPC pre-treatment was used.
Kremer
Karlea L
KL
Department of Genetic Medicine, Women's and Children's Hospital, CYWHS, 72 King William Road, North Adelaide, South Australia.
Dunning
Kylie R
KR
Parsons
David W
DW
Anson
Donald S
DS
D015815:Cell Adhesion Molecules / Q000235:genetics; D016764:Cell Polarity; D003550:Cystic Fibrosis / Q000628:therapy; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008244:Lysophosphatidylcholines / Q000494:pharmacology; D008562:Membrane Glycoproteins / Q000235:genetics; D012137:Respiratory System / Q000166:cytology; D019108:Tight Junctions / Q000187:drug effects; D014161:Transduction, Genetic / Q000379:methods* / Q000592:standards; D014759:Viral Envelope Proteins / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics
D015815:Cell Adhesion Molecules; C028391:G protein, vesicular stomatitis virus; D008244:Lysophosphatidylcholines; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins; D014764:Viral Proteins