Lysophosphatidylcholine as an adjuvant for lentiviral vector mediated gene transfer to airway epithelium: effect of acyl chain length.
BACKGROUND
Poor gene transfer efficiency has been a major problem in developing an effective gene therapy for cystic fibrosis (CF) airway disease. Lysophosphatidylcholine (LPC), a natural airway surfactant, can enhance viral gene transfer in animal models. We examined the electrophysiological and physical effect of airway pre-treatment with variants of LPC on lentiviral (LV) vector gene transfer efficiency in murine nasal airways in vivo.


METHODS
Gene transfer was assessed after 1 week following nasal instillations of a VSV-G pseudotype LV vector pre-treated with a low and high dose of LPC variants. The electrophysiological effects of a range of LPC variants were assessed by nasal transepithelial potential difference measurements (TPD) to determine tight junction permeability. Any physical changes to the epithelium from administration of the LPC variants were noted by histological methods in airway tissue harvested after 1 hour.


RESULTS
Gene transduction was significantly greater compared to control (PBS) for our standard LPC (palmitoyl/stearoyl mixture) treatment and for the majority of the other LPC variants with longer acyl chain lengths. The LPC variant heptadecanoyl also produced significantly greater LV gene transfer compared to our standard LPC mixture. LV gene transfer and the transepithelial depolarization produced by the 0.1% LPC variants at 1 hour were strongly correlated (r2 = 0.94), but at the 1% concentration the correlation was less strong (r2 = 0.59). LPC variants that displayed minor to moderate levels of disruption to the airway epithelium were clearly associated with higher LV gene transfer.


CONCLUSIONS
These findings show the LPC variants effect on airway barrier function and their correlation to the effectiveness of gene expression. The enhanced expression produced by a number of LPC variants should provide new options for preclinical development of efficient airway gene transfer techniques.
Cmielewski
Patricia
P
Dept of Respiratory and Sleep Medicine, Women's and Children's Hospital, 72 King William Rd, North Adelaide, SA 5006, Australia. patricia.cmielewski@health.sa.gov.au
Anson
Don S
DS
Parsons
David W
DW
D000281:Administration, Intranasal; D000818:Animals; D005260:Female; D005822:Genetic Vectors; D007763:Lac Operon; D016086:Lentivirus / Q000235:genetics*; D008244:Lysophosphatidylcholines / Q000008:administration & dosage / Q000737:chemistry / Q000494:pharmacology*; D008564:Membrane Potentials; D051379:Mice; D008810:Mice, Inbred C57BL; D015394:Molecular Structure; D009297:Nasal Mucosa / Q000187:drug effects* / Q000378:metabolism; D010539:Permeability; D013501:Surface-Active Agents / Q000008:administration & dosage / Q000737:chemistry / Q000494:pharmacology*; D019108:Tight Junctions / Q000187:drug effects / Q000378:metabolism; D013997:Time Factors; D014161:Transduction, Genetic / Q000379:methods*
D008244:Lysophosphatidylcholines; D013501:Surface-Active Agents