Molecular Therapy, Vol 18, S1, Abstract 873
The American Society of Gene and Cell Therapy Annual Conference, Washington DC, 2010
We have previously shown that simian immunodefciency virus pseudotyped with F and HN protein from Sendai virus (F/HN-SIV) transduces murine nasal epithelium effciently (3-5% of respiratory epithelial cells after perfusion with 5x108 TU) and importantly can be repeatedly administered.
We now show that expression in the nasal epithelium persists for the life-time of the animals (16-25 months, 9 of 9 C57Bl/6 mice). The vast majority of published studies have assessed lentiviral vectors in mouse nasal epithelium, and transduction of lung epithelium, particularly without pre-conditioning through polidocanol treatment or tissue damage, remains challenging. Here, we show that F/HN-SIV transduces lung epithelium effciently and dose-dependently.
In contrast to other studies, we show that lentivirus-mediated gene expression in the lung is stable for at least 13 months (latest time-point currently analyzed) (month 2: 362660±63922photons/sec, month 13: 431454±65647photons/sec, n=5-8 mice). We also show, for the frst time, that lentivirus can be repeatedly administered to the lung (2 doses of F/HN-SIV-GFP followed by a 3rd dose of F/HN-SIV-Lux at monthly interval) without loss of activity compared to a single dose of F/HN-SIV-Lux (1 Dose: 40178±4843, 3 Doses: 39080±7490 RLU/mg protein, n=21/ group).
Importantly, we also show that gene expression increases with increasing number of doses (10 doses at daily interval) (Dose 1:4693± 899, Dose 10: 239212±48362 RLU/mg protein, n=8/group, p< 0.0005). Importantly, we have not observed acute or chronic toxicity in 12 months follow-up studies in mice.
We have also assessed the performance of F/HN-SIV in various airway ex vivo models.
These data suggest that, F/HN-SIV may be a suitable vector for cystic fbrosis gene therapy.