Selective in vivo transfection of murine biliary epithelia using polycation-enhanced adenovirus.
We have investigated the use of polycations to increase adenovirus-mediated expression of transgenic protein to the biliary epithelia with a view to gene therapy for hepatobiliary disease in CF. We have shown that adenovirus carrying the beta-galactosidase transgene transfect both human and mouse biliary epithelia in primary culture and that in both instances adenovirus transfection can be significantly increased by co-complexing with polycation. In vivo administration of 1 x 109 p.f.u. adenovirus co-complexed with the polyamine polyethyenimine (PEI) into the mouse biliary duct leads to >80% positively stained biliary epithelia while adenovirus alone at the same titre infected <5% biliary epithelia. We suggest that the use of low titre polycation enhanced adenoviral delivery to the biliary tree of CF patients could be of therapeutic significance. As a prelude to an extensive in vivo functional investigation in CF null mice we have shown that Ad5/polycation complexes deliver functional CFTR to non-CFTR expressing cells in vitro more efficiently than Ad5 alone.
McKay
T R
TR
Cystic Fibrosis Gene Therapy Research Group, Molecular Genetics, Division of Biomedical Sciences, Imperial College School of Medicine, London, UK.
MacVinish
L J
LJ
Carpenter
B
B
Themis
M
M
Jezzard
S
S
Goldin
R
R
Pavirani
A
A
Hickman
M E
ME
Cuthbert
A W
AW
Coutelle
C
C
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002412:Cations; D002460:Cell Line; D046508:Culture Techniques; D003550:Cystic Fibrosis / Q000628:therapy*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D004848:Epithelium; D005704:Gallbladder; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage*; D006801:Humans; D051379:Mice; D014162:Transfection / Q000379:methods*
C505032:CFTR protein, human; D002412:Cations; D019005:Cystic Fibrosis Transmembrane Conductance Regulator