Abstract
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 β-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 × 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.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Park RW, Grand RJ . Gastrointestinal manifestations of cystic fibrosis: a review Gastroenterology 1981 81: 1143–1161
Vawter GF, Shwachman H . Cystic fibrosis in adults: an autopsy study Pathol Annu 1979 14: 357–382
Schuster SR et al. The management of portal hypertension in cystic fibrosis J Pediatr Surg 1977 12: 201–206
Grubman SA et al. Correction of the cystic fibrosis defect by gene complementation in human intrahepatic biliary epithelial cell lines Gastroenterology 1995 108: 584–592
Cohn JA et al. Localization of the cystic fibrosis transmembrane conductance regulator in human bile duct epithelial cells Gastroenterology 1993 105: 1857–1864
Yang Y et al. An approach for treating the hepatobiliary disease of cystic fibrosis by somatic gene transfer Proc Natl Acad Sci USA 1993 90: 4601–4605
Maeda H, Danel C, Crystal RG . Adenovirus-mediated transfer of human lipase complementary DNA to the gallbladder Gastroenterology 1994 106: 1638–1644
Dray-Charier N et al. Regulation of mucin secretion in human gallbladder epithelial cells: predominant role of calcium and protein kinase C Gastroenterology 1997 112: 978–990
Vickers SM et al. In vivo gene transfer to the human biliary tract Gene Therapy 1996 3: 825–828
Curtis CM et al. Restoration by intratracheal gene transfer of bicarbonate secretion in cystic fibrosis mouse gallbladder Am J Physiol 1998 274: G1053–G1060
Fasbender A et al. Complexes of adenovirus with polycationic polymers and cationic lipids increase the efficiency of gene transfer in vitro and in vivo J Biol Chem 1997 272: 6479–6489
Kaplan JM et al. Potentiation of gene transfer to the mouse lung by complexes of adenovirus vector and polycations improves therapeutic potential Hum Gene Ther 1998 9: 1469–1479
Vrancken Peeters MJ, Perkins AL, Kay MA . Method for multiple portal vein infusions in mice: quantitation of adenovirus-mediated hepatic gene transfer Biotechniques 1996 20: 278–285
Anderson MP et al. Demonstration that CFTR is a chloride channel by alteration of its anion selectivity Science 1991 253: 202–205
Bear CE et al. Purification and functional reconstitution of the cystic fibrosis transmembrane conductance regulator (CFTR) Cell 1992 68: 809–818
Seamon K, Daly JW . Activation of adenylate cyclase by the diterpene forskolin does not require the guanine nucleotide regulatory protein J Biol Chem 1981 256: 9799–9801
Kochanek S et al. Replacement of all viral coding sequences with 28 kb of DNA independently expressing both full-length dystrophin and beta-galactosidase Proc Natl Acad Sci USA 1996 93: 5731–5736
Feng M et al. Stable in vivo gene transduction via a novel adenoviral/retroviral chimeric vector Nat Biotechnol 1997 15: 866–870
MacVinish LJ et al. Kinin-induced chloride permeability changes in colony 29 epithelia estimated from 125I-efflux and MEQfluorescence Br J Pharmacol 1993 108: 469–478
Henderson RM, Ashford ML, MacVinish LJ, Cuthbert AW . Chloride channels and anion fluxes in a human colonic epithelium (HCA-7) Br J Pharmacol 1992 106: 109–114
Acknowledgements
T McKay et al would like to thank Anne-Marie Douar for her help early in this project and Professor Knuth for providing the Mz-ChA-2 cell line. This work has been funded by the CF-Trust and MRC.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
McKay, T., MacVinish, L., Carpenter, B. et al. Selective in vivo transfection of murine biliary epithelia using polycation-enhanced adenovirus. Gene Ther 7, 644–652 (2000). https://doi.org/10.1038/sj.gt.3301144
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.gt.3301144
