Beta cyclodextrins enhance adenoviral-mediated gene delivery to the intestine.
OBJECTIVE
In general, the intestinal epithelium is quite refractory to viral and non-viral methods of gene transfer. In this report, various cyclodextrin formulations were tested for their ability to enhance adenoviral transduction efficiency in two models of the intestinal epithelium: differentiated Caco-2 cells and rat jejunum.


METHODS
Transduction efficiency of replication-deficient adenovirus type 5 vectors encoded with either the E. coli beta-galactosidase or the jellyfish green fluorescent protein gene was assessed by X-gal staining or visualization of fluorescence 48 hours after infection. In vivo experiments were performed using an intestinal loop ligation technique.


RESULTS
Several formulations of neutral and positively charged beta cyclodextrins significantly enhanced adenoviral-mediated gene transfer in the selected models. The cyclodextrin formulations studied increased adenoviral transduction in the intestine by enhancing both viral binding and internalization. Viral binding was significantly increased on cell membranes treated with positively charged cyclodextrins, as seen with confocal microscopy and rhodamine-labeled virus. Permeability studies and TEER readings revealed that the most successful formulations gently disrupt cell membranes. This enhances internalization of viral particles and results in increased levels of gene expression.


CONCLUSIONS
These formulations can be of value in gene transfer to cells and tissues in which adenoviral infection is limited due to a lack of fiber and alpha(v) integrin receptors. They are simple to prepare and do not affect the ability of the virus to transduce target cells.
College of Pharmacy, The University of Michigan, Ann Arbor 48109-1065, USA. macroyle@mail.med.upenn.edu
M A
MA
Croyle
B J
BJ
Roessler
C P
CP
Hsu
R
R
Sun
G L
GL
Amidon
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D018938:Caco-2 Cells; D002240:Carbohydrate Sequence; D002454:Cell Differentiation; D003505:Cyclodextrins / Q000494:pharmacology*; D017097:Electric Impedance; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D007413:Intestinal Mucosa / Q000187:drug effects / Q000378:metabolism*; D007583:Jejunum / Q000187:drug effects / Q000378:metabolism; D008969:Molecular Sequence Data; D009994:Osmolar Concentration; D051381:Rats; D017207:Rats, Sprague-Dawley
D003505:Cyclodextrins