Epidermal growth factor targeting of bacteriophage to the choroid plexus for gene delivery to the central nervous system via cerebrospinal fluid.
Because the choroid plexus normally controls the production and composition of cerebrospinal fluid and, as such, its many functions of the central nervous system, we investigated whether ligand-mediated targeting could deliver genes to its secretory epithelium. We show here that when bacteriophages are targeted with epidermal growth factor, they acquire the ability to enter choroid epithelial cells grown in vitro as cell cultures, ex vivo as tissue explants or in vivo by intracerebroventricular injection. The binding and internalization of these particles activate EGF receptors on targeted cells, and the dose- and time-dependent internalization of particles is inhibited by the presence of excess ligand. When the phage genome is further reengineered to contain like green fluorescent protein or firefly luciferase under control of the cytomegalovirus promoter, gene expression is detectable in the choroid plexus and ependymal epithelium by immunohistochemistry or by noninvasive imaging, respectively. Taken together, these data support the hypothesis that reengineered ligand-mediated gene delivery should be considered a viable strategy to increase the specificity of gene delivery to the central nervous system and bypass the blood-brain barrier so as to exploit the biological effectiveness of the choroid plexus as a portal of entry into the brain.
Gonzalez
Ana Maria
AM
School of Clinical and Experimental Medicine, University of Birmingham, Edgbaston, UK.
Leadbeater
Wendy
W
Podvin
Sonia
S
Borboa
Alexandra
A
Burg
Michael
M
Sawada
Ritsuko
R
Rayner
James
J
Sims
Karen
K
Terasaki
Tetsuya
T
Johanson
Conrad
C
Stopa
Edward
E
Eliceiri
Brian
B
Baird
Andrew
A
D000818:Animals; D001435:Bacteriophages / Q000235:genetics; D002555:Cerebrospinal Fluid / Q000821:virology*; D002831:Choroid Plexus / Q000821:virology*; D004815:Epidermal Growth Factor / Q000235:genetics*; D004847:Epithelial Cells / Q000821:virology; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D007150:Immunohistochemistry; D051379:Mice; D051381:Rats; D014161:Transduction, Genetic
D004815:Epidermal Growth Factor