Vascular bed-targeted in vivo gene delivery using tropism-modified adeno-associated viruses.
Virus-mediated gene delivery is restricted by the infectivity profile of the chosen vector. Targeting the vascular endothelium via systemic delivery has been attempted using peptides isolated in vitro (using either phage or vector display) and implicit reliance on target receptor expression in vivo. This has limited application since endothelial cells in vitro and in vivo differ vastly in receptor profiles and because of the existence of complex endothelial "zip codes" in vivo. We therefore tested whether in vivo phage display combined with adeno-associated virus (AAV) capsid modifications would allow in vivo homing to the endothelium residing in defined organs. Extensive in vivo biopanning in rats identified four consensus peptides homing to the lung or brain. Each was incorporated into the VP3 region of the AAV-2 capsid to display the peptide at the virion surface. Peptides that conferred heparan independence were shown to retarget virus to the expected vascular bed in vivo in a preferential manner, determined 28 days post-systemic injection by both virion DNA and transgene expression profiling. Our findings significantly impact the design of viral vectors for targeting individual vascular beds in vivo.
Work
Lorraine M
LM
BHF Glasgow Cardiovascular Research Centre, Division of Cardiovascular and Medical Sciences, University of Glasgow, 126 University Avenue, Glasgow G12 8TA, UK.
Büning
Hildegard
H
Hunt
Ela
E
Nicklin
Stuart A
SA
Denby
Laura
L
Britton
Nicola
N
Leike
Kristen
K
Odenthal
Margarete
M
Drebber
Uta
U
Hallek
Michael
M
Baker
Andrew H
AH
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002213:Capsid / Q000737:chemistry / Q000378:metabolism; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016384:Consensus Sequence; D004279:DNA, Viral / Q000032:analysis; D000229:Dependovirus / Q000235:genetics*; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism* / Q000821:virology; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors; D006497:Heparan Sulfate / Q000378:metabolism; D007150:Immunohistochemistry; D007763:Lac Operon; D008099:Liver / Q000821:virology; D008297:Male; D016254:Mutagenesis, Insertional; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000235:genetics*; D017433:Protein Structure, Secondary; D051381:Rats; D011921:Rats, Inbred WKY; D017386:Sequence Homology, Amino Acid; D013997:Time Factors; D019076:Transgenes; D014771:Virion / Q000235:genetics / Q000378:metabolism
D036022:Capsid Proteins; D004279:DNA, Viral; D006497:Heparan Sulfate; D019151:Peptide Library; D010455:Peptides