Fibroblast growth factor 2 retargeted adenovirus has redirected cellular tropism: evidence for reduced toxicity and enhanced antitumor activity in mice.
Adenovirus (Ad) have been used as vectors to deliver genes to a wide variety of tissues. Despite achieving high expression levels in vivo, Ad vectors display normal tissue toxicity, transient expression, and antivector immune responses that limit therapeutic potential. To circumvent these problems, several retargeting strategies to abrogate native tropism and redirect Ad uptake through defined receptors have been attempted. Despite success in cell culture, in vivo results have generally not shown sufficient selectivity for target tissues. We have previously identified (C. K. Goldman et al., Cancer Res., 57: 1447-1451, 1997) the fibroblast growth factor (FGF) ligand and receptor families as conferring sufficient specificity and binding affinity to be useful for targeting DNA in vivo. In the present studies, we retargeted Ad using basic FGF (FGF2) as a targeting ligand. Cellular uptake is redirected through high-affinity FGF receptors (FGFRs) and not the more ubiquitous lower-affinity Ad receptors. Initial in vitro experiments demonstrated a 10- to 100-fold increase in gene expression in numerous FGFR positive (FGFR+) cell lines using FGF2-Ad when compared with Ad. To determine whether increased selectivity could be detected in vivo, FGF2-Ad was administered i.v. to normal mice. FGF2-Ad demonstrates markedly decreased hepatic toxicity and liver transgene expression compared with Ad treatment. Importantly, FGF2-Ad encoding the herpes simplex virus thymidine kinase (TK) gene transduces Ad-resistant FGFR+ tumor cells both ex vivo and in vivo, which results in substantially enhanced survival (180-260%) when the prodrug ganciclovir is administered. Because FGFRs are up-regulated on many types of malignant or injured cells, this broadly useful method to redirect native Ad tropism and to increase the potency of gene expression may offer significant therapeutic advantages.
Gu
D L
DL
Selective Genetics, Inc., San Diego, California 92121, USA. dlgu@selectivegenetics.com
Gonzalez
A M
AM
Printz
M A
MA
Doukas
J
J
Ying
W
W
D'Andrea
M
M
Hoganson
D K
DK
Curiel
D T
DT
Douglas
J T
JT
Sosnowski
B A
BA
Baird
A
A
Aukerman
S L
SL
Pierce
G F
GF
D000256:Adenoviridae / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D000410:Alanine Transaminase / Q000097:blood; D000818:Animals; D000998:Antiviral Agents / Q000627:therapeutic use; D001219:Aspartate Aminotransferases / Q000097:blood; D015415:Biomarkers / Q000097:blood; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D003673:Defective Viruses / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000008:administration & dosage / Q000276:immunology / Q000378:metabolism*; D015774:Ganciclovir / Q000627:therapeutic use; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000276:immunology / Q000627:therapeutic use*; D007140:Immunoglobulin Fab Fragments / Q000378:metabolism*; D000922:Immunotoxins / Q000008:administration & dosage / Q000378:metabolism* / Q000633:toxicity; D008099:Liver / Q000378:metabolism / Q000473:pathology / Q000821:virology; D008546:Melanoma, Experimental / Q000378:metabolism / Q000821:virology; D051379:Mice; D009336:Necrosis; D009928:Organ Specificity; D011991:Receptors, Virus / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000378:metabolism; D018139:Simplexvirus / Q000201:enzymology; D013937:Thymidine Kinase / Q000627:therapeutic use; D014162:Transfection; D019076:Transgenes; D018523:Tropism; D001616:beta-Galactosidase / Q000008:administration & dosage / Q000378:metabolism* / Q000633:toxicity
D000998:Antiviral Agents; D015415:Biomarkers; C568772:CLMP protein, mouse; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D007140:Immunoglobulin Fab Fragments; D000922:Immunotoxins; D011991:Receptors, Virus; D011993:Recombinant Fusion Proteins; D016222:Fibroblast Growth Factor 2; D001219:Aspartate Aminotransferases; D000410:Alanine Transaminase; D013937:Thymidine Kinase; D001616:beta-Galactosidase; D015774:Ganciclovir