Signaling antibodies complexed with adenovirus circumvent CAR and integrin interactions and improve gene delivery.
Current adenoviral (Ad) vectors cannot be targeted to specific cell types due to the widespread distribution of the Ad receptor (CAR). Moreover, CAR and/or internalization receptors (alphav integrins) are absent or present at low levels on some cell types, rendering them resistant to Ad-mediated gene delivery. To address these problems, we have developed a novel vector targeting approach that takes advantage of the common cell signaling pathways initiated by ligation of alphav integrins and growth factor receptors. Recombinant growth factor/cytokines (TNF-alpha, IGF-1, EGF) which trigger phosphatidylinositol-3-OH kinase (PI3K) activation, a signaling molecule involved in adenovirus internalization, were fused to a monoclonal antibody specific for the viral penton base. Ad vectors complexed with these bifunctional mAbs increased gene delivery 10 to 50-fold to human melanoma cells lacking alphav integrins. The bifunctional mAbs also enhanced gene delivery by fiberless adenovirus particles which cannot bind to CAR. Improved gene delivery correlated with increased virus internalization and attachment as well as PI3K activity. The use of bifunctional mAbs to trigger specific cell signaling pathways offers a widely applicable method for bypassing the normal Ad receptors in gene delivery and potentially increasing the selectivity of gene transfer.
Li
E
E
Department of Immunology, The Scripps Research Institute, La Jolla, CA 92037, USA.
Brown
S L
SL
Von Seggern
D J
DJ
Brown
G B
GB
Nemerow
G R
GR
D000256:Adenoviridae / Q000235:genetics* / Q000276:immunology; D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D015179:Colorectal Neoplasms / Q000378:metabolism / Q000628:therapy; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004815:Epidermal Growth Factor; D015870:Gene Expression; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage*; D006801:Humans; D007313:Insecta; D007334:Insulin-Like Growth Factor I; D039564:Integrin alphaV; D008545:Melanoma / Q000378:metabolism / Q000628:therapy; D009369:Neoplasms / Q000201:enzymology / Q000628:therapy*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011991:Receptors, Virus / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage; D015398:Signal Transduction / Q000276:immunology*; D014162:Transfection / Q000379:methods; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology; D000077191:Wortmannin; D001616:beta-Galactosidase / Q000235:genetics
D000730:Androstadienes; D015703:Antigens, CD; D004791:Enzyme Inhibitors; D039564:Integrin alphaV; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011991:Receptors, Virus; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; C105941:coxsackievirus B receptor; D004815:Epidermal Growth Factor; D007334:Insulin-Like Growth Factor I; D001616:beta-Galactosidase; D000077191:Wortmannin