Adeno-associated viral vectors based on serotype 3b use components of the fibroblast growth factor receptor signaling complex for efficient transduction.
Adeno-associated virus type 3b (AAV3b) has been largely ignored by gene therapists because of the inability of vectors based on this serotype to transduce target tissues efficiently. Here we describe a phenomenon unique to AAV3b in that vectors based on this serotype mediate enhanced transduction in the presence of heparin. Among the many biological functions attributed to heparin, its interaction with, and ability to regulate, several growth factors (GFs) and growth factor receptors (GFRs) has been well characterized. Using GFR-overexpressing cell lines, soluble GFs and heparins, as well as specific GFR inhibitors, we have demonstrated a requirement for fibroblast growth factor receptor-2 (FGFR2) and FGF1 in the heparin-mediated augmentation of AAV3b vector transduction. In contrast to AAV2, we establish that heparin can be used as an adjunct with AAV3b to further increase transduction in a variety of cells and target tissues, additionally suggesting that AAV3b may be an attractive viral vector for clinical use during procedures in which heparin is used. In summary, AAV3b exhibits FGFR2-dependent, markedly enhanced transduction efficiency in the presence of heparin and FGFs, which could make it a useful vector for gene therapy in a variety of human diseases.
Cardiothoracic Division, Department of Surgery, Duke University Medical Center, Durham, NC 27710, USA.
Emily L
EL
Messina
Jeffrey
J
Nienaber
Mani
M
Daneshmand
Nestor
N
Villamizar
Jude
J
Samulski
Carmelo
C
Milano
Dawn E
DE
Bowles
D000818:Animals; D045744:Cell Line, Tumor; D000229:Dependovirus / Q000145:classification* / Q000187:drug effects / Q000235:genetics*; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics*; D005838:Genotype; D006025:Glycosaminoglycans / Q000378:metabolism; D006493:Heparin / Q000008:administration & dosage / Q000494:pharmacology; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D051497:Receptor, Fibroblast Growth Factor, Type 2 / Q000235:genetics / Q000378:metabolism*; D012501:Saphenous Vein / Q000187:drug effects / Q000378:metabolism; D012703:Serotyping; D012995:Solubility; D034421:Sus scrofa; D046509:Tissue Culture Techniques; D014161:Transduction, Genetic / Q000379:methods*
D006025:Glycosaminoglycans; D036341:Intercellular Signaling Peptides and Proteins; D006493:Heparin; D008156:Luciferases; D051497:Receptor, Fibroblast Growth Factor, Type 2