title: Transduction of ovarian cancer cells: a recombinant adeno-associated viral vector compared to an adenoviral vector.
abstract: Recombinant adeno-associated virus (rAAV) vectors have emerged as vehicles for gene therapy. In addition, anti-neoplastic properties have been attributed to wild-type AAV. To take advantage of both features and to overcome technical problems associated with rAAV preparation, we developed a production method in which rAAV particles are amplified in an infectious cycle in the presence of wtAAV. This results in a 10(3)-10(4)-fold amplification of rAAV input particles. rAAV-GFP particles generated by this method were used to transduce ovarian cancer cell lines to evaluate their potential in ovarian cancer gene therapy, in comparison to a rAd-GFP vector. The transduction efficiency of NIH-OVCAR3, MDAH 2774 and SKOV3 cells with rAAV-GFP particles was low (< 1%) and did not improve by increasing the number of particles/cell. Repeated administration and continued exposure of NIH-OVCAR3 and MDAH 2774 improved transduction to over 3%. In contrast, these cell lines were more efficiently transduced by rAAV-GFP in the presence of adenovirus (approximately 15%) and by rAd-GFP (> 50%). These results indicate that in contrast to rAd vectors, rAAV particles are not suitable for therapeutic gene transfer in ovarian cancer cells unless efficient help can be provided to mediate ss to ds DNA conversion.
authors.affiliation: Laboratory of Medical and Molecular Oncology and Oncology Center, Akademisch Ziekenhuis Vrije Universiteit, Brussel, Belgium.
authors.forename: J
authors.initials: J
authors.lastname: Vermeij
authors.forename: Z
authors.initials: Z
authors.lastname: Zeinoun
authors.forename: B
authors.initials: B
authors.lastname: Neyns
authors.forename: E
authors.initials: E
authors.lastname: Teugels
authors.forename: C
authors.initials: C
authors.lastname: Bourgain
authors.forename: J
authors.initials: J
authors.lastname: De Grève
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D002277:Carcinoma / Q000378:metabolism* / Q000628:therapy; D004274:DNA, Recombinant / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D000229:Dependovirus / Q000235:genetics*; D005260:Female; D015316:Genetic Therapy; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics / Q000276:immunology; D021141:Nucleic Acid Amplification Techniques; D010051:Ovarian Neoplasms / Q000378:metabolism* / Q000628:therapy; D014161:Transduction, Genetic / Q000379:methods*; D014407:Tumor Cells, Cultured
chemical_list: D004274:DNA, Recombinant; D004279:DNA, Viral; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins