title: Recombinant adeno-associated virus: efficient transduction of the rat VMH and clearance from blood.
abstract: To promote the efficient and safe application of adeno-associated virus (AAV) vectors as a gene transfer tool in the central nervous system (CNS), transduction efficiency and clearance were studied for serotypes commonly used to transfect distinct areas of the brain. As AAV2 was shown to transduce only small volumes in several brain regions, this study compares the transduction efficiency of three AAV pseudotyped vectors, namely AAV2/1, AAV2/5 and AAV2/8, in the ventromedial nucleus of the hypothalamus (VMH). No difference was found between AAV2/1 and AAV2/5 in transduction efficiency. Both AAV2/1 and AAV2/5 achieved a higher transduction rate than AAV2/8. One hour after virus administration to the brain, no viral particles could be traced in blood, indicating that no or negligible numbers of virions crossed the blood-brain barrier. In order to investigate survival of AAV in blood, clearance was determined following systemic AAV administration. The half-life of AAV2/1, AAV2/2, AAV2/5 and AAV2/8 was calculated by determining virus clearance rates from blood after systemic injection. The half-life of AAV2/2 was 4.2 minutes, which was significantly lower than the half-lives of AAV2/1, AAV2/5 and AAV2/8. With a half-life of more than 11 hours, AAV2/8 particles remained detectable in blood significantly longer than AAV2/5. We conclude that application of AAV in the CNS is relatively safe as no AAV particles are detectable in blood after injection into the brain. With a half-life of 1.67 hours of AAV2/5, a systemic injection with 1×109 genomic copies of AAV would be fully cleared from blood after 2 days.
authors.affiliation: Brain Center Rudolf Magnus, Department of Translational Neuroscience, University Medical Center Utrecht, University of Utrecht, Utrecht, The Netherlands.
authors.forename: Margriet A
authors.initials: MA
authors.lastname: van Gestel
authors.affiliation: Brain Center Rudolf Magnus, Department of Translational Neuroscience, University Medical Center Utrecht, University of Utrecht, Utrecht, The Netherlands.
authors.forename: Arjen J
authors.initials: AJ
authors.lastname: Boender
authors.affiliation: Brain Center Rudolf Magnus, Department of Translational Neuroscience, University Medical Center Utrecht, University of Utrecht, Utrecht, The Netherlands.
authors.forename: Veronne A J
authors.initials: VA
authors.lastname: de Vrind
authors.affiliation: Brain Center Rudolf Magnus, Department of Translational Neuroscience, University Medical Center Utrecht, University of Utrecht, Utrecht, The Netherlands.
authors.forename: Keith M
authors.initials: KM
authors.lastname: Garner
authors.affiliation: Brain Center Rudolf Magnus, Department of Translational Neuroscience, University Medical Center Utrecht, University of Utrecht, Utrecht, The Netherlands.
authors.forename: Mieneke C M
authors.initials: MC
authors.lastname: Luijendijk
authors.affiliation: Brain Center Rudolf Magnus, Department of Translational Neuroscience, University Medical Center Utrecht, University of Utrecht, Utrecht, The Netherlands.
authors.forename: Roger A H
authors.initials: RA
authors.lastname: Adan
mesh_terms: D000818:Animals; D001769:Blood / Q000821:virology*; D004274:DNA, Recombinant / Q000235:genetics*; D000229:Dependovirus / Q000235:genetics* / Q000502:physiology; D057809:HEK293 Cells; D006801:Humans; D051381:Rats; D014018:Tissue Distribution; D014161:Transduction, Genetic; D014697:Ventromedial Hypothalamic Nucleus / Q000821:virology*
chemical_list: D004274:DNA, Recombinant