title: Evaluation and optimization of the administration of recombinant adeno-associated viral vectors (serotypes 2/1, 2/2, 2/rh8, 2/9, and 2/rh10) by convection-enhanced delivery to the striatum.
abstract: Convection-enhanced delivery (CED) of recombinant adeno-associated virus (rAAV) vectors is a promising approach for delivery of therapeutic transgenes to the brain. In this study we have systematically examined vector dosing in vivo. Infusions of rAAV serotypes 2/1, 2/2, 2/rh8, 2/9, and 2/rh10 expressing an enhanced green fluorescent protein reporter gene were undertaken into the striatum of rats and pigs using CED. Vector distribution, as defined by the volume of distribution and number of transduced cells following each infusion, was determined using stereological methods. Immunohistochemistry was used to determine the transductional tropism of serotypes and to evaluate for the presence of immune cell infiltration into the brain. Vector distribution was highly variable between serotypes. Infusion rate had no significant effect on vector distribution or the occurrence of tissue damage. For serotypes 2/1, 2/2 and 2/rh10, as the vector concentration was increased beyond 10(12) vg/ml, no increase in vector distribution was observed. In contrast, for serotypes 2/rh8 and 2/9, retrograde axonal transport was observed above this threshold concentration. Cell transduction was principally neuronal for all serotypes and was associated with a low-level immune response. In planning clinical trials it is critical that these observations are considered in order to achieve optimal vector dosing.
authors.affiliation: Department of Neurosurgery, Frenchay Hospital, Bristol, UK.
authors.forename: Edward
authors.initials: E
authors.lastname: White
authors.forename: Alison
authors.initials: A
authors.lastname: Bienemann
authors.forename: Miguel
authors.initials: M
authors.lastname: Sena-Esteves
authors.forename: Hannah
authors.initials: H
authors.lastname: Taylor
authors.forename: Chotirote
authors.initials: C
authors.lastname: Bunnun
authors.forename: Emma
authors.initials: E
authors.lastname: Castrique
authors.forename: Steven
authors.initials: S
authors.lastname: Gill
mesh_terms: D000818:Animals; D018481:Convection; D003342:Corpus Striatum / Q000821:virology*; D000229:Dependovirus / Q000235:genetics* / Q000378:metabolism; D015316:Genetic Therapy; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D023421:Models, Animal; D009474:Neurons / Q000378:metabolism* / Q000821:virology*; D051381:Rats; D017208:Rats, Wistar; D013552:Swine; D014161:Transduction, Genetic; D019076:Transgenes
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins