title: scAAV-mediated gene transfer of interleukin-1-receptor antagonist to synovium and articular cartilage in large mammalian joints.
abstract: With the long-term goal of developing a gene-based treatment for osteoarthritis (OA), we performed studies to evaluate the equine joint as a model for adeno-associated virus (AAV)-mediated gene transfer to large, weight-bearing human joints. A self-complementary AAV2 vector containing the coding regions for human interleukin-1-receptor antagonist (hIL-1Ra) or green fluorescent protein was packaged in AAV capsid serotypes 1, 2, 5, 8 and 9. Following infection of human and equine synovial fibroblasts in culture, we found that both were only receptive to transduction with AAV1, 2 and 5. For these serotypes, however, transgene expression from the equine cells was consistently at least 10-fold higher. Analyses of AAV surface receptor molecules and intracellular trafficking of vector genomes implicate enhanced viral uptake by the equine cells. Following delivery of 1 × 10(11) vector genomes of serotypes 2, 5 and 8 into the forelimb joints of the horse, all three enabled hIL-1Ra expression at biologically relevant levels and effectively transduced the same cell types, primarily synovial fibroblasts and, to a lesser degree, chondrocytes in articular cartilage. These results provide optimism that AAV vectors can be effectively adapted for gene delivery to large human joints affected by OA.
authors.affiliation: Department of Orthopaedics and Rehabilitation, University of Florida, Gainesville, FL 32608-0137, USA.
authors.forename: R S
authors.initials: RS
authors.lastname: Watson
authors.forename: T A
authors.initials: TA
authors.lastname: Broome
authors.forename: P P
authors.initials: PP
authors.lastname: Levings
authors.forename: B L
authors.initials: BL
authors.lastname: Rice
authors.forename: J D
authors.initials: JD
authors.lastname: Kay
authors.forename: A D
authors.initials: AD
authors.lastname: Smith
authors.forename: E
authors.initials: E
authors.lastname: Gouze
authors.forename: J-N
authors.initials: JN
authors.lastname: Gouze
authors.forename: E A
authors.initials: EA
authors.lastname: Dacanay
authors.forename: W W
authors.initials: WW
authors.lastname: Hauswirth
authors.forename: D M
authors.initials: DM
authors.lastname: Nickerson
authors.forename: M J
authors.initials: MJ
authors.lastname: Dark
authors.forename: P T
authors.initials: PT
authors.lastname: Colahan
authors.forename: S C
authors.initials: SC
authors.lastname: Ghivizzani
mesh_terms: D000818:Animals; D002358:Cartilage, Articular / Q000378:metabolism / Q000473:pathology / Q000821:virology; D000229:Dependovirus / Q000235:genetics*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D006736:Horses; D006801:Humans; D053590:Interleukin 1 Receptor Antagonist Protein / Q000235:genetics*; D007375:Interleukin-1 / Q000235:genetics; D007596:Joints / Q000378:metabolism / Q000473:pathology / Q000821:virology; D010003:Osteoarthritis / Q000235:genetics* / Q000628:therapy; D013583:Synovial Membrane / Q000378:metabolism / Q000473:pathology / Q000821:virology
chemical_list: D053590:Interleukin 1 Receptor Antagonist Protein; D007375:Interleukin-1; D049452:Green Fluorescent Proteins