Intra-articular gene delivery and expression of interleukin-1Ra mediated by self-complementary adeno-associated virus
Abbreviated Journal Title
J. Gene. Med.
adeno-associated virus; arthritis; gene therapy; interleukin-1; interleukin-1 receptor antagonist; COLLAGEN-INDUCED ARTHRITIS; TUMOR-NECROSIS-FACTOR; RATE-LIMITING STEP; RECEPTOR-ANTAGONIST; IN-VIVO; INTRACELLULAR TRAFFICKING; EFFICIENT; TRANSDUCTION; TRANSGENE EXPRESSION; RHEUMATOID-ARTHRITIS; MURINE; FIBROBLASTS; Biotechnology & Applied Microbiology; Genetics & Heredity; Medicine, ; Research & Experimental
Background The adeno-associated Virus (AAV) has many safety features that favor its use in the treatment of arthritic conditions; however, the conventional, single-stranded vector is inefficient for gene delivery to fibroblastic cells that primarily populate articular tissues. This has been attributed to the inability of these cells to convert the vector to a double-stranded form. To overcome this, we evaluated double-stranded self-complementary (sc) AAV as a vehicle for intra-articular gene delivery. Methods Conventional and scAAV vectors were used to infect lapine articular fibroblasts in culture to determine transduction efficiency, transgene expression levels, and nuclear trafficking. scAAV containing the cDNA for interleukin (IL)-1 receptor antagonist (Ra) was delivered to the joints of naive rabbits and those with IL-1 beta-induced arthritis. From lavage of the joint space, levels of transgenic expression and persistence were measured by enzyme-linked immunosorbent assay. infiltrating leukocytes were quantified using a hemocytometer. Results Transgene expression from scAAV had an earlier onset and was approximately 25-fold greater than conventional AAV despite the presence of similar numbers of viral genomes in the nuclei of infected cells. Fibroblasts transduced with scAAV produced amounts of IL1-Ra comparable to those transduced with adenoviral and lentiviral vectors. IL1-Ra was present in lavage fluid of most animals for 2 weeks in sufficient quantities to inhibit inflammation of the IL-1 beta-driven model. Once lost, neither Subsequent inflammatory events, nor re-ad ministration of the virus Could re-establish transgene expression. Conclusions scAAV-mediated intra-articular gene transfer is robust and similarly efficient in both normal and inflamed joints; the resulting transgenic expression is sufficient to achieve biological relevance in joints of human proportion. Copyright (C) 2009 John Wiley & Sons, Ltd.
Journal of Gene Medicine
"Intra-articular gene delivery and expression of interleukin-1Ra mediated by self-complementary adeno-associated virus" (2009). Faculty Bibliography 2000s. 1709.