Title

Intra-Articular Gene Delivery And Expression Of Interleukin-1Ra Mediated By Self-Complementary Adeno-Associated Virus

Keywords

Adeno-associated virus; Arthritis; Gene therapy; Interleukin-1; Interleukin-1 receptor antagonist

Abstract

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 naïve rabbits and those with IL-1β-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-1β-driven model. Once lost, neither subsequent inflammatory events, nor re-administration 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 © 2009 John Wiley & Sons, Ltd.

Publication Date

10-2-2009

Publication Title

Journal of Gene Medicine

Volume

11

Issue

7

Number of Pages

605-614

Document Type

Article

Personal Identifier

scopus

DOI Link

https://doi.org/10.1002/jgm.1334

Socpus ID

68949205604 (Scopus)

Source API URL

https://api.elsevier.com/content/abstract/scopus_id/68949205604

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