AAV-mediated Overexpression of IL-10 Mitigates the Inflammatory Cascade in Stimulated Equine Chondrocyte Pellets

Title:AAV-mediated Overexpression of IL-10 Mitigates the Inflammatory Cascade in Stimulated Equine Chondrocyte Pellets

Volume: 18 Issue: 3

Author(s): Kyla F. Ortved*, Laila Begum, Darko Stefanovski and Alan J. Nixon

Affiliation:

• Department of Clinical Sciences, Cornell University, Ithaca, NY 14853,United States

Keywords: Osteoarthritis, Gene therapy, AAV, IL-10, Immunomodulation, Chondrocyte pellets, Cytokine.

Abstract: Background: Following joint trauma, a posttraumatic inflammatory cascade drives degeneration of the joint. We aimed to assess whether transduction of chondrocytes with AAV5 overexpressing the immunomodulatory cytokine IL-10 would have protective effects in pellet cultures stimulated with IL-1β.

Methods: Chondrocytes were isolated from 3 healthy horses and were transduced with AAV5-IL-10 at a dose of 1 x 105vg/cell. Chondrocyte pellets were formed by centrifugation and were stimulated with IL-1β starting 48 hours following transduction. After 2, 6 and 14 days in culture, supernatants were collected for cytokine analysis and RNA was isolated from cells for gene expression analysis. Pellets were also collected for biochemical analysis.

Results: Transduction of chondrocytes led to significant increases in IL-10 expression. IL-10 expression was further enhanced by IL-1β stimulation. IL-10 overexpression led to significantly decreased expression of IL-1β and ADAMTS4. PGE2 synthesis was also significantly decreased. IL-1β mediated suppression of GAG synthesis was not rescued by IL-10.

Conclusions: Overexpression of IL-10 modulates the inflammatory response in chondrocytes, which may mitigate some of the deleterious effects of pro-inflammatory cascades in the posttraumatic joint. AAV5-IL-10 led to efficient and sustained overexpression of IL-10 in chondrocytes and could represent a viable treatment option for preventing osteoarthritis.

Cite this article as:

Ortved F. Kyla *, Begum Laila, Stefanovski Darko and Nixon J. Alan , AAV-mediated Overexpression of IL-10 Mitigates the Inflammatory Cascade in Stimulated Equine Chondrocyte Pellets, Current Gene Therapy 2018; 18 (3) . https://dx.doi.org/10.2174/1566523218666180510165123