POS0373 LIRAGLUTIDE HAS POTENT ANTI-INFLAMMATORY AND ANTI-CATABOLIC IN VITRO ACTIVITIES IN OSTEOARTHRITIS

Background: Osteoarthritis (OA) is an age-related joint disease which provokes chronic pain and limits mobility. The disease progression is associated with inflammatory responses and cartilage degradation. Both chondrocytes, the only cell type present in cartilage, and macrophages from the synovium, play a major role in OA pathophysiology. Liraglutide is a Glucagon-Like Peptide-1 Receptor (GLP-1R) agonist widely prescribed for the treatment of type 2 diabetes. Interestingly, anti-inflammatory properties of the GLP-1 pathway have been reported in various diseases outside diabetes. Objectives: We evaluated the anti-inflammatory and anti-catabolic effects of Liraglutide in two in vitro models relevant to OA by evaluating surrogate markers of inflammation, cartilage matrix proteolysis and differentiation. Methods: Lipopolysaccharide (LPS)-stimulated murine Raw 264.7 macrophages were treated with 10 concentrations (6.6nM-3.4µM) of Liraglutide for 24h. Anti-inflammatory activity was evaluated by the production of nitric oxide (NO) and prostaglandin E2 (PGE2) using Griess reaction and ELISA, respectively. Interleukin 1β (IL-1β)-stimulated mouse articular chondrocytes were treated with Liraglutide (6.6nM-3.4µM) for 24h. Production of IL-6, matrix metalloproteinase-3 (MMP-3) and glycosaminoglycans (GAG) was measured by ELISA and GAG assay, respectively. RTqPCR analyses were performed with three selected concentrations of Liraglutide (13.3nM, 53.1nM and 1.7µM) on both cell types to assess the expression of a panel of genes related to inflammation (IL-6, TNF, iNOS), M1/M2 macrophage phenotype (MCP-1, CD38, ERG-2), catabolism (MMP-13, ADAMTS-5) and differentiation (Sox9, Col2a1, Acan). Results: Liraglutide induced a dose-dependent inhibition of the LPS-induced production of NO (IC50=45nM) and PGE2 (IC50=54nM) in macrophages. Moreover, IL-6 and TNF gene expressions were significantly and dose-dependently decreased in Raw 264.7 cells treated with Liraglutide compared to LPS alone. Interestingly, there was a significant dose-dependent reduction of MCP-1 and CD38 (M1 marker) gene expression in cells treated with the 3 doses of Liraglutide compared to LPS alone while we observed a dose-dependent increase of ERG-2 (M2 marker) gene expression induced by Liraglutide. Liraglutide significantly dose-dependently reduced the IL-1β-induced release of IL-6 (IC50=38nM), MMP-3 (IC50=56nM) and GAG (IC50=47nM) in chondrocytes. Additionally, Liraglutide treatment dose-dependently decreased the IL-1β-induced gene expression of iNOS, MMP-13 and ADAMTS-5. Finally, IL-1β decreased gene expression of Sox9, Col2a1 and Acan differentiation markers, which was rescued in a dose-dependent manner by Liraglutide (Table 1). Conclusion: A shift in M1/M2 macrophage phenotype and the inhibition of chondrocyte expression of several mediators involved in inflammation and cartilage degradation explain, at least in part, our previous results from rodent osteoarthritis models that showed an analgesic, anti-inflammatory and anti-degradative effect of Liraglutide. The fact that Liraglutide is already safely prescribed in another indication allows us to foresee a first trial in humans in the short term. Acknowledgements: All the people who contributed to the InOsteo project: the members of 4P-Pharma, INSERM UMRS_938 research team, SATT Lutech and Sorbonne University Disclosure of Interests: Francis Berenbaum Consultant of: Boehringer, Bone Therapeutics, CellProthera, Expanscience, Galapagos, Gilead, GSK, Merck Sereno, MSD, Nordic, Novartis, Pfizer, Regulaxis, Roche, Sandoz, Sanofi, Servier, UCB, Peptinov, 4P Pharma, 4Moving Biotech, Grant/research support from: TRB Chemedica, Coralie Meurot Employee of: 4P-Pharma, Laure Sudre Employee of: 4P-Pharma, Keren Bismuth Employee of: 4P-Pharma, Revital Rattenbach Shareholder of: 4P-Pharma, Employee of: 4P-Pharma, Patrice Denefle Speakers bureau: 4P-Pharma, Consultant of: 4P-Pharma, Pierre Fabre, Mimetas, Employee of: 4P-Pharma, Celine Martin Employee of: 4P-Pharma, Claire Jacques: None declared