Arthritis & Rheumatism, Volume 62,
November 2010 Abstract Supplement
Abstracts of the American College of
Rheumatology/Association of Rheumatology Health Professionals
Annual Scientific Meeting
Atlanta, Georgia November 6-11, 2010.
The Effects of Manzamine A on Rheumatoid Arthritis Synoviocyte Cytokine Secretion.
Chong1, Kyle, Klosowska1, Karolina, Woods2, James M.
Manzamine A (MZA) is a marine alkaloid isolated from sponges on the ocean floor known to have anti-inflammatory properties on activated macrophages. The complete synthesis of this complex compound has recently been reported. Rheumatoid Arthritis (RA) is characterized by increased production of pro-inflammatory cytokines by a variety of synovial cells found in the synovial tissue (ST) including fibroblast-like synoviocytes (FLS). We hypothesized that MZA would decrease the levels of pro-inflammatory cytokines secreted by RA cell types.
RA FLS and RA synovial tissue (ST) explants collected from patients with IRB approval were treated with MZA (10 mM, 1 mM, 0.1 mM, 0.05 mM, and 0.01 mM MZA) or vehicle control, to observe for alterations in pro-inflammatory cytokine production and to observe any cytotoxic effects. Condition media (CM), with or without IL-1b stimulation was collected at various time points. Toxicity was determined using trypan blue exclusion while cytokine levels were analyzed using antibody microarrays and ELISAs.
Toxicity studies on RA FLS suggest that concentrations of 10 mM, 1.0 mM and 0.1 mM MZA had no toxic effects after 24 hours of incubation. However, after 48 and 72 hours, the 10 mM and 1.0 mM MZA concentrations appear to significantly decrease FLS viability. Using an antibody microarray, 24 hour CM with 10 mM MZA appeared to decrease the production of several pro-inflammatory mediators in both RA FLS and RA ST. Proteins decreased include monocyte chemoattractant protein (MCP)-1 (CCL2), interleukin (IL)-8 (CXCL8), growth related oncogene (Gro)-a (CXCL1), regulated on activation of normal T cell expressed and secreted (RANTES/CCL5), IL-6, and epithelial neutrophil activating peptide (ENA)-78 (CXCL5). To confirm data obtained from the cytokine microarray analysis, the level of protein expression was further examined by ELISA. At 24 hours, 10 mM, 1.0 mM and 0.1 mM MZA significantly decreased the production of MCP-1 by 57%, 37 % and 44% respectively (n=5; p<0.05). Additionally 0.1 mM MZA decreased MCP-1 by 43% at 48 hours (n=5; p<0.05). MCP-1 production, when stimulated with IL-1b, was significantly decrease with 10 mM, 1.0 mM, 0.1 mM, 0.05 mM and 0.01 mM MZA by 87%, 84%, 57%, 62%, and 67%, respectively, at 24 hours (n=5; p<0.05). RANTES production was significantly decreased with 10 mM and 1.0 mM MZA by 79% and 69%, respectively, at 24 hours, and also by 54% with 0.1 mM MZA at 72 hours (n=5; p<0.05). IL-1b stimulated RANTES production was significantly decreased with 10 mM and 1.0 mM MZA by 86% and 75%, respectively, at 24 hours, and also by 59% with 0.1 mM MZA after 72 hours (n=5; p<0.05). Also, IL-1b stimulated Gro-a production was significantly decreased by 0.01 mM MZA by 26% after 24 hours (n=5; p<0.05).
While some concentrations of MZA applied appeared toxic to RA FLS, we identified a number of significant effects of MZA at non-toxic concentrations and time points. Our data suggest that MZA decreases RA-associated inflammation by reducing pro-inflammatory cytokines and appears to have solid pharmacological potential to serve as a novel therapeutic in RA therapy.
To cite this abstract, please use the following information:
Chong, Kyle, Klosowska, Karolina, Woods, James M.; The Effects of Manzamine A on Rheumatoid Arthritis Synoviocyte Cytokine Secretion. [abstract]. Arthritis Rheum 2010;62 Suppl 10 :1131