Dartmouth College Dartmouth Digital Commons Dartmouth Scholarship Faculty Work 12-1-2008 Retinoid X Receptor and Peroxisome Proliferator-Activated Receptor-Gamma Agonists Cooperate to Inhibit Matrix Metalloproteinase Gene Expression Peter S. Burrage Dartmouth College Adam C. Schmucker Dartmouth College Yanqing Ren Dartmouth College Michael B. Sporn Dartmouth College Constance E. Brinckerhoff Dartmouth College Follow this and additional works at: https://digitalcommons.dartmouth.edu/facoa Part of the Genetic Processes Commons, Medical Biochemistry Commons, and the Rheumatology Commons Dartmouth Digital Commons Citation Burrage, Peter S.; Schmucker, Adam C.; Ren, Yanqing; Sporn, Michael B.; and Brinckerhoff, Constance E., "Retinoid X Receptor and Peroxisome Proliferator-Activated Receptor-Gamma Agonists Cooperate to Inhibit Matrix Metalloproteinase Gene Expression" (2008). 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Available online http://arthritis-research.com/content/10/6/R139 ResearchVol 10 No 6 article Open Access Retinoid X receptor and peroxisome proliferator-activated receptor-gamma agonists cooperate to inhibit matrix metalloproteinase gene expression Peter S Burrage1*, Adam C Schmucker1*, Yanqing Ren1, Michael B Sporn2 and Constance E Brinckerhoff1,3 1Department of Biochemistry, Dartmouth Medical School, North College Street, 7200 Vail Building, Hanover, NH 03755, USA 2Department of Pharmacology, Dartmouth Medical School, North College Street, 7650 Remsen Hall, Hanover, NH 03755, USA 3Department of Medicine, Dartmouth Medical School, 1 Medical Center Drive, Lebanon NH 03756, USA * Contributed equally Corresponding author: Constance E Brinckerhoff, [email protected] Received: 8 Sep 2008 Revisions requested: 9 Oct 2008 Revisions received: 6 Nov 2008 Accepted: 1 Dec 2008 Published: 1 Dec 2008 Arthritis Research & Therapy 2008, 10:R139 (doi:10.1186/ar2564) This article is online at: http://arthritis-research.com/content/10/6/R139 © 2008 Burrage et al.; licensee BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Introduction We recently described the ability of retinoid X Results Rosiglitazone inhibited MMP-1 and MMP-13 receptor (RXR) ligand LG100268 (LG268) to inhibit interleukin- expression in IL-1--treated SW-1353 cells at the mRNA and 1-beta (IL-1-)-driven matrix metalloproteinase-1 (MMP-1) and heterogeneous nuclear RNA levels and blunted IL-1--induced MMP-13 gene expression in SW-1353 chondrosarcoma cells. collagen destruction in vitro. Combining LG268 and Other investigators have demonstrated similar effects in rosiglitazone had an additive inhibitory effect on MMP-1 and chondrocytes treated with rosiglitazone, a ligand for peroxisome MMP-13 transcription and collagenolysis. IL-1- inhibited proliferator-activated receptor-gamma (PPAR), for which RXR luciferase expression in the MMP reporter assay, but is an obligate dimerization partner. The goals of this study were rosiglitazone and LG268 had no effect. ChIP indicated that to evaluate the inhibition of IL-1--induced expression of MMP- treatment with IL-1-, but not LG268 and rosiglitazone, 1 and MMP-13 by combinatorial treatment with RXR and increased PPAR at the proximal promoters of both MMPs. PPAR ligands and to investigate the molecular mechanisms of Finally, rosiglitazone or LG268 induced 'cross-SUMOylation' of this inhibition. both the target receptor and its binding partner, and IL-1--alone had no effect on SUMOylation of RXR and PPAR but Methods We used real-time reverse transcription-polymerase antagonized the ligand-induced SUMOylation of both receptors. chain reaction to measure LG268- and rosiglitazone-mediated inhibition of MMP gene transcription in IL-1--treated SW-1353 chondrosarcoma cells. An in vitro collagen destruction assay Conclusions The PPAR and RXR ligands rosiglitazone and was a functional readout of MMP collagenolytic activity. LG268 may act through similar mechanisms, inhibiting MMP-1 Luciferase reporter assays tested the function of a putative and MMP-13 transcription. Combinatorial treatment activates regulatory element in the promoters of MMP-1 and MMP-13, each partner of the RXR:PPAR heterodimer and inhibits IL-1-- and chromatin immunoprecipitation (ChIP) assays detected induced expression of MMP-1 and MMP-13 more effectively PPAR and changes in histone acetylation at this site. Post- than either compound alone. We conclude that the efficacy of translational modification of RXR and PPAR by small ubiquitin- combined treatment with lower doses of each drug may like modifier (SUMO) was assayed with immunoprecipitation minimize potential side effects of treatment with these and Western blot. compounds. AP-1: activator protein-1; ChIP: chromatin immunoprecipitation; DMEM: Dulbecco's modified Eagle's medium; DR-1: direct repeat-1; ECM: extracel- lular matrix; FBS: fetal bovine serum; FXR: farnesoid X receptor; GAPDH: glyceraldehyde 3-phosphate dehydrogenase; HA: hemagglutinin; HA- PPAR: hemagglutinin-tagged peroxisome proliferator-activated receptor-gamma; HAT: histone acetyltransferase; HBSS: Hanks' balanced salt solu- tion; HDAC: histone deacetylase; hnRNA: heterogeneous nuclear RNA; IL-1: interleukin-1-beta; IP: immunoprecipitation; LG268: LG100268; LH: lactalbumin hydrosylate; LXR: liver X receptor; MMP: matrix metalloproteinase; MMPI: matrix metalloproteinase inhibitor; MSS: musculoskeletal syn- drome; NHR: nuclear hormone receptor; OA: osteoarthritis; PBS: phosphate-buffered saline; PCR: polymerase chain reaction; PPAR: peroxisome proliferator-activated receptor-gamma; PPRE: peroxisome proliferator-activated receptor-gamma response element; RA: rheumatoid arthritis; RT: reverse transcription; RXR: retinoid X receptor; SUMO: small ubiquitin-like modifier; TCA: trichloracetic acid. Page 1 of 16 (page number not for citation purposes) Arthritis Research & Therapy Vol 10 No 6 Burrage et al. Introduction ing the enzymatic activity of histone deacetylases (HDACs) The matrix metalloproteinases (MMPs) are a family of zinc- and histone acetyltransferases (HATs) [12]. HDAC activity dependent endopeptidases responsible for the degradation of results in a decrease in histone acetylation and a subsequent extracellular matrix (ECM) components. While low levels of decrease in transcriptional activity, whereas HAT activity leads these enzymes are required for the homeostatic ECM turnover to an increase in histone acetylation and a subsequent seen in wound healing, angiogenesis, and development, high increase in transcriptional activity [13]. levels have been implicated in the pathology of atherosclero- sis, tumor metastasis, and the arthritides. In the case of oste- Recent work has identified an anti-inflammatory role for PPAR oarthritis (OA) and rheumatoid arthritis (RA), members of the in chondrocytes when the receptor is activated by ligands collagenase subgroup of the MMPs, specifically MMP-1 and such as the thiazolidinedione compound rosiglitazone and the MMP-13, are particularly important in the progression of joint prostaglandin 15-Deoxy-12,14-prostaglandin J2 [2,14-16]. disease [1,2]. The ability to cleave the collagen triple helix is Notably, this anti-inflammatory effect of PPAR ligands unique to the collagenases, and the overexpression of MMP-1 extends to the inhibition of IL-1-induced expression of MMP- and MMP-13 in chondrocytes in response to proinflammatory 1 and MMP-13 in rabbit chondrocytes [2,17,18], and admin- cytokines such as interleukin-1-beta (IL-1) and tumor necro- istration of these compounds blunts the development of joint sis factor-alpha is critical in the pathogenesis of OA and RA disease in animal models of arthritis [18,19]. François and col- [1]. leagues [17] have proposed a mechanism to explain rosiglita- zone-mediated inhibition of IL-1-induced expression of rabbit Many efforts to design small-molecule inhibitors of MMP activ- MMP-1 that involves binding of the RXR:PPAR heterodimer ity (MMPIs) have succeeded in creating potent compounds; to a degenerate DR-1 site in the proximal (approximately -72 however, due to the highly conserved nature of the catalytic base pairs) region of the rabbit MMP-1 promoter. This DR-1 domain among family members, these compounds demon- site overlaps a binding site for the transcription factor activator strate significant inhibitory efficacy against multiple MMPs [3]. protein-1 (AP-1), which is largely responsible for the proinflam- This lack of specificity has been identified as the likely cause matory cytokine-induced upregulation of MMP-1 [20]. In this of the debilitating side effects observed in clinical trials with competitive binding model, binding of the RXR:PPAR het- these compounds which presented as a chronic musculoskel- erodimer to the DR-1 element precludes binding of AP-1 pro- etal syndrome (MSS) that was characterized by reduced teins to its site and thereby antagonizes the expression of mobility with joint pain and edema due to tendonitis and inflam- MMP-1. François and colleagues