The Immunosuppressant FTY720 (Fingolimod) enhances Glycosaminoglycan depletion in articular cartilage

BMC Musculoskeletal Disorders

Winfried B Graninger¹  Josef Hermann¹  Marie-Luise Kremser¹  Daniela Setznagl¹  Florentine C Fuerst¹  Thomas Ortner¹  Hannes Angerer¹  Martin H Stradner¹ 

[1] Division of Rheumatology and Immunology, Medical University of Graz, Austria

关键词: glycosaminoglycan;    inducible nitric oxide synthase;    tumor necrosis factor-α;    interleukin-1β;    FTY720;    fingolimod;    chondrocyte;   

Others  :  1150952
DOI  :  10.1186/1471-2474-12-279

 received in 2011-05-30, accepted in 2011-12-12,  发布年份 2011

【 摘 要 】

Background

FTY720 (Fingolimod) is a novel immunosuppressive drug investigated in clinical trials for organ transplantation and multiple sclerosis. It acts as a functional sphingosine-1-phosphate (S1P) receptor antagonist, thereby inhibiting the egress of lymphocytes from secondary lymphoid organs. As S1P is able to prevent IL-1beta induced cartilage degradation, we examined the direct impact of FTY720 on cytokine induced cartilage destruction.

Methods

Bovine chondrocytes were treated with the bioactive phosphorylated form of FTY720 (FTY720-P) in combination with IL-1beta or TNF-alpha. Expression of MMP-1,-3.-13, iNOS and ADAMTS-4,-5 and COX-2 was evaluated using quantitative real-time PCR and western blot. Glycosaminoglycan depletion from cartilage explants was determined using a 1,9-dimethylene blue assay and safranin O staining.

Results

FTY720-P significantly reduced IL-1beta and TNF-alpha induced expression of iNOS. In contrast FTY720-P increased MMP-3 and ADAMTS-5 mRNA expression. Furthermore depletion of glycosaminoglycan from cartilage explants by IL-1beta and TNF-alpha was significantly enhanced by FTY720-P in an MMP-3 dependent manner.

Conclusions

Our results suggest that FTY720 may enhance cartilage degradation in pro-inflammatory environment.

【 授权许可】

   
2011 Stradner et al; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

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【 参考文献 】

• [1]Tedesco-Silva H, Szakaly P, Shoker A, Sommerer C, Yoshimura N, Schena FP, Cremer M, Hmissi A, Mayer H, Lang P: FTY720 versus mycophenolate mofetil in de novo renal transplantation: six-month results of a double-blind study. Transplantation 2007, 84:885-892.
• [2]Kappos L, Radue EW, O'Connor P, Polman C, Hohlfeld R, Calabresi P, Selmaj K, Agoropoulou C, Leyk M, Zhang-Auberson L, et al.: A placebo-controlled trial of oral fingolimod in relapsing multiple sclerosis. N Engl J Med 362:387-401.
• [3]Cohen JA, Barkhof F, Comi G, Hartung HP, Khatri BO, Montalban X, Pelletier J, Capra R, Gallo P, Izquierdo G, et al.: Oral fingolimod or intramuscular interferon for relapsing multiple sclerosis. N Engl J Med 362:402-415.
• [4]Brinkmann V, Wilt C, Kristofic C, Nikolova Z, Hof RP, Chen S, Albert R, Cottens S: FTY720: dissection of membrane receptor-operated, stereospecific effects on cell migration from receptor-independent antiproliferative and apoptotic effects. Transplant Proc 2001, 33:3078-3080.
• [5]Takabe K, Paugh S, Milstien S, Spiegel S: "Inside-out" signaling of sphingosine-1-phosphate: therapeutic targets. Pharmacol Rev 2008, 60:181-195.
• [6]Graler MH, Goetzl EJ: The immunosuppressant FTY720 down-regulates sphingosine 1-phosphate G protein-coupled receptors. FASEB J 2004, 18:551-553.
• [7]Matloubian M, Lo CG, Cinamon G, Lesneski MJ, Xu Y, Brinkmann V, Allende ML, Proia RL, Cyster JG: Lymphocyte egress from thymus and peripheral lymphoid organs is dependent on S1P receptor 1. Nature 2004, 427:355-360.
• [8]Kikuta J, Iwai K, Saeki Y, Ishii M: S1P-targeted therapy for elderly rheumatoid arthritis patients with osteoporosis. Rheumatol Int 2010, 31:967-9.
• [9]Wang F, Tan W, Guo D, He S: Reduction of CD4 positive T cells and improvement of pathological changes of collagen-induced arthritis by FTY720. Eur J Pharmacol 2007, 573:230-240.
• [10]Matsuura M, Imayoshi T, Okumoto T: Effect of FTY720, a novel immunosuppressant, on adjuvant- and collagen-induced arthritis in rats. Int J Immunopharmacol 2000, 22:323-331.
• [11]Stradner MH, Hermann J, Angerer H, Setznagl D, Sunk IG, Windhager R, Graninger WB: Spingosine-1-phosphate stimulates proliferation and counteracts interleukin-1 induced nitric oxide formation in articular chondrocytes. Osteoarthritis Cartilage 2008, 16:305-311.
• [12]Kim MK, Lee HY, Kwak JY, Park JI, Yun J, Bae YS: Sphingosine-1-phosphate stimulates rat primary chondrocyte proliferation. Biochem Biophys Res Commun 2006, 345:67-73.
• [13]Masuko K, Murata M, Nakamura H, Yudoh K, Nishioka K, Kato T: Sphingosine-1-phosphate attenuates proteoglycan aggrecan expression via production of prostaglandin E2 from human articular chondrocytes. BMC Musculoskeletal Disorders 2007, 8:29. BioMed Central Full Text
• [14]Primer3 on the WWW for general users and for biologist programmers [http://frodo.wi.mit.edu/cgi-bin/primer3/primer3_www.cgi] webcite
• [15]Bustin SA, Beaulieu JF, Huggett J, Jaggi R, Kibenge FS, Olsvik PA, Penning LC, Toegel S: MIQE précis: Practical implementation of minimum standard guidelines for fluorescence-based quantitative real-time PCR experiments. BMC Mol Biol 2010, 11:74. BioMed Central Full Text
• [16]Richard W, Farndale CAS, Barrett AlanJ: A Direct Spectrophotometric Microassay for Sulfated Glucosaminoglycans in Cartilage Cultures. Connective Tissue Research 1982, 9:247-248.
• [17]Camplejohn KL, Allard SA: Limitations of safranin 'O' staining in proteoglycan-depleted cartilage demonstrated with monoclonal antibodies. Histochemistry 1988, 89:185-188.
• [18]Tsunemi S, Iwasaki T, Kitano S, Imado T, Miyazawa K, Sano H: Effects of the novel immunosuppressant FTY720 in a murine rheumatoid arthritis model. Clin Immunol 2010, 136:197-204.
• [19]Angyal A, Egelston C, Kobezda T, Olasz K, László A, Glant T, Mikecz K: Development of proteoglycan-induced arthritis depends on T cell-supported autoantibody production, but does not involve significant influx of T cells into the joints. Arthritis Res Ther 2010, 12:R44. BioMed Central Full Text
• [20]Brinkmann V, Davis MD, Heise CE, Albert R, Cottens S, Hof R, Bruns C, Prieschl E, Baumruker T, Hiestand P, et al.: The immune modulator FTY720 targets sphingosine 1-phosphate receptors. J Biol Chem 2002, 277:21453-21457.
• [21]Xin C, Ren S, Kleuser B, Shabahang S, Eberhardt W, Radeke H, Schafer-Korting M, Pfeilschifter J, Huwiler A: Sphingosine 1-Phosphate Cross-activates the Smad Signaling Cascade and Mimics Transforming Growth Factor-{beta}-induced Cell Responses. J Biol Chem 2004, 279:35255-35262.
• [22]Clancy R, Rediske J, Koehne C, Stoyanovsky D, Amin A, Attur M, Iyama K, Abramson SB: Activation of stress-activated protein kinase in osteoarthritic cartilage: evidence for nitric oxide dependence. Osteoarthritis and Cartilage 2001, 9:294.
• [23]Blanco FJ, Ochs RL, Schwarz H, Lotz M: Chondrocyte apoptosis induced by nitric oxide. Am J Pathol 1995, 146:75-85.
• [24]Van de Loo FAJ, Arntz OJ, Van den Berg WB: Effect of Interleukin-1 and Leukemia Inhibitory Factor on Chondrocyte Metabolism in Articular Cartilage from Normal and Interleukin-6-deficient Mice: Role of Nitric Oxide and IL-6 in the Suppression of Proteoglycan Synthesis. Cytokine 1997, 9:453.
• [25]Cao M, Westerhausen-Larson A, Niyibizi CKK, Georgescu HI, Rizzo CF, Hebda PA, Stefanovic-Racic M, CH E: Nitric oxide inhibits the synthesis of type-II collagen without altering Col2A1 mRNA abundance: prolyl hydroxylase as a possible target. Biochem J 1997, (Pt 1):305-310.
• [26]van den Berg WB, van de Loo F, Joosten LAB, Arntz OJ: Animal models of arthritis in NOS2-deficient mice. Osteoarthritis and Cartilage 1999, 7:413.
• [27]Matsumoto N, Kohno T, Uchida S, Shimizu T, Kusumoto H, Hirose R, Yanada K, Kurio W, Yamaguchi S, Watabe K, et al.: Preparation of antibodies against a novel immunosuppressant, FTY720, and development of an enzyme immunoassay for FTY720. Bioorg Med Chem 2006, 14:4182-4192.