Journal of Molecular Medicine

, Volume 90, Issue 3, pp 331–342 | Cite as

Adenosine A2A receptor activation stimulates collagen production in sclerodermic dermal fibroblasts either directly and through a cross-talk with the cannabinoid system

  • Pietro Enea Lazzerini
  • Mariarita Natale
  • Elena Gianchecchi
  • Pier Leopoldo Capecchi
  • Cinzia Montilli
  • Stefania Zimbone
  • Monica Castrichini
  • Epifania Balistreri
  • Gianluca Ricci
  • Enrico Selvi
  • Estrella Garcia-Gonzalez
  • Mauro Galeazzi
  • Franco Laghi-Pasini
Original Article

Abstract

Systemic sclerosis (SSc) is a connective tissue disease characterised by exaggerated collagen deposition in the skin and visceral organs. Adenosine A2A receptor stimulation (A2Ar) promotes dermal fibrosis, while the cannabinoid system modulates fibrogenesis in vitro and in animal models of SSc. Moreover, evidence in central nervous system suggests that A2A and cannabinoid (CB1) receptors may physically and functionally interact. On this basis, we investigated A2Ar expression and function in modulating collagen biosynthesis from SSc dermal fibroblasts and analysed the cross-talk with cannabinoid receptors. In sclerodermic cells, A2Ar expression (RT-PCR, Western blotting) was evaluated together with the effects of A2A agonists and/or antagonists on collagen biosynthesis (EIA, Western blotting). Putative physical and functional interactions between the A2A and cannabinoid receptors were respectively assessed by co-immuno-precipitation and co-incubating the cells with the unselective cannabinoid agonist WIN55,212-2, and the selective A2A antagonist ZM-241385. In SSc fibroblasts, (1) the A2Ar is overexpressed and its occupancy with the selective agonist CGS-21680 increases collagen production, myofibroblast trans-differentiation, and ERK-1/2 phosphorylation; (2) the A2Ar forms an heteromer with the cannabinoid CB1 receptor; and (3) unselective cannabinoid receptor stimulation with a per se ineffective dose of WIN55,212-2, results in a marked anti-fibrotic effect after A2Ar blockage. In conclusion, A2Ar stimulation induces a pro-fibrotic phenotype in SSc dermal fibroblasts, either directly, and indirectly, by activating the CB1 cannabinoid receptor. These findings increase our knowledge of the pathophysiology of sclerodermic fibrosis also further suggesting a new therapeutic approach to the disease.

Keywords

Systemic sclerosis Dermal fibroblasts Fibrogenesis Adenosine A2A receptor Cannabinoid receptors 

References

  1. 1.
    Gabrielli A, Avvedimento EV, Krieg T (2009) Scleroderma. N Eng J Med 360:1989–2003CrossRefGoogle Scholar
  2. 2.
    Chan ES, Montesinos MC, Fernandez P, Desai A, Delano DL, Yee H, Reiss AB, Pillinger MH, Chen JF, Schwarzschild MA et al (2006) Adenosine A(2A) receptors play a role in the pathogenesis of hepatic cirrhosis. Br J Pharmacol 148:1144–1155PubMedCrossRefGoogle Scholar
  3. 3.
    Nakav S, Kachko L, Vorobiov M, Rogachev B, Chaimovitz C, Zlotnik M, Douvdevani A (2009) Blocking adenosine A2A receptor reduces peritoneal fibrosis in two independent experimental models. Nephrol Dial Transplant 24:2392–2399PubMedCrossRefGoogle Scholar
  4. 4.
    Chan ES, Fernandez P, Merchant AA, Montesinos MC, Trzaska S, Desai A, Tung CF, Khoa DN, Pillinger MH, Reiss AB et al (2006) Adenosine A2A receptors in diffuse dermal fibrosis: pathogenic role in human dermal fibroblasts and in a murine model of scleroderma. Arthritis Rheu 54:2632–2642CrossRefGoogle Scholar
  5. 5.
    Katebi M, Fernandez P, Chan ES, Cronstein BN (2008) Adenosine A2A receptor blockade or deletion diminishes fibrocyte accumulation in the skin in a murine model of scleroderma, bleomycin-induced fibrosis. Inflammation 3:299–303CrossRefGoogle Scholar
  6. 6.
    Chunn JL, Mohsenin A, Young HW, Lee CG, Elias JA, Kellems RE, Blackburn MR (2006) Partially adenosine deaminase-deficient mice develop pulmonary fibrosis in association with adenosine elevations. Am J Physiol Lung Cell Mol Physiol 290:L579–L587PubMedCrossRefGoogle Scholar
  7. 7.
    Fernández P, Trzaska S, Wilder T, Chiriboga L, Blackburn MR, Cronstein BN, Chan ES (2008) Pharmacological blockade of A2A receptors prevents dermal fibrosis in a model of elevated tissue adenosine. Am J Pathol 172:1675–1682PubMedCrossRefGoogle Scholar
  8. 8.
    Graham ES, Ashton JC, Glass M (2009) Cannabinoid receptors: a brief history and “what’s hot”. Front Biosci 14:944–957PubMedCrossRefGoogle Scholar
  9. 9.
    Caraceni P, Domenicali M, Giannone F, Bernardi M (2009) The role of the endocannabinoid system in liver diseases. Best Pract Res Clin Endocrinol Metab 23:65–77PubMedCrossRefGoogle Scholar
  10. 10.
    Michalski CW, Maier M, Erkan M, Sauliunaite D, Bergmann F, Pacher P, Batkai S, Giese NA, Giese T, Friess H et al (2008) Cannabinoids reduce markers of inflammation and fibrosis in pancreatic stellate cells. PLoS One 3:e1701PubMedCrossRefGoogle Scholar
  11. 11.
    Akhmetshina A, Dees C, Busch N, Beer J, Sarter K, Zwerina J, Zimmer A, Distler O, Schett G, Distler JH (2009) The cannabinoid receptor CB2 exerts antifibrotic effects in experimental dermal fibrosis. Arthritis Rheum 60:1129–1136PubMedCrossRefGoogle Scholar
  12. 12.
    Garcia-Gonzalez E, Selvi E, Balistreri E, Lorenzini S, Maggio R, Natale MR, Capecchi PL, Lazzerini PE, Bardelli M, Laghi-Pasini F, Galeazzi M (2009) Cannabinoids inhibit fibrogenesis in diffuse systemic sclerosis fibroblasts. Rheumatology 48:1050–1056PubMedCrossRefGoogle Scholar
  13. 13.
    Balistreri E, Garcia-Gonzalez E, Selvi E, Akhmetshina A, Palumbo K, Lorenzini S, Maggio R, Lucatelli M, Galeazzi M, Distler JW (2011) The cannabinoid WIN55, 212-2 abrogates dermal fibrosis in scleroderma bleomycin model. Ann Rheum Dis 70:695–699PubMedCrossRefGoogle Scholar
  14. 14.
    Servettaz A, Kavian N, Nicco C, Deveaux V, Chéreau C, Wang A, Zimmer A, Lotersztajn S, Weill B, Batteux F (2010) Targeting the cannabinoid pathway limits the development of fibrosis and autoimmunity in a mouse model of systemic sclerosis. Am J Pathol 177:187–196PubMedCrossRefGoogle Scholar
  15. 15.
    Ferré S, Goldberg SR, Lluis C, Franco R (2009) Looking for the role of cannabinoid receptor heteromers in striatal function. Neuropharmacology 56:226–234PubMedCrossRefGoogle Scholar
  16. 16.
    Carriba P, Ortiz O, Patkar K, Justinova Z, Stroik J, Themann A, Müller C, Woods AS, Hope BT, Ciruela F et al (2007) Striatal adenosine A2A and cannabinoid CB1 receptors form functional heteromeric complexes that mediate the motor effects of cannabinoids. Neuropsychopharmacology 32:2249–2259PubMedCrossRefGoogle Scholar
  17. 17.
    Subcommittee for Sclerodermia Criteria of the American Rheumatism Association Diagnostic and Therapeutic Criteria Committee (1980) Preliminary criteria for the classification of systemic sclerosis (scleroderma). Arthritis Rheum 23:581–590CrossRefGoogle Scholar
  18. 18.
    Abraham DJ, Eckes B, Rajkumar V, Krieg T (2007) New developments in fibroblast and myofibroblast biology: implications for fibrosis and scleroderma. Curr Rheumatol Rep 9:136–143PubMedCrossRefGoogle Scholar
  19. 19.
    Chen Y, Shi-Wen X, van Beek J, Kennedy L, McLeod M, Renzoni EA, Bou-Gharios G, Wilcox-Adelman S, Goetinck PF, Eastwood M et al (2005) Matrix contraction by dermal fibroblasts requires transforming growth factor-beta/activin-linked kinase 5, heparansulfate-containing proteoglycans, and MEK/ERK: insights into pathological scarring in chronic fibrotic disease. Am J Pathos 167:1699–1711CrossRefGoogle Scholar
  20. 20.
    Baroni SS, Santillo M, Bevilacqua F, Luchetti M, Spadoni T, Mancini M, Fraticelli P, Sambo P, Funaro A, Kazlauskas A et al (2006) Stimulatory autoantibodies to the PDGF receptor in systemic sclerosis. N Engl J Med 354:2667–2676PubMedCrossRefGoogle Scholar
  21. 21.
    Pertwee RG (1999) Pharmacology of cannabinoid receptor ligands. Curr Med Chem 6:635–664PubMedGoogle Scholar
  22. 22.
    Haskó G, Linden J, Cronstein B, Pacher P (2008) Adenosine receptors: therapeutic aspects for inflammatory and immune diseases. Nat Rev Drug Discov 7:759–770PubMedCrossRefGoogle Scholar
  23. 23.
    Nguyen DK, Montesinos MC, Williams AJ, Kelly M, Cronstein BN (2003) Th1 cytokines regulate adenosine receptors and their downstream signaling elements in human microvascular endothelial cells. J Immunol 171:3991–3998PubMedGoogle Scholar
  24. 24.
    Capecchi PL, Camurri A, Pompella G, Mazzola A, Maccherini M, Diciolla F, Lazzerini PE, Abbracchio MP, Laghi-Pasini F (2005) Upregulation of A2A adenosine receptor expression by TNF-alpha in PBMC of patients with CHF: a regulatory mechanism of inflammation. J Card Fail 11:67–73PubMedCrossRefGoogle Scholar
  25. 25.
    Fortin A, Harbour D, Fernandes M, Borgeat P, Bourgoin S (2006) Differential expression of adenosine receptors in human neutrophils: up-regulation by specific Th1 cytokines and lipopolysaccharide. J Leukoc Biol 79:574–585PubMedCrossRefGoogle Scholar
  26. 26.
    Morello S, Ito K, Yamamura S, Lee KY, Jazrawi E, Desouza P, Barnes P, Cicala C, Adcock IM (2006) IL-1 beta and TNF-alpha regulation of the adenosine receptor (A2A) expression: differential requirement for NF-kappa B binding to the proximal promoter. J Immunol 177:7173–7183PubMedGoogle Scholar
  27. 27.
    Varani K, Massara A, Vincenzi F, Tosi A, Padovan M, Trotta F, Borea PA (2009) Normalization of A2A and A3 adenosine receptor up-regulation in rheumatoid arthritis patients by treatment with anti-tumor necrosis factor alpha but not methotrexate. Arthritis Rheum 60:2880–2891PubMedCrossRefGoogle Scholar
  28. 28.
    Howlett AC, Barth F, Bonner TI, Cabral G, Casellas P, Devane WA, Felder CC, Herkenham M, Mackie K, Martin BR et al (2002) International Union of Pharmacology. XXVII. Classification of cannabinoid receptors. Pharmacol Rev 54:161–202PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Pietro Enea Lazzerini
    • 1
  • Mariarita Natale
    • 1
  • Elena Gianchecchi
    • 1
  • Pier Leopoldo Capecchi
    • 1
  • Cinzia Montilli
    • 1
  • Stefania Zimbone
    • 1
  • Monica Castrichini
    • 1
  • Epifania Balistreri
    • 2
  • Gianluca Ricci
    • 1
  • Enrico Selvi
    • 2
  • Estrella Garcia-Gonzalez
    • 2
  • Mauro Galeazzi
    • 2
  • Franco Laghi-Pasini
    • 1
  1. 1.Department of Clinical Medicine and Immunological Sciences, Division of Clinical ImmunologyUniversity of SienaSienaItaly
  2. 2.Department of Clinical Medicine and Immunological Sciences, Division of RheumatologyUniversity of SienaSienaItaly

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