Adenosine A2A receptor activation stimulates collagen production in sclerodermic dermal fibroblasts either directly and through a cross-talk with the cannabinoid system.
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.
Department of Clinical Medicine and Immunological Sciences, Division of Clinical Immunology, University of Siena, Siena, Italy.
Pietro Enea
PE
Lazzerini
Mariarita
M
Natale
Elena
E
Gianchecchi
Pier Leopoldo
PL
Capecchi
Cinzia
C
Montilli
Stefania
S
Zimbone
Monica
M
Castrichini
Epifania
E
Balistreri
Gianluca
G
Ricci
Enrico
E
Selvi
Estrella
E
Garcia-Gonzalez
Mauro
M
Galeazzi
Franco
F
Laghi-Pasini
D000328:Adult; D000368:Aged; D002186:Cannabinoids / Q000378:metabolism / Q000494:pharmacology; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis*; D020405:Dermis / Q000166:cytology / Q000378:metabolism / Q000503:physiopathology; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism* / Q000473:pathology; D005355:Fibrosis / Q000378:metabolism / Q000503:physiopathology; D006801:Humans; D008297:Male; D008875:Middle Aged; D010766:Phosphorylation; D043705:Receptor, Adenosine A2A / Q000235:genetics / Q000378:metabolism*; D043884:Receptor, Cannabinoid, CB1 / Q000235:genetics / Q000378:metabolism*; D012595:Scleroderma, Systemic / Q000378:metabolism / Q000503:physiopathology*; D015398:Signal Transduction; D015854:Up-Regulation
D002186:Cannabinoids; D043705:Receptor, Adenosine A2A; D043884:Receptor, Cannabinoid, CB1; D003094:Collagen