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Genomics approach using calcineurin/NFAT inhibitors to identify novel effectors of vascular smooth muscle phenotypic modulation
Author(s) -
Lee Monica Y,
Garvey Sean M,
Wamhoff Brian R
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.980.2
Subject(s) - nfat , calcineurin , phenotype , downregulation and upregulation , vascular smooth muscle , biology , platelet derived growth factor receptor , gene , microbiology and biotechnology , transcription factor , cancer research , growth factor , genetics , receptor , endocrinology , medicine , smooth muscle , transplantation
Calcineurin (Cn) and the NFAT family of transcription factors are critical in vascular smooth muscle cell (SMC) development and pathology. We used a genomics approach to identify novel transcriptional targets of NFAT in SMC phenotypic modulation. SMC phenotypic modulation was induced by treatment with platelet‐derived growth factor‐BB (PDGF). PDGF‐induced proliferation was blocked by either the Cn inhibitor, cyclosporin A (CsA), or the NFAT inhibitor, A285222. Genome‐wide expression arrays were used to identify genes in rat aortic SMCs both 1) differentially expressed in response to PDGF at 3 hours and 2) whose differential expression was reduced by both CsA and A285222; genes theoretically positively regulated by Cn/NFAT. The top 20 PDGF‐upregulated probe sets sensitive to both drugs corresponded to 17 genes. 15 of these genes were expressed at higher levels in intact rat arteries compared to visceral smooth muscle tissues. For validation, SMCs were treated at 10 time‐points across 24 hours. Real‐time RTPCR showed that CsA and A285222 reduced the early upregulation of DSCR1 (Cn regulator), DUSP1 (MAPK inactivator), and PROCR (protein C receptor) more than 50% compared to vehicle controls. Intriguingly, the PROCR promoter contains a highly conserved NFAT site at the ‐8 position. We report PROCR and DUSP1 as novel NFAT‐dependent early genes in VSMC phenotypic modulation. AHA predoc APS postdoc NIH HL81682 AHA SDG