title: Genome-wide microarray analyses identify the protein C receptor as a novel calcineurin/nuclear factor of activated T cells-dependent gene in vascular smooth muscle cell phenotypic modulation.
abstract: OBJECTIVE
  Calcineurin (Cn) and the nuclear factor of activated T cells (NFAT) family of transcription factors are critical in vascular smooth muscle cell (SMC) development and pathology. Here, we used a genomics approach to identify and validate NFAT gene targets activated during platelet-derived growth factor-BB (PDGF-BB)-induced SMC phenotypic modulation.
  
  
  RESULTS
  Genome-wide expression arrays were used to identify genes both (1) differentially activated in response to PDGF-BB and (2) whose differential expression was reduced by both the Cn inhibitor cyclosporin A and the NFAT inhibitor A-285222. The 20 most pharmacologically sensitive genes were validated by quantitative reverse transcription-polymerase chain reaction analysis of PDGF-BB-stimulated SMCs in the presence of Cn/NFAT inhibitors, including the VIVIT peptide. In all experiments, protein C receptor (PROCR) gene activation was reduced. We showed that PROCR expression was virtually absent in untreated, quiescent SMCs. PDGF-BB stimulation, however, induced significant PROCR promoter activation and downstream protein expression in a Cn/NFAT-dependent manner. Mutation of a species-conserved, NFAT binding motif significantly attenuated PDGF-BB-induced PROCR promoter activity, thereby distinguishing NFAT as the first PROCR transcriptional activator to date. Moreover, SMC PROCR expression was upregulated in the neointima as early as 7 days following acute vascular injury in rat carotid arteries.
  
  
  CONCLUSIONS
  We hereby report PROCR as a novel, NFAT-dependent gene that may be implicated in vascular restenosis and consequent inward remodeling.
authors: Department of Biomedical Engineering, University of Virginia, Charlottesville, VA 22908, USA.
authors: Monica Y
authors: MY
authors: Lee
authors: Sean M
authors: SM
authors: Garvey
authors: Marcia L
authors: ML
authors: Ripley
authors: Brian R
authors: BR
authors: Wamhoff
mesh_terms: D000818:Animals; D001483:Base Sequence; D000077214:Becaplermin; D001779:Blood Coagulation Factors / Q000235:genetics* / Q000378:metabolism; D019703:Calcineurin / Q000235:genetics* / Q000378:metabolism; D065095:Calcineurin Inhibitors; D020212:Carotid Artery Injuries / Q000378:metabolism; D002404:Catheterization / Q000009:adverse effects; D002478:Cells, Cultured; D016572:Cyclosporine / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D016678:Genome / Q000235:genetics*; D023421:Models, Animal; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D050778:NFATC Transcription Factors / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D020574:Proto-Oncogene Proteins c-sis; D011720:Pyrazoles / Q000494:pharmacology; D051381:Rats; D011956:Receptors, Cell Surface / Q000235:genetics* / Q000378:metabolism
chemical_list: C494244:A 285222; D001779:Blood Coagulation Factors; D065095:Calcineurin Inhibitors; D050778:NFATC Transcription Factors; D010982:Platelet-Derived Growth Factor; D020574:Proto-Oncogene Proteins c-sis; D011720:Pyrazoles; D011956:Receptors, Cell Surface; C091963:activated protein C receptor; D000077214:Becaplermin; D016572:Cyclosporine; D019703:Calcineurin