title: Selective modulation of nuclear factor of activated T-cell function in restenosis by a potent bipartite peptide inhibitor.
abstract: BACKGROUND
  Nuclear factor of activated T-cells (NFAT) is importantly implicated in pathological cardiac remodeling and vascular lesion formation. NFAT functionality is mainly regulated by calcineurin, a Ca(2+)-dependent multi-effector phosphatase. Calcineurin inhibitors such as cyclosporine A (CsA) were shown to be effective in the treatment of restenosis and vascular inflammation but with adverse side effects.
  
  
  OBJECTIVE
  This prompted the design of more selective inhibitors such as VIVIT and inhibitors of NFAT-calcineurin association, which unfortunately have a poor potency precluding clinical use.
  
  
  RESULTS
  Here, we describe the rational design of a potent bipartite inhibitor of NFAT-calcineurin interaction, MCV1, which targets two separate calcineurin docking motifs. Modeling, site-directed mutagenesis, and functional studies demonstrated that MCV1 acts by allosteric modulation of calcineurin. Comparable to CsA, MCV1 prevents NFAT activation at nanomolar potency without impairing calcineurin phosphatase activity, nuclear factor-κB nuclear import, and general cell signaling. In contrast, CsA but not MCV1-activated basal level extracellular signal-regulated kinases activity and prevented nuclear import of calcineurin, independent of NFAT activation. In vivo MCV1 abrogated NFAT-mediated T-cell activation in a model of PMA-elicited peritonitis, whereas topical application of MCV1 markedly reduced neointima formation in a mouse model of restenosis.
  
  
  CONCLUSIONS
  We designed a bipartite NFAT inhibitor that is more potent than VIVIT and more selective than CsA. MCV1 constitutes not only a powerful tool to unravel NFAT function but also a potential drug candidate for the treatment of diseases implicating NFAT activation.
authors: Department of Pathology, University Hospital Maastricht, Debeijelaan 25, 6229 HX Maastricht, the Netherlands. erik.biessen@path.unimaas.nl
authors: Haixiang
authors: H
authors: Yu
authors: Ilze
authors: I
authors: Bot
authors: Karen
authors: K
authors: Sliedregt
authors: Xingfu
authors: X
authors: Xu
authors: Martine
authors: M
authors: Bot
authors: Sandra H
authors: SH
authors: van Heiningen
authors: Gijs A
authors: GA
authors: van der Marel
authors: Martin R
authors: MR
authors: Bennett
authors: Herman
authors: H
authors: Overkleeft
authors: Theo J C
authors: TJ
authors: van Berkel
authors: Erik A L
authors: EA
authors: Biessen
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D001057:Apolipoproteins E / Q000172:deficiency / Q000235:genetics; D019556:COS Cells; D019703:Calcineurin / Q000378:metabolism; D020212:Carotid Artery Injuries / Q000188:drug therapy* / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D017536:Carotid Artery, Common / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D016893:Carotid Stenosis / Q000188:drug therapy* / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D049109:Cell Proliferation / Q000187:drug effects*; D002522:Chlorocebus aethiops; D016572:Cyclosporine / Q000494:pharmacology; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D015195:Drug Design; D057809:HEK293 Cells; D006801:Humans; D006965:Hyperplasia; D007166:Immunosuppressive Agents / Q000737:chemistry / Q000494:pharmacology*; D019169:Jurkat Cells; D008213:Lymphocyte Activation / Q000187:drug effects*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008958:Models, Molecular; D015394:Molecular Structure; D016297:Mutagenesis, Site-Directed; D050778:NFATC Transcription Factors / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology*; D010538:Peritonitis / Q000188:drug therapy* / Q000276:immunology / Q000378:metabolism; D012008:Recurrence; D015398:Signal Transduction / Q000187:drug effects; D013329:Structure-Activity Relationship; D013601:T-Lymphocytes / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D014162:Transfection
chemical_list: D001057:Apolipoproteins E; D007166:Immunosuppressive Agents; D050778:NFATC Transcription Factors; D010455:Peptides; D016572:Cyclosporine; D019703:Calcineurin