title: Activators and stimulators of soluble guanylate cyclase counteract myofibroblast differentiation of prostatic and dermal stromal cells.
abstract: BACKGROUND
  Fibrotic diseases encompass numerous systemic and organ-specific disorders characterized by the development and persistence of myofibroblasts. TGFβ1 is considered the key inducer of fibrosis and drives myofibroblast differentiation in cells of diverse histological origin by a pro-oxidant shift in redox homeostasis associated with decreased nitric oxide (NO)/cGMP signaling. Thus, enhancement of NO/cGMP represents a potential therapeutic strategy to target myofibroblast activation and therefore fibrosis.
  
  
  METHODS
  Myofibroblast differentiation was induced by TGFβ1 in human primary prostatic (PrSCs) and normal dermal stromal cells (NDSCs) and monitored by α smooth muscle cell actin (SMA) and IGF binding protein 3 (IGFBP3) mRNA and protein levels. The potential of enhanced cGMP production by the sGC stimulator BAY 41-2272 or the sGC activator BAY 60-2770 to inhibit and revert myofibroblast differentiation in vitro was analyzed. Moreover, potential synergisms of BAY 41-2272 or BAY 60-2770 and inhibition of cGMP degradation by the PDE5 inhibitor vardenafil were investigated.
  
  
  RESULTS
  BAY 41-2272 and BAY 60-2770 at doses of 30µM significantly inhibited induction of SMA and IGFBP3 levels in PrSCs and reduced myofibroblast marker levels in TGFβ1-predifferentiated cells. At lower concentrations (3 and 10µM) only BAY 41-2272 but not BAY 60-2770 significantly inhibited and reverted myofibroblast differentiation. In NDSCs both substances significantly inhibited differentiation at all concentrations tested. Attenuation of SMA expression was more pronounced in NDSCs whereas reduction of IGFBP3 levels by BAY 41-2272 appeared more efficient in PrSCs. Moreover, administration of BAY 41-2272 or BAY 60-2770 enhanced the efficiency of the PDE5 inhibitor vardenafil to inhibit and revert myofibroblast differentiation in vitro.
  
  
  CONCLUSIONS
  Increase of cGMP by sGC stimulation/activation significantly inhibited and reverted myofibroblast differentiation. This effect was even more pronounced when a combination treatment with a PDE5 inhibitor was applied. Thus, enhancement of NO/cGMP-signaling by sGC stimulation/activation is a promising strategy for the treatment of fibrotic diseases. Whereas, in NDSCs BAY 60-2770 and BAY 41-2272 exerted similar effects on myofibroblast differentiation, higher potency of BAY 41-2272 was observed in PrSCs, indicating phenotypical differences between fibroblasts form different organs that should be taken into account in the search for antifibrotic therapies.
authors.affiliation: University of Applied Sciences Tyrol, Innsbruck, Austria. Electronic address: christoph.zenzmaier@fhg-tirol.ac.at.
authors.forename: Christoph
authors.initials: C
authors.lastname: Zenzmaier
authors.affiliation: Oncotyrol Laboratory for Tumor Biology and Angiogenesis, Innsbruck, Austria. Electronic address: Johann.Kern@oncotyrol.at.
authors.forename: Johann
authors.initials: J
authors.lastname: Kern
authors.affiliation: Institute for Biomedical Aging Research, University of Innsbruck, Innsbruck, Austria. Electronic address: Martin.Heitz@uibk.ac.at.
authors.forename: Martin
authors.initials: M
authors.lastname: Heitz
authors.affiliation: Department of Urology, Hanusch Hospital, Vienna, Austria. Electronic address: eugen.plas@wgkk.at.
authors.forename: Eugen
authors.initials: E
authors.lastname: Plas
authors.affiliation: Institute for Biomedical Aging Research, University of Innsbruck, Innsbruck, Austria. Electronic address: Werner.Zwerschke@uibk.ac.at.
authors.forename: Werner
authors.initials: W
authors.lastname: Zwerschke
authors.affiliation: Department of Plastic and Reconstructive Surgery, Innsbruck Medical University, Innsbruck, Austria. Electronic address: monika.mattesich@i-med.ac.at.
authors.forename: Monika
authors.initials: M
authors.lastname: Mattesich
authors.affiliation: Bayer HealthCare, Global Drug Discovery, Heart and Lung Diseases, Wuppertal, Germany and Hannover Medical School, Department of Pharmacology, Hannover, Germany. Electronic address: peter.sandner@bayer.com.
authors.forename: Peter
authors.initials: P
authors.lastname: Sandner
authors.affiliation: Institute for Biomedical Aging Research, University of Innsbruck, Innsbruck, Austria. Electronic address: Peter.Berger@uibk.ac.at.
authors.forename: Peter
authors.initials: P
authors.lastname: Berger
mesh_terms: D000199:Actins / Q000378:metabolism; D001565:Benzoates / Q000494:pharmacology; D001713:Biphenyl Compounds / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005355:Fibrosis / Q000378:metabolism; D006162:Guanylate Cyclase / Q000378:metabolism*; D006801:Humans; D006845:Hydrocarbons, Fluorinated / Q000494:pharmacology; D018972:Insulin-Like Growth Factor Binding Protein 3 / Q000378:metabolism; D008297:Male; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000378:metabolism; D058628:Myofibroblasts / Q000187:drug effects / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism; D011467:Prostate / Q000187:drug effects / Q000378:metabolism*; D011720:Pyrazoles / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D000071756:Soluble Guanylyl Cyclase; D017154:Stromal Cells / Q000187:drug effects / Q000378:metabolism*; D016212:Transforming Growth Factor beta / Q000378:metabolism; D000099222:betaIG-H3 Protein
chemical_list: D000199:Actins; D001565:Benzoates; D001713:Biphenyl Compounds; D006152:Cyclic GMP; D016326:Extracellular Matrix Proteins; D006162:Guanylate Cyclase; D006845:Hydrocarbons, Fluorinated; D018972:Insulin-Like Growth Factor Binding Protein 3; D009569:Nitric Oxide; D011720:Pyrazoles; D011725:Pyridines; D018160:Receptors, Cytoplasmic and Nuclear; D000071756:Soluble Guanylyl Cyclase; D016212:Transforming Growth Factor beta; D000099222:betaIG-H3 Protein; C423582:3-(4-Amino-5-cyclopropylpyrimidine-2-yl)-1-(2-fluorobenzyl)-1H-pyrazolo(3,4-b)pyridine; C528797:4-(((4-carboxybutyl) (2- (5-fluoro-2-((4'-(trifluoromethyl) biphenyl-4-yl)methoxy)phenyl)ethyl) amino)methyl)benzoic acid; C515497:IGFBP3 protein, human
keywords: Fibrosis; Myofibroblast; NO/cGMP signaling; PDE5 inhibition; sGC activation; sGC stimulation