title: Fasudil, a clinically safe ROCK inhibitor, decreases disease burden in a Cbl/Cbl-b deficiency-driven murine model of myeloproliferative disorders.
abstract: OBJECTIVE
  Mutations in Cbl or Cbl-b gene occur in 10% of myeloproliferative disorder (MPD) patients and are associated with poor prognosis. Hematopoietic Cbl/Cbl-b double knockout (DKO) leads to a disease in mice phenotypically similar to human MPDs. The aim of this study was to evaluate the anti-MPD activity of a clinically safe drug, Fasudil, identified in an in vitro kinase inhibitor as an inhibitor of proliferation of DKO mouse hematopoietic stem/progenitor cells (HSPCs).
  
  
  METHODS
  Fasudil exhibited relatively selective anti-proliferative activity against Cbl/Cbl-b DKO vs. control murine bone marrow HSPCs. We established a mouse model with uniform time of MPD onset by transplanting Cbl/Cbl-b DKO HSPCs into busulfan-conditioned NOD/SCID/gamma chain-deficient mice. Four weeks post-transplant, mice were treated with 100 mg/kg fasudil (13 mice) or water (control, 8 mice) daily by oral gavage, followed by blood cell count every 2 weeks.
  
  
  RESULTS
  By 2 weeks of treatment, total white cell and monocyte counts were significantly lower in mice treated with fasudil. We observed a trend towards improved survival in fasudil-treated mice that did not reach statistical significance. Notably, prolonged survival beyond 27 weeks was observed in two fasudil-treated mice, nearly twice the 16-week average life-span in the Cbl/Cbl-b DKO MPD model.
  
  
  CONCLUSIONS
  Our results suggest a therapeutic potential for fasudil, a clinically safe drug with promising results in vascular diseases, in the treatment of MPDs or other mutant Cbl-driven myeloid disorders.
authors.affiliation: a Division of Hematology and Oncology, Department of Medicine , University of Nebraska Medical Center , Omaha , USA.
authors.forename: Basem M
authors.initials: BM
authors.lastname: William
authors.affiliation: b Department of Genetics, Cell Biology and Anatomy , University of Nebraska Medical Center , Omaha , USA.|c Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center , Omaha , USA.
authors.forename: Wei
authors.initials: W
authors.lastname: An
authors.affiliation: c Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center , Omaha , USA.
authors.forename: Dan
authors.initials: D
authors.lastname: Feng
authors.affiliation: b Department of Genetics, Cell Biology and Anatomy , University of Nebraska Medical Center , Omaha , USA.|c Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center , Omaha , USA.
authors.forename: Scott
authors.initials: S
authors.lastname: Nadeau
authors.affiliation: c Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center , Omaha , USA.|d Department of Biochemistry and Molecular Biology , University of Nebraska Medical Center , Omaha , USA.
authors.forename: Bhopal C
authors.initials: BC
authors.lastname: Mohapatra
authors.affiliation: c Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center , Omaha , USA.
authors.forename: Matthew A
authors.initials: MA
authors.lastname: Storck
authors.affiliation: b Department of Genetics, Cell Biology and Anatomy , University of Nebraska Medical Center , Omaha , USA.|c Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center , Omaha , USA.|e Fred & Pamela Buffett Cancer Center, University of Nebraska Medical Center , Omaha , USA.
authors.forename: Vimla
authors.initials: V
authors.lastname: Band
authors.affiliation: b Department of Genetics, Cell Biology and Anatomy , University of Nebraska Medical Center , Omaha , USA.|c Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center , Omaha , USA.|d Department of Biochemistry and Molecular Biology , University of Nebraska Medical Center , Omaha , USA.|e Fred & Pamela Buffett Cancer Center, University of Nebraska Medical Center , Omaha , USA.
authors.forename: Hamid
authors.initials: H
authors.lastname: Band
mesh_terms: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives* / Q000494:pharmacology; D048868:Adaptor Proteins, Signal Transducing / Q000172:deficiency*; D000818:Animals; D004195:Disease Models, Animal; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D018345:Mice, Knockout; D016513:Mice, SCID; D009196:Myeloproliferative Disorders / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism; D050721:Proto-Oncogene Proteins c-cbl / Q000172:deficiency*; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C118387:Cblb protein, mouse; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D050721:Proto-Oncogene Proteins c-cbl; D054460:rho-Associated Kinases; C493849:Cbl protein, mouse; C049347:fasudil
keywords: Cbl; Fasudil; Mouse model; Myeloproliferative diseases; Myosin light chain; Rho kinase