title: Granulocyte-macrophage colony-stimulating factor gene based therapy for acute limb ischemia in a mouse model.
abstract: BACKGROUND
  Granulocyte-colony-stimulating factor (GM-CSF) is a pleiotropic factor for hematopoiesis that stimulates myeloblasts, monoblasts and mobilization of bone marrow stem cells. Therefore, the GM-CSF gene is a potential candidate for vessel formation and tissue remodeling in the treatment of ischemic diseases.
  
  
  METHODS
  A new mouse limb ischemia was established by surgery and gene transfer was performed by injection of 100 microg of a plasmid carrying GM-CSF. Muscle force and weight, histology, capillary density, circulating stem cells and monocytes were determined after 3-4 weeks.
  
  
  RESULTS
  More than 60% of nontreated ischemic animals showed gangrene below the heel after 4 weeks, whereas the GM-CSF gene-treated animals showed only darkening of nails or toes. These animals demonstrated a full recovery of the affected muscles in terms of weight, force and muscle fiber structure, but the muscles of nontreated ischemic animals lost approximately 50% weight, 86% force and their regular structure. When the GM-CSF gene was injected into the contralateral limb, only partial loss was observed, demonstrating a distant effect of GM-CSF. The capillary density in the GM-CSF-treated group was 52% higher in relation to the nontreated group. Blood analysis by flow cytometry showed that the GM-CSF-treated group had 10-20% higher levels of circulating monocytes and Sca-1(+).
  
  
  CONCLUSIONS
  We conclude that the direct administration of GM-CSF gene in limb ischemia had a strong therapeutic effect because it promoted the recovery of muscle mass, force and structure by mobilizing therapeutic cells and augmenting the number of vessels.
authors.affiliation: Interdisciplinary Center for Gene Therapy, Federal University of São Paulo, São Paulo, Brazil.
authors.forename: Chester Bittencourt
authors.initials: CB
authors.lastname: Sacramento
authors.forename: Vanessa Dionisio
authors.initials: VD
authors.lastname: Cantagalli
authors.forename: Mariana
authors.initials: M
authors.lastname: Grings
authors.forename: Leonardo Pinto
authors.initials: LP
authors.lastname: Carvalho
authors.forename: José Carlos Costa
authors.initials: JC
authors.lastname: Baptista-Silva
authors.forename: Abram
authors.initials: A
authors.lastname: Beutel
authors.forename: Cassia Toledo
authors.initials: CT
authors.lastname: Bergamaschi
authors.forename: Ruy Ribeiro
authors.initials: RR
authors.lastname: de Campos Junior
authors.forename: Jane Zveiter
authors.initials: JZ
authors.lastname: de Moraes
authors.forename: Christina Maeda
authors.initials: CM
authors.lastname: Takiya
authors.forename: Vívian Yochiko
authors.initials: VY
authors.lastname: Samoto
authors.forename: Radovan
authors.initials: R
authors.lastname: Borojevic
authors.forename: Flavia Helena
authors.initials: FH
authors.lastname: da Silva
authors.forename: Nance Beyer
authors.initials: NB
authors.lastname: Nardi
authors.forename: Hans Fernando
authors.initials: HF
authors.lastname: Dohmann
authors.forename: Hamilton Silva
authors.initials: HS
authors.lastname: Junior
authors.forename: Valderez Bastos
authors.initials: VB
authors.lastname: Valero
authors.forename: Sang Won
authors.initials: SW
authors.lastname: Han
mesh_terms: D000208:Acute Disease; D000818:Animals; D004195:Disease Models, Animal; D005121:Extremities / Q000473:pathology; D015316:Genetic Therapy / Q000379:methods*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000008:administration & dosage*; D006410:Hematopoiesis / Q000187:drug effects; D007511:Ischemia / Q000628:therapy*; D051379:Mice; D018482:Muscle, Skeletal / Q000187:drug effects; D018919:Neovascularization, Physiologic / Q000187:drug effects; D010957:Plasmids / Q000008:administration & dosage; D016896:Treatment Outcome
chemical_list: D016178:Granulocyte-Macrophage Colony-Stimulating Factor