Granulocyte-macrophage colony-stimulating factor gene based therapy for acute limb ischemia in a mouse model

• Published in: The journal of gene medicine 2009-04, Vol.11 (4), p.345-353
• Main Authors: Sacramento, Chester Bittencourt, Cantagalli, Vanessa Dionisio, Grings, Mariana, Carvalho, Leonardo Pinto, Baptista-Silva, José Carlos Costa, Beutel, Abram, Bergamaschi, Cassia Toledo, de Campos Junior, Ruy Ribeiro, de Moraes, Jane Zveiter, Takiya, Christina Maeda, Samoto, Vívian Yochiko, Borojevic, Radovan, da Silva, Flavia Helena, Nardi, Nance Beyer, Dohmann, Hans Fernando, Junior, Hamilton Silva, Valero, Valderez Bastos, Han, Sang Won
• Format: Article
• Language: English
• Subjects: Acute Disease; angiogenesis; Animals; arteriogenesis; Disease Models, Animal; Extremities - pathology; Gene therapy; Genetic Therapy - methods; GM-CSF; Granulocyte-Macrophage Colony-Stimulating Factor - administration & dosage; Hematopoiesis - drug effects; ischemia; Ischemia - therapy; Mice; Muscle, Skeletal - drug effects; Neovascularization, Physiologic - drug effects; Plasmids - administration & dosage; Treatment Outcome; vasculogenesis
• ISSN: 1099-498X; 1521-2254
• DOI: 10.1002/jgm.1298

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 µg 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. Copyright © 2009 John Wiley & Sons, Ltd.