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

Author(s):Show less -

Sacramento, Chester Bittencourt ^[1] ; Cantagalli, Vanessa Dionisio ^[1] ; Grings, Mariana ^[1] ; Carvalho, Leonardo Pinto ^[1] ; Costa Baptista-Silva, Jose Carlos ^[2] ; Beutel, Abram ^[3] ; Bergamaschi, Cassia Toledo ^[3] ; de Campos Junior, Ruy Ribeiro ^[3] ; de Moraes, Jane Zveiter ^[4] ; Takiya, Christina Maeda ^[5] ; Samoto, Vivian Yochiko ^[5] ; Borojevic, Radovan ^[5] ; da Silva, Flavia Helena ^{[1, 6]} ; Nardi, Nance Beyer ^[6] ; Dohmann, Hans Fernando ^[7] ; Silva Junior, Hamilton ^[7] ; Valero, Valderez Bastos ^[1] ; Han, Sang Won ^{[4, 1]}

Affiliation

^[1] Univ Fed Sao Paulo, Interdisciplinary Ctr Gene Therapy, Sao Paulo - Brazil

^[2] Univ Fed Sao Paulo, Dept Surg, Sao Paulo - Brazil

^[3] Univ Fed Sao Paulo, Dept Physiol, Sao Paulo - Brazil

^[4] Univ Fed Sao Paulo, Dept Biophys, Sao Paulo - Brazil

^[5] Univ Fed Rio de Janeiro, Dept Histol & Embryol, Rio De Janeiro - Brazil

^[6] Univ Fed Rio Grande do Sul, Biosci Inst, Dept Genet, Rio Grande Do Sul - Brazil

^[7] Excell Biomed Serv, Petropolis, RJ - Brazil

• Document type: Journal article
• Source: JOURNAL OF GENE MEDICINE; v. 11, n. 4, p. 345-353, APR 2009.
• Web of Science Citations: 11

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 mu g 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 GMCSF-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 (C) 2009 John Wiley \& Sons, Ltd. (AU)