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

✍ Scribed by Chester Bittencourt Sacramento; Vanessa Dionisio Cantagalli; Mariana Grings; Leonardo Pinto Carvalho; José Carlos Costa Baptista-Silva; Abram Beutel; Cassia Toledo Bergamaschi; Ruy Ribeiro de Campos Junior; Jane Zveiter de Moraes; Christina Maeda Takiya; Vívian Yochiko Samoto; Radovan Borojevic; Flavia Helena da Silva; Nance Beyer Nardi; Hans Fernando Dohmann; Hamilton Silva Junior; Valderez Bastos Valero; Sang Won Han

• Publisher: John Wiley and Sons
• Year: 2009
• Tongue: English
• Weight: 334 KB
• Volume: 11
• Category: Article
• ISSN: 1099-498X
• DOI: 10.1002/jgm.1298

✦ Synopsis

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 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.