Synergistic effect of vascular endothelial growth factor and granulocyte colony-stimulating factor double gene therapy in mouse limb ischemia.
BACKGROUND
Vascular endothelial growth factor (VEGF) has mostly been tested to treat ischemic diseases, although the outcomes obtained are not satisfactory. Our hypothesis is that the local transient expression of VEGF and stem cell mobilizer granulocyte colony-stimulating factor (G-CSF) genes in ischemic limbs can complement their activities and be more efficient for limb recovery.


METHODS
Limb ischemia was surgically induced in mice and 50 microg of VEGF and/or G-CSF genes were locally transferred by electroporation. After 3-4 weeks, evidence of necrosis by visual inspection, capillary density, muscle mass, muscle force and hematopoietic cell mobilization were evaluated.


RESULTS
After 4 weeks, 70% and 90% of the animals of the ischemic group (IG) and VEGF-treated group (VG), respectively, presented limb necrosis, in contrast to only 10% observed in the group of mice treated with both VEGF and G-CSF genes (VGG). Recovery of muscle mass and muscle force was higher than 60% in the VGG compared to the non-ischemic group. The mobilization of Sca1+ cells and neutrophils was also higher in the VGG, which may explain the lower level of necrosis observed in this group (22%, in contrast to 70% in the IG). Capillary density and degree of fibrosis were determined in weeks 3 and 4, and also showed a clear benefit as a result of the use of the G-CSF and VEGF genes together.


CONCLUSIONS
Gene therapy using VEGF and G-CSF demonstrated a synergistic effect promoting vessel and tissue repair in mouse hind limb ischemia.
Sacramento
Chester Bittencourt
CB
Interdisciplinary center for gene therapy, Federal University of São Paulo, São Paulo-SP, 04044-010, Brazil.
da Silva
Flavia Helena
FH
Nardi
Nance Beyer
NB
Yasumura
Eduardo Gallatti
EG
Baptista-Silva
José Carlos Costa
JC
Beutel
Abram
A
de Campos
Ruy Ribeiro
RR
de Moraes
Jane Zveiter
JZ
Junior
Hamilton Silva
HS
Samoto
Vivian Yochiko
VY
Borojevic
Radovan
R
Han
Sang Won
SW
D000818:Animals; D005121:Extremities / Q000098:blood supply*; D015316:Genetic Therapy / Q000379:methods*; D016179:Granulocyte Colony-Stimulating Factor / Q000235:genetics*; D007511:Ischemia / Q000097:blood / Q000209:etiology / Q000628:therapy*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D018482:Muscle, Skeletal / Q000098:blood supply / Q000502:physiology; D018919:Neovascularization, Physiologic / Q000235:genetics; D016491:Peripheral Vascular Diseases / Q000150:complications / Q000628:therapy*; D012038:Regeneration / Q000235:genetics; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics*
D042461:Vascular Endothelial Growth Factor A; D016179:Granulocyte Colony-Stimulating Factor