Restoration of blood flow by using continuous perimuscular infiltration of plasmid DNA encoding subterranean mole rat Spalax ehrenbergi VEGF.
The optimal vector, regulatory sequences, and method of delivery of angiogenic gene therapy are of considerable interest. The Spalax ehrenbergi superspecies live in subterranean burrows at low oxygen tensions and its tissues are highly vascularized. We tested whether continuous perimuscular administration of Spalax vascular endothelial growth factor (VEGF) DNA could increase tissue perfusion in a murine hindlimb ischemia model. Placebo or VEGF +/- internal ribosome entry site (IRES) was continuously administrated perimuscularly in the ischemic zone by using an infusion pump. None of the mice in the VEGF-treated group (>50 microg) developed visible necrosis vs. 33% of the placebo group. Microscopic necrosis was observed only in the placebo group. Spalax VEGF muscular infiltration resulted in a faster and more complete restoration of blood flow. The restoration of blood flow by VEGF was dose-dependent and more robust and rapid when using the VEGF-IRES elements. The flow restoration using continuous perimuscular infiltration was faster than single i.m. injections. Vessel density was higher in the VEGF and VEGF-IRES (-) groups compared with the placebo. Continuous perimuscular administration of angiogenic gene therapy offers a new approach to restore blood flow to an ischemic limb. Incorporation of an IRES element may assist in the expression of transgenes delivered to ischemic tissues. Further studies are needed to determine whether VEGF from the subterranean mole rat Spalax VEGF is superior to VEGF from other species. If so, 40 million years of Spalax evolution underground, including adaptive hypoxia tolerance, may prove important to human angiogenic gene therapy.
Roguin
Ariel
A
Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, POB 9649, Haifa 31096, Israel.
Avivi
Aaron
A
Nitecki
Samy
S
Rubinstein
Irit
I
Levy
Nina S
NS
Abassi
Zaid A
ZA
Resnick
Murray B
MB
Lache
Orit
O
Melamed-Frank
Meira
M
Joel
Alma
A
Hoffman
Aaron
A
Nevo
Eviatar
E
Levy
Andrew P
AP
D000818:Animals; D001783:Blood Flow Velocity / Q000187:drug effects; D016228:Endothelial Growth Factors / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D015316:Genetic Therapy; D006614:Hindlimb / Q000098:blood supply; D015918:Infusion Pumps, Implantable; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D007511:Ischemia / Q000503:physiopathology / Q000628:therapy*; D008222:Lymphokines / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D019577:Mole Rats / Q000235:genetics*; D018919:Neovascularization, Physiologic / Q000187:drug effects / Q000235:genetics*; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D016228:Endothelial Growth Factors; D036341:Intercellular Signaling Peptides and Proteins; D008222:Lymphokines; D011994:Recombinant Proteins; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors