Combined transfer of human VEGF165 and HGF genes renders potent angiogenic effect in ischemic skeletal muscle

Autor: I. N. Rybalkin, I. B. Beloglazova, E.K. Shevchenko, Alexander Shevelev, T. N. Vlasik, Vsevolod A. Tkachuk, Yelena V. Parfyonova, Z. I. Tsokolaeva, Pavel I. Makarevich
Rok vydání: 2012
Předmět:
Male
Vascular Endothelial Growth Factor A
Anatomy and Physiology
Mouse
Angiogenesis
Genetic enhancement
Cardiovascular
Cardiovascular System
Biochemistry
Mice
Ischemia
Molecular Cell Biology
Membrane Receptor Signaling
Tube formation
Peripheral Vascular Diseases
Multidisciplinary
Neovascularization
Pathologic
Hepatocyte Growth Factor
Electroporation
Gene Therapy
Animal Models
Vascular endothelial growth factor A
medicine.anatomical_structure
Medicine
Human umbilical vein endothelial cell
Hepatocyte growth factor
medicine.drug
Research Article
Signal Transduction
Science
Blotting
Western
Enzyme-Linked Immunosorbent Assay
Endocrine System
Biology
Transfection
Model Organisms
Vascular Biology
Growth Factors
medicine
Human Umbilical Vein Endothelial Cells
Genetics
Animals
Humans
Muscle
Skeletal
Endocrine Physiology
Skeletal muscle
Proteins
Human Genetics
Molecular biology
Mice
Inbred C57BL
HEK293 Cells
Cancer research
Zdroj: PLoS ONE
PLoS ONE, Vol 7, Iss 6, p e38776 (2012)
ISSN: 1932-6203
Popis: Increased interest in development of combined gene therapy emerges from results of recent clinical trials that indicate good safety yet unexpected low efficacy of “single-gene” administration. Multiple studies showed that vascular endothelial growth factor 165 aminoacid form (VEGF165) and hepatocyte growth factor (HGF) can be used for induction of angiogenesis in ischemic myocardium and skeletal muscle. Gene transfer system composed of a novel cytomegalovirus-based (CMV) plasmid vector and codon-optimized human VEGF165 and HGF genes combined with intramuscular low-voltage electroporation was developed and tested in vitro and in vivo. Studies in HEK293T cell culture, murine skeletal muscle explants and ELISA of tissue homogenates showed efficacy of constructed plasmids. Functional activity of angiogenic proteins secreted by HEK293T after transfection by induction of tube formation in human umbilical vein endothelial cell (HUVEC) culture. HUVEC cells were used for in vitro experiments to assay the putative signaling pathways to be responsible for combined administration effect one of which could be the ERK1/2 pathway. In vivo tests of VEGF165 and HGF genes co-transfer were conceived in mouse model of hind limb ischemia. Intramuscular administration of plasmid encoding either VEGF165 or HGF gene resulted in increased perfusion compared to empty vector administration. Mice injected with a mixture of two plasmids (VEGF165+HGF) showed significant increase in perfusion compared to single plasmid injection. These findings were supported by increased CD31+ capillary and SMA+ vessel density in animals that received combined VEGF165 and HGF gene therapy compared to single gene therapy. Results of the study suggest that co-transfer of VEGF and HGF genes renders a robust angiogenic effect in ischemic skeletal muscle and may present interest as a potential therapeutic combination for treatment of ischemic disorders.
Databáze: OpenAIRE
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