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

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...

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Autores principales: Pavel Makarevich, Zoya Tsokolaeva, Alexander Shevelev, Igor Rybalkin, Evgeny Shevchenko, Irina Beloglazova, Tatyana Vlasik, Vsevolod Tkachuk, Yelena Parfyonova
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Publicado: Public Library of Science (PLoS) 2012
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spelling oai:doaj.org-article:ccb99f37a0ce4bbca79c397bebfc1c9c2021-11-18T07:15:37ZCombined transfer of human VEGF165 and HGF genes renders potent angiogenic effect in ischemic skeletal muscle.1932-620310.1371/journal.pone.0038776https://doaj.org/article/ccb99f37a0ce4bbca79c397bebfc1c9c2012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22719942/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Increased 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.Pavel MakarevichZoya TsokolaevaAlexander ShevelevIgor RybalkinEvgeny ShevchenkoIrina BeloglazovaTatyana VlasikVsevolod TkachukYelena ParfyonovaPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 6, p e38776 (2012)
institution DOAJ
collection DOAJ
language EN
topic Medicine
R
Science
Q
spellingShingle Medicine
R
Science
Q
Pavel Makarevich
Zoya Tsokolaeva
Alexander Shevelev
Igor Rybalkin
Evgeny Shevchenko
Irina Beloglazova
Tatyana Vlasik
Vsevolod Tkachuk
Yelena Parfyonova
Combined transfer of human VEGF165 and HGF genes renders potent angiogenic effect in ischemic skeletal muscle.
description 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.
format article
author Pavel Makarevich
Zoya Tsokolaeva
Alexander Shevelev
Igor Rybalkin
Evgeny Shevchenko
Irina Beloglazova
Tatyana Vlasik
Vsevolod Tkachuk
Yelena Parfyonova
author_facet Pavel Makarevich
Zoya Tsokolaeva
Alexander Shevelev
Igor Rybalkin
Evgeny Shevchenko
Irina Beloglazova
Tatyana Vlasik
Vsevolod Tkachuk
Yelena Parfyonova
author_sort Pavel Makarevich
title Combined transfer of human VEGF165 and HGF genes renders potent angiogenic effect in ischemic skeletal muscle.
title_short Combined transfer of human VEGF165 and HGF genes renders potent angiogenic effect in ischemic skeletal muscle.
title_full Combined transfer of human VEGF165 and HGF genes renders potent angiogenic effect in ischemic skeletal muscle.
title_fullStr Combined transfer of human VEGF165 and HGF genes renders potent angiogenic effect in ischemic skeletal muscle.
title_full_unstemmed Combined transfer of human VEGF165 and HGF genes renders potent angiogenic effect in ischemic skeletal muscle.
title_sort combined transfer of human vegf165 and hgf genes renders potent angiogenic effect in ischemic skeletal muscle.
publisher Public Library of Science (PLoS)
publishDate 2012
url https://doaj.org/article/ccb99f37a0ce4bbca79c397bebfc1c9c
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