Hypoxia-inducible vascular endothelial growth factor gene therapy using the oxygen-dependent degradation domain in myocardial ischemia

Autor: Hyun Ah Kim, Sung Wan Kim, Soyeon Lim, Sun Hwa Kim, Donghoon Choi, Ki-Chul Hwang, Hyung Ho Moon, Minhyung Lee
Rok vydání: 2010
Předmět:
Male
Vascular Endothelial Growth Factor A
Oxygen dependent degradation
medicine.medical_specialty
Myocardial ischemia
Genetic enhancement
Genetic Vectors
Myocardial Ischemia
Pharmaceutical Science
Vegf expression
Apoptosis
Enzyme-Linked Immunosorbent Assay
Simian virus 40
Transfection
Cell Line
Rats
Sprague-Dawley
chemistry.chemical_compound
Internal medicine
In Situ Nick-End Labeling
Medicine
Animals
Humans
Pharmacology (medical)
Hypoxia
Luciferases
Erythropoietin
Pharmacology
business.industry
Vascular disease
Protein Stability
Myocardium
Organic Chemistry
Genetic Therapy
Hypoxia (medical)
medicine.disease
Immunohistochemistry
Rats
Vascular endothelial growth factor
Oxygen
Disease Models
Animal
Endocrinology
chemistry
Cancer research
Molecular Medicine
medicine.symptom
Myocardial disease
business
Biotechnology
Plasmids
Zdroj: Pharmaceutical research. 27(10)
ISSN: 1573-904X
Popis: A hypoxia-inducible VEGF expression system with the oxygen-dependent degradation (ODD) domain was constructed and tested to be used in gene therapy for ischemic myocardial disease.Luciferase and VEGF expression vector systems were constructed with or without the ODD domain: pEpo-SV-Luc (or pEpo-SV-VEGF) and pEpo-SV-Luc-ODD (or pEpo-SV-VEGF-ODD). In vitro gene expression efficiency of each vector type was evaluated in HEK 293 cells under both hypoxic and normoxic conditions. The amount of VEGF protein was estimated by ELISA. The VEGF expression vectors with or without the ODD domain were injected into ischemic rat myocardium. Fibrosis, neovascularization, and cardiomyocyte apoptosis were assessed using Masson's trichrome staining, α-smooth muscle actin (α-SMA) immunostaining, and the TUNEL assay, respectively.The plasmid vectors containing ODD significantly improved the expression level of VEGF protein in hypoxic conditions. The enhancement of VEGF protein production was attributed to increased protein stability due to oxygen deficiency. In a rat model of myocardial ischemia, the pEpo-SV-VEGF-ODD group exhibited less myocardial fibrosis, higher microvessel density, and less cardiomyocyte apoptosis compared to the control groups (saline and pEpo-SV-VEGF treatments).An ODD-mediated VEGF expression system that facilitates VEGF-production under hypoxia may be useful in the treatment of ischemic heart disease.
Databáze: OpenAIRE
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