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

Hyun Ah Kim, Soyeon Lim, Hyung Ho Moon, Sung Wan Kim, Ki Chul Hwang, Minhyung Lee, Sun Hwa Kim, Donghoon Choi

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Purpose: 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. Methods: 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. Results: 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). Conclusion: An ODD-mediated VEGF expression system that facilitates VEGF-production under hypoxia may be useful in the treatment of ischemic heart disease.

Original languageEnglish
Pages (from-to)2075-2084
Number of pages10
JournalPharmaceutical Research
Volume27
Issue number10
DOIs
Publication statusPublished - 2010 Oct 1

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Gene therapy
Genetic Therapy
Vascular Endothelial Growth Factor A
Myocardial Ischemia
Oxygen
Degradation
Cardiac Myocytes
Hypoxia
Rats
Proteins
Fibrosis
Apoptosis
Protein Stability
HEK293 Cells
In Situ Nick-End Labeling
Microvessels
Luciferases
Cardiomyopathies
Gene expression
Smooth Muscle

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Molecular Medicine
  • Pharmacology
  • Pharmaceutical Science
  • Organic Chemistry
  • Pharmacology (medical)

Cite this

Kim, Hyun Ah ; Lim, Soyeon ; Moon, Hyung Ho ; Kim, Sung Wan ; Hwang, Ki Chul ; Lee, Minhyung ; Kim, Sun Hwa ; Choi, Donghoon. / Hypoxia-inducible vascular endothelial growth factor gene therapy using the oxygen-dependent degradation domain in myocardial ischemia. In: Pharmaceutical Research. 2010 ; Vol. 27, No. 10. pp. 2075-2084.
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Hypoxia-inducible vascular endothelial growth factor gene therapy using the oxygen-dependent degradation domain in myocardial ischemia. / Kim, Hyun Ah; Lim, Soyeon; Moon, Hyung Ho; Kim, Sung Wan; Hwang, Ki Chul; Lee, Minhyung; Kim, Sun Hwa; Choi, Donghoon.

In: Pharmaceutical Research, Vol. 27, No. 10, 01.10.2010, p. 2075-2084.

Research output: Contribution to journalArticle

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AU - Kim, Hyun Ah

AU - Lim, Soyeon

AU - Moon, Hyung Ho

AU - Kim, Sung Wan

AU - Hwang, Ki Chul

AU - Lee, Minhyung

AU - Kim, Sun Hwa

AU - Choi, Donghoon

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N2 - Purpose: 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. Methods: 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. Results: 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). Conclusion: An ODD-mediated VEGF expression system that facilitates VEGF-production under hypoxia may be useful in the treatment of ischemic heart disease.

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