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Hypoxia-Inducible Vascular Endothelial Growth Factor Gene Therapy Using the Oxygen-Dependent Degradation Domain in Myocardial Ischemia

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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.

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ACKNOWLEDGMENTS

This research was supported by grants from the Korea Healthcare Technology R&D Project, Ministry for Health, Welfare & Family Affairs, Republic of Korea (A085136); a faculty research grant of Yonsei University College of Medicine for 2009 (8-2009-0187); and the National Research Foundation of Korea (NRF), funded by the Ministry of Education, Science and Technology (20090065404 and 20090081874).

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Authors and Affiliations

  1. Department of Bioengineering, College of Engineering, Hanyang University, Seoul, 133-791, Korea

    Hyun Ah Kim, Sung Wan Kim & Minhyung Lee

  2. Severance Integrative Research Institute for Cerebral and Cardiovascular Disease, Yonsei University Health System, 250 Seongsanno, Seodaemun-gu, Seoul, 120-752, South Korea

    Soyeon Lim, Hyung-Ho Moon, Ki-Chul Hwang, Sun Hwa Kim & Donghoon Choi

  3. Center for Controlled Chemical Delivery, Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, Salt Lake City, Utah, 84112, USA

    Sung Wan Kim

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  1. Hyun Ah Kim
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  2. Soyeon Lim
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  3. Hyung-Ho Moon
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  5. Ki-Chul Hwang
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  7. Sun Hwa Kim
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Correspondence to Sun Hwa Kim or Donghoon Choi.

Additional information

Hyun Ah Kim and Soyeon Lim contributed equally to this work.

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Kim, H.A., Lim, S., Moon, HH. et al. Hypoxia-Inducible Vascular Endothelial Growth Factor Gene Therapy Using the Oxygen-Dependent Degradation Domain in Myocardial Ischemia. Pharm Res 27, 2075–2084 (2010). https://doi.org/10.1007/s11095-010-0206-7

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  • DOI: https://doi.org/10.1007/s11095-010-0206-7

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