Long-term cardiac gene expression using a coxsackieviral vector

Byung Kwan Lim, Jae Ok Shin, Sang Chol Lee, Duk Kyung Kim, Dong-Ju Choi, Seong Choon Choe, Kirk U. Knowlton, Eun Seok Jeon

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17 Citations (Scopus)

Abstract

Efficient myocardial gene transfer in the intact adult heart is difficult using conventional transfer vectors. Since coxsackievirus B3 (CVB3) is cardiotropic, it may be possible to exploit its cardiotropic characteristics to design a vector for gene transfer to the intact heart. We generated a recombinant CVB3 cDNA by inserting a green fluorescent protein (GFP) gene immediately upstream from the VP0 capsid protein of CVB3. The infectious virus (rCVB3-GFP) was recovered from the supernatants of the transfected Cos-7 cells, and was grown in HeLa cells to titers of 1011:pfu/ml. In the rCVB3-GFP infected HeLa cells and neonatal rat cardiac myocytes, GFP protein expression was documented by immunoblot and by fluorescent microscopy. GFP expression was maintained after five passages in HeLa cells. To test in vivo expression of GFP, we infected 8-week-old inbred female Balb/C mice with 10 6:pfu of rCVB3-GFP, intraperitoneally. GFP was present in up to 30% of cardiac myocytes over the 8:weeks post infection (p.i.) and it was co-localized with CVB3 infection. Surprisingly, in spite of detection of GFP up to at least 8:weeks after infection, there was no mortality in the mice. It is possible to express exogenous proteins in the intact heart after an intraperitoneal (i.p.) injection of recombinant coxsackievirus. The duration of expression persisted for at least 8:weeks with little immune response nor mortality. These results demonstrated that the cardiac tropism of CVB3 could be used to design vectors for efficient gene expression in the intact heart.

Original languageEnglish
Pages (from-to)745-751
Number of pages7
JournalJournal of Molecular and Cellular Cardiology
Volume38
Issue number5
DOIs
StatePublished - 1 Jan 2005

Fingerprint

Green Fluorescent Proteins
Gene Expression
Enterovirus
HeLa Cells
Cardiac Myocytes
Coxsackievirus Infections
Genes
Tropism
Mortality
Capsid Proteins
Infection
Intraperitoneal Injections
Microscopy
Proteins
Complementary DNA
Viruses

Keywords

  • Cardiac tropism
  • Coxsackievirus
  • Gene transfer
  • Myocarditis
  • Vector

Cite this

Lim, Byung Kwan ; Shin, Jae Ok ; Lee, Sang Chol ; Kim, Duk Kyung ; Choi, Dong-Ju ; Choe, Seong Choon ; Knowlton, Kirk U. ; Jeon, Eun Seok. / Long-term cardiac gene expression using a coxsackieviral vector. In: Journal of Molecular and Cellular Cardiology. 2005 ; Vol. 38, No. 5. pp. 745-751.
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abstract = "Efficient myocardial gene transfer in the intact adult heart is difficult using conventional transfer vectors. Since coxsackievirus B3 (CVB3) is cardiotropic, it may be possible to exploit its cardiotropic characteristics to design a vector for gene transfer to the intact heart. We generated a recombinant CVB3 cDNA by inserting a green fluorescent protein (GFP) gene immediately upstream from the VP0 capsid protein of CVB3. The infectious virus (rCVB3-GFP) was recovered from the supernatants of the transfected Cos-7 cells, and was grown in HeLa cells to titers of 1011:pfu/ml. In the rCVB3-GFP infected HeLa cells and neonatal rat cardiac myocytes, GFP protein expression was documented by immunoblot and by fluorescent microscopy. GFP expression was maintained after five passages in HeLa cells. To test in vivo expression of GFP, we infected 8-week-old inbred female Balb/C mice with 10 6:pfu of rCVB3-GFP, intraperitoneally. GFP was present in up to 30{\%} of cardiac myocytes over the 8:weeks post infection (p.i.) and it was co-localized with CVB3 infection. Surprisingly, in spite of detection of GFP up to at least 8:weeks after infection, there was no mortality in the mice. It is possible to express exogenous proteins in the intact heart after an intraperitoneal (i.p.) injection of recombinant coxsackievirus. The duration of expression persisted for at least 8:weeks with little immune response nor mortality. These results demonstrated that the cardiac tropism of CVB3 could be used to design vectors for efficient gene expression in the intact heart.",
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Lim, BK, Shin, JO, Lee, SC, Kim, DK, Choi, D-J, Choe, SC, Knowlton, KU & Jeon, ES 2005, 'Long-term cardiac gene expression using a coxsackieviral vector', Journal of Molecular and Cellular Cardiology, vol. 38, no. 5, pp. 745-751. https://doi.org/10.1016/j.yjmcc.2005.02.018

Long-term cardiac gene expression using a coxsackieviral vector. / Lim, Byung Kwan; Shin, Jae Ok; Lee, Sang Chol; Kim, Duk Kyung; Choi, Dong-Ju; Choe, Seong Choon; Knowlton, Kirk U.; Jeon, Eun Seok.

In: Journal of Molecular and Cellular Cardiology, Vol. 38, No. 5, 01.01.2005, p. 745-751.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Lim, Byung Kwan

AU - Shin, Jae Ok

AU - Lee, Sang Chol

AU - Kim, Duk Kyung

AU - Choi, Dong-Ju

AU - Choe, Seong Choon

AU - Knowlton, Kirk U.

AU - Jeon, Eun Seok

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KW - Cardiac tropism

KW - Coxsackievirus

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