Quantitative determination of adenovirus-mediated gene delivery to rat cardiac myocytes in vitro and in vivo

Alyson Kass-Eisler, Erik Falck-Pedersen, Mauricio Alvira, Johanna Rivera, Peter M. Buttrick, Beatrice A. Wittenberg, Laura Cipriani, Leslie A. Leinwand

Research output: Contribution to journalArticle

216 Citations (Scopus)

Abstract

To optimize the use of modified adenoviruses as vectors for gene delivery to the myocardium, we have characterized infection of cultured fetal and adult rat cardiac myocytes in vitro and of adult cardiac myocytes in vivo by using a replication-defective adenovirus carrying the chloramphenicol acetyltransferase (CAT) reporter gene driven by the cytomegalovirus promoter (AdCMVCATgD). In vitro, virtually all fetal or adult cardiocytes express the CAT gene when infected with 1 plaque-forming unit of virus per cell. CAT enzymatic activity can be detected in these cells as early as 4 hr after infection, reaching near-maximal levels at 48 hr. In fetal cells, CAT expression was maintained without a loss in activity for at least 1 week. Using in vitro studies as a guide, we introduced the AdCMVCATgD virus directly into adult rat myocardium and compared the expression results obtained from virus injection with those obtained by direct injection of pAdCMVCATgD plasmid DNA. The amount of CAT activity resulting from adenovirus infection of the myocardium was orders of magnitude higher than that seen from DNA injection and was proportional to the amount of input virus. Immunostaining for CAT protein in cardiac tissue sections following adenovirus injection demonstrated large numbers of positive cells, reaching nearly 100% of the myocytes in many regions of the heart. Expression of genes introduced by adenovirus peaked at 5 days but was still detectable 55 days following infection. Adenoviruses are therefore a very useful tool for high-efficiency gene transfer into the cardiovascular system.

Original languageEnglish (US)
Pages (from-to)11498-11502
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume90
Issue number24
StatePublished - Dec 15 1993

Fingerprint

Chloramphenicol O-Acetyltransferase
Cardiac Myocytes
Adenoviridae
Genes
Viruses
Myocardium
Injections
Infection
Adenoviridae Infections
DNA
Cardiovascular System
Cytomegalovirus
Reporter Genes
Muscle Cells
In Vitro Techniques
Plasmids
Cell Count
Gene Expression
Proteins

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

Kass-Eisler, A., Falck-Pedersen, E., Alvira, M., Rivera, J., Buttrick, P. M., Wittenberg, B. A., ... Leinwand, L. A. (1993). Quantitative determination of adenovirus-mediated gene delivery to rat cardiac myocytes in vitro and in vivo. Proceedings of the National Academy of Sciences of the United States of America, 90(24), 11498-11502.

Quantitative determination of adenovirus-mediated gene delivery to rat cardiac myocytes in vitro and in vivo. / Kass-Eisler, Alyson; Falck-Pedersen, Erik; Alvira, Mauricio; Rivera, Johanna; Buttrick, Peter M.; Wittenberg, Beatrice A.; Cipriani, Laura; Leinwand, Leslie A.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 90, No. 24, 15.12.1993, p. 11498-11502.

Research output: Contribution to journalArticle

Kass-Eisler, A, Falck-Pedersen, E, Alvira, M, Rivera, J, Buttrick, PM, Wittenberg, BA, Cipriani, L & Leinwand, LA 1993, 'Quantitative determination of adenovirus-mediated gene delivery to rat cardiac myocytes in vitro and in vivo', Proceedings of the National Academy of Sciences of the United States of America, vol. 90, no. 24, pp. 11498-11502.
Kass-Eisler, Alyson ; Falck-Pedersen, Erik ; Alvira, Mauricio ; Rivera, Johanna ; Buttrick, Peter M. ; Wittenberg, Beatrice A. ; Cipriani, Laura ; Leinwand, Leslie A. / Quantitative determination of adenovirus-mediated gene delivery to rat cardiac myocytes in vitro and in vivo. In: Proceedings of the National Academy of Sciences of the United States of America. 1993 ; Vol. 90, No. 24. pp. 11498-11502.
@article{9a1c75d6df40450b922ebaef788c8c92,
title = "Quantitative determination of adenovirus-mediated gene delivery to rat cardiac myocytes in vitro and in vivo",
abstract = "To optimize the use of modified adenoviruses as vectors for gene delivery to the myocardium, we have characterized infection of cultured fetal and adult rat cardiac myocytes in vitro and of adult cardiac myocytes in vivo by using a replication-defective adenovirus carrying the chloramphenicol acetyltransferase (CAT) reporter gene driven by the cytomegalovirus promoter (AdCMVCATgD). In vitro, virtually all fetal or adult cardiocytes express the CAT gene when infected with 1 plaque-forming unit of virus per cell. CAT enzymatic activity can be detected in these cells as early as 4 hr after infection, reaching near-maximal levels at 48 hr. In fetal cells, CAT expression was maintained without a loss in activity for at least 1 week. Using in vitro studies as a guide, we introduced the AdCMVCATgD virus directly into adult rat myocardium and compared the expression results obtained from virus injection with those obtained by direct injection of pAdCMVCATgD plasmid DNA. The amount of CAT activity resulting from adenovirus infection of the myocardium was orders of magnitude higher than that seen from DNA injection and was proportional to the amount of input virus. Immunostaining for CAT protein in cardiac tissue sections following adenovirus injection demonstrated large numbers of positive cells, reaching nearly 100{\%} of the myocytes in many regions of the heart. Expression of genes introduced by adenovirus peaked at 5 days but was still detectable 55 days following infection. Adenoviruses are therefore a very useful tool for high-efficiency gene transfer into the cardiovascular system.",
author = "Alyson Kass-Eisler and Erik Falck-Pedersen and Mauricio Alvira and Johanna Rivera and Buttrick, {Peter M.} and Wittenberg, {Beatrice A.} and Laura Cipriani and Leinwand, {Leslie A.}",
year = "1993",
month = "12",
day = "15",
language = "English (US)",
volume = "90",
pages = "11498--11502",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "24",

}

TY - JOUR

T1 - Quantitative determination of adenovirus-mediated gene delivery to rat cardiac myocytes in vitro and in vivo

AU - Kass-Eisler, Alyson

AU - Falck-Pedersen, Erik

AU - Alvira, Mauricio

AU - Rivera, Johanna

AU - Buttrick, Peter M.

AU - Wittenberg, Beatrice A.

AU - Cipriani, Laura

AU - Leinwand, Leslie A.

PY - 1993/12/15

Y1 - 1993/12/15

N2 - To optimize the use of modified adenoviruses as vectors for gene delivery to the myocardium, we have characterized infection of cultured fetal and adult rat cardiac myocytes in vitro and of adult cardiac myocytes in vivo by using a replication-defective adenovirus carrying the chloramphenicol acetyltransferase (CAT) reporter gene driven by the cytomegalovirus promoter (AdCMVCATgD). In vitro, virtually all fetal or adult cardiocytes express the CAT gene when infected with 1 plaque-forming unit of virus per cell. CAT enzymatic activity can be detected in these cells as early as 4 hr after infection, reaching near-maximal levels at 48 hr. In fetal cells, CAT expression was maintained without a loss in activity for at least 1 week. Using in vitro studies as a guide, we introduced the AdCMVCATgD virus directly into adult rat myocardium and compared the expression results obtained from virus injection with those obtained by direct injection of pAdCMVCATgD plasmid DNA. The amount of CAT activity resulting from adenovirus infection of the myocardium was orders of magnitude higher than that seen from DNA injection and was proportional to the amount of input virus. Immunostaining for CAT protein in cardiac tissue sections following adenovirus injection demonstrated large numbers of positive cells, reaching nearly 100% of the myocytes in many regions of the heart. Expression of genes introduced by adenovirus peaked at 5 days but was still detectable 55 days following infection. Adenoviruses are therefore a very useful tool for high-efficiency gene transfer into the cardiovascular system.

AB - To optimize the use of modified adenoviruses as vectors for gene delivery to the myocardium, we have characterized infection of cultured fetal and adult rat cardiac myocytes in vitro and of adult cardiac myocytes in vivo by using a replication-defective adenovirus carrying the chloramphenicol acetyltransferase (CAT) reporter gene driven by the cytomegalovirus promoter (AdCMVCATgD). In vitro, virtually all fetal or adult cardiocytes express the CAT gene when infected with 1 plaque-forming unit of virus per cell. CAT enzymatic activity can be detected in these cells as early as 4 hr after infection, reaching near-maximal levels at 48 hr. In fetal cells, CAT expression was maintained without a loss in activity for at least 1 week. Using in vitro studies as a guide, we introduced the AdCMVCATgD virus directly into adult rat myocardium and compared the expression results obtained from virus injection with those obtained by direct injection of pAdCMVCATgD plasmid DNA. The amount of CAT activity resulting from adenovirus infection of the myocardium was orders of magnitude higher than that seen from DNA injection and was proportional to the amount of input virus. Immunostaining for CAT protein in cardiac tissue sections following adenovirus injection demonstrated large numbers of positive cells, reaching nearly 100% of the myocytes in many regions of the heart. Expression of genes introduced by adenovirus peaked at 5 days but was still detectable 55 days following infection. Adenoviruses are therefore a very useful tool for high-efficiency gene transfer into the cardiovascular system.

UR - http://www.scopus.com/inward/record.url?scp=0027133238&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027133238&partnerID=8YFLogxK

M3 - Article

C2 - 8265580

AN - SCOPUS:0027133238

VL - 90

SP - 11498

EP - 11502

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 24

ER -