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

TitleQuantitative determination of adenovirus-mediated gene delivery to rat cardiac myocytes in vitro and in vivo.
Publication TypeJournal Article
Year of Publication1993
AuthorsKass-Eisler A, Falck-Pedersen E, Alvira M, Rivera J, Buttrick PM, Wittenberg BA, Cipriani L, Leinwand LA
JournalProc Natl Acad Sci U S A
Volume90
Issue24
Pagination11498-502
Date Published1993 Dec 15
ISSN0027-8424
KeywordsAnimals, Cell Line, Cells, Cultured, Chloramphenicol O-Acetyltransferase, Cytomegalovirus, DNA, Bacterial, Female, Fetus, Gene Transfer Techniques, Genetic Vectors, Heart, Humans, Immunohistochemistry, Kidney, Myocardium, Promoter Regions, Genetic, Rats, Rats, Sprague-Dawley
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.

DOI10.1073/pnas.90.24.11498
Alternate JournalProc Natl Acad Sci U S A
PubMed ID8265580
PubMed Central IDPMC48011
Grant ListGM29090 / GM / NIGMS NIH HHS / United States
GM41967 / GM / NIGMS NIH HHS / United States
T32AG0092-05 / AG / NIA NIH HHS / United States

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