Use of quantitative TaqMan real-time PCR to track the time-dependent distribution of gene transfer vectors in vivo

Neil R. Hackett, Tarek B. Sawy, Leonard Y. Lee, Ivan Silva, John O'Leary, Todd K. Rosengart, Ronald Crystal

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

To assess the biodistribution and pharmacokinetics of gene transfer vectors, real-time PCR with fluorescent TaqMan chemistry was used to quantify tissue levels of adenovirus gene transfer vectors (Ad) following myocardial administration. After optimizing the detection of the genome of Ad vectors expressing human vascular endothelial growth factor (AdGVVEGF121.10) and Escherichia coli cytosine deaminase (AdGVCD.10), a comparison was made of intramyocardial injection versus intracoronary delivery to the left ventricle of the pig. One hour post-intramyocardial administration, the left ventricular Ad genome level was 6.2 copies per cellular genome, 26-fold higher than the level of 0.24 copies per cellular genome following intracoronary administration. Relative to the vector levels after 1 h, the amount dropped 14-and 5.5-fold by 24 h following intramyocardial and intracoronary administration, respectively. Interestingly, the vector that escaped the left ventricle after intracoronary or intramyocardial administration to pigs was found primarily within the lung, an observation in marked variance to the biodistribution of Ad vector in rodents. In this regard, after intravenous injection to the pig, 90% of the recovered vector was found in the lung, and even after intrahepatic portal vein injection, 55% of the recovered vector was in the lung. These data have important implications regarding the use of experimental animals for safety studies on administration of Ad to humans.

Original languageEnglish
Pages (from-to)649-656
Number of pages8
JournalMolecular Therapy
Volume2
Issue number6
DOIs
Publication statusPublished - 1 Jan 2000
Externally publishedYes

Fingerprint

Real-Time Polymerase Chain Reaction
Genome
Swine
Lung
Genes
Heart Ventricles
Cytosine Deaminase
Injections
Portal Vein
Adenoviridae
Intravenous Injections
Rodentia
Pharmacokinetics
Observation
Escherichia coli
Safety

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Pharmacology
  • Drug Discovery

Cite this

Use of quantitative TaqMan real-time PCR to track the time-dependent distribution of gene transfer vectors in vivo. / Hackett, Neil R.; Sawy, Tarek B.; Lee, Leonard Y.; Silva, Ivan; O'Leary, John; Rosengart, Todd K.; Crystal, Ronald.

In: Molecular Therapy, Vol. 2, No. 6, 01.01.2000, p. 649-656.

Research output: Contribution to journalArticle

Hackett, Neil R. ; Sawy, Tarek B. ; Lee, Leonard Y. ; Silva, Ivan ; O'Leary, John ; Rosengart, Todd K. ; Crystal, Ronald. / Use of quantitative TaqMan real-time PCR to track the time-dependent distribution of gene transfer vectors in vivo. In: Molecular Therapy. 2000 ; Vol. 2, No. 6. pp. 649-656.
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