In vivo expression of prostate-specific adenoviral vectors in a canine model

Mitchell S. Steiner, Yu Zhang, John Carraher, Yi Lu

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

The activity and expression of transgene β-galactosidase (lacZ) by replication-deficient adenoviral vectors (Ad-lacZ) containing prostate-specific promoters were compared using an in vivo canine model. The prostate tissue-specific promoters were prostate-specific antigen, probasin, and mouse mammary tumor virus long-terminal repeat, which were fused separately to an Escherichia coli lacZ gene. Dogs underwent laparotomy, and adenoviral vectors were delivered by direct intraprostatic injection. At 72 hours postinjection, the prostate and various other organs were harvested to evaluate the degree of prostate expression and dissemination of adenoviral vectors. Expression of lacZ in tissues was determined by 5-bromo-4-chloro-3-indolyl β-D-galactoside staining, β-galactosidase assay, and E. coli lacZ reverse transcriptase-polymerase chain reaction (PCR). The presence of adenoviral DNA sequences in canine tissues was determined by PCR using primers specific for the type 5 adenoviral genome. All three of the prostate-specific adenoviruses tested effectively expressed the lacZ gene in the canine prostate, but expression levels were lower than that of the control viral vector AdRSVlacZ following intraprostatic injection. By PCR, adenoviral vector DNA was detected in other organs and tissues, including the bladder and vas deferens. However, reverse transcriptase-PCR analysis revealed that prostate-specific Ad-lacZ vectors only transcribed lacZ mRNA in the prostate and not in nonprostatic tissues. Thus, these novel prostate-specific adenoviral vectors each have equal in vivo expression exclusively in the prostate and may potentially be used for prostate cancer gene therapy.

Original languageEnglish (US)
Pages (from-to)456-464
Number of pages9
JournalCancer Gene Therapy
Volume6
Issue number5
DOIs
StatePublished - Jan 1 1999

Fingerprint

Canidae
Prostate
Galactosidases
Lac Operon
Reverse Transcriptase Polymerase Chain Reaction
Escherichia coli
Mouse mammary tumor virus
Galactosides
Polymerase Chain Reaction
Injections
Vas Deferens
Terminal Repeat Sequences
Neoplasm Genes
Prostate-Specific Antigen
Transgenes
Adenoviridae
Genetic Therapy
Laparotomy
Prostatic Neoplasms
Urinary Bladder

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Molecular Biology
  • Cancer Research

Cite this

In vivo expression of prostate-specific adenoviral vectors in a canine model. / Steiner, Mitchell S.; Zhang, Yu; Carraher, John; Lu, Yi.

In: Cancer Gene Therapy, Vol. 6, No. 5, 01.01.1999, p. 456-464.

Research output: Contribution to journalArticle

Steiner, Mitchell S. ; Zhang, Yu ; Carraher, John ; Lu, Yi. / In vivo expression of prostate-specific adenoviral vectors in a canine model. In: Cancer Gene Therapy. 1999 ; Vol. 6, No. 5. pp. 456-464.
@article{991fc3c6fb334c1c95d035fc3ab2f977,
title = "In vivo expression of prostate-specific adenoviral vectors in a canine model",
abstract = "The activity and expression of transgene β-galactosidase (lacZ) by replication-deficient adenoviral vectors (Ad-lacZ) containing prostate-specific promoters were compared using an in vivo canine model. The prostate tissue-specific promoters were prostate-specific antigen, probasin, and mouse mammary tumor virus long-terminal repeat, which were fused separately to an Escherichia coli lacZ gene. Dogs underwent laparotomy, and adenoviral vectors were delivered by direct intraprostatic injection. At 72 hours postinjection, the prostate and various other organs were harvested to evaluate the degree of prostate expression and dissemination of adenoviral vectors. Expression of lacZ in tissues was determined by 5-bromo-4-chloro-3-indolyl β-D-galactoside staining, β-galactosidase assay, and E. coli lacZ reverse transcriptase-polymerase chain reaction (PCR). The presence of adenoviral DNA sequences in canine tissues was determined by PCR using primers specific for the type 5 adenoviral genome. All three of the prostate-specific adenoviruses tested effectively expressed the lacZ gene in the canine prostate, but expression levels were lower than that of the control viral vector AdRSVlacZ following intraprostatic injection. By PCR, adenoviral vector DNA was detected in other organs and tissues, including the bladder and vas deferens. However, reverse transcriptase-PCR analysis revealed that prostate-specific Ad-lacZ vectors only transcribed lacZ mRNA in the prostate and not in nonprostatic tissues. Thus, these novel prostate-specific adenoviral vectors each have equal in vivo expression exclusively in the prostate and may potentially be used for prostate cancer gene therapy.",
author = "Steiner, {Mitchell S.} and Yu Zhang and John Carraher and Yi Lu",
year = "1999",
month = "1",
day = "1",
doi = "10.1038/sj.cgt.7700065",
language = "English (US)",
volume = "6",
pages = "456--464",
journal = "Cancer Gene Therapy",
issn = "0929-1903",
publisher = "Nature Publishing Group",
number = "5",

}

TY - JOUR

T1 - In vivo expression of prostate-specific adenoviral vectors in a canine model

AU - Steiner, Mitchell S.

AU - Zhang, Yu

AU - Carraher, John

AU - Lu, Yi

PY - 1999/1/1

Y1 - 1999/1/1

N2 - The activity and expression of transgene β-galactosidase (lacZ) by replication-deficient adenoviral vectors (Ad-lacZ) containing prostate-specific promoters were compared using an in vivo canine model. The prostate tissue-specific promoters were prostate-specific antigen, probasin, and mouse mammary tumor virus long-terminal repeat, which were fused separately to an Escherichia coli lacZ gene. Dogs underwent laparotomy, and adenoviral vectors were delivered by direct intraprostatic injection. At 72 hours postinjection, the prostate and various other organs were harvested to evaluate the degree of prostate expression and dissemination of adenoviral vectors. Expression of lacZ in tissues was determined by 5-bromo-4-chloro-3-indolyl β-D-galactoside staining, β-galactosidase assay, and E. coli lacZ reverse transcriptase-polymerase chain reaction (PCR). The presence of adenoviral DNA sequences in canine tissues was determined by PCR using primers specific for the type 5 adenoviral genome. All three of the prostate-specific adenoviruses tested effectively expressed the lacZ gene in the canine prostate, but expression levels were lower than that of the control viral vector AdRSVlacZ following intraprostatic injection. By PCR, adenoviral vector DNA was detected in other organs and tissues, including the bladder and vas deferens. However, reverse transcriptase-PCR analysis revealed that prostate-specific Ad-lacZ vectors only transcribed lacZ mRNA in the prostate and not in nonprostatic tissues. Thus, these novel prostate-specific adenoviral vectors each have equal in vivo expression exclusively in the prostate and may potentially be used for prostate cancer gene therapy.

AB - The activity and expression of transgene β-galactosidase (lacZ) by replication-deficient adenoviral vectors (Ad-lacZ) containing prostate-specific promoters were compared using an in vivo canine model. The prostate tissue-specific promoters were prostate-specific antigen, probasin, and mouse mammary tumor virus long-terminal repeat, which were fused separately to an Escherichia coli lacZ gene. Dogs underwent laparotomy, and adenoviral vectors were delivered by direct intraprostatic injection. At 72 hours postinjection, the prostate and various other organs were harvested to evaluate the degree of prostate expression and dissemination of adenoviral vectors. Expression of lacZ in tissues was determined by 5-bromo-4-chloro-3-indolyl β-D-galactoside staining, β-galactosidase assay, and E. coli lacZ reverse transcriptase-polymerase chain reaction (PCR). The presence of adenoviral DNA sequences in canine tissues was determined by PCR using primers specific for the type 5 adenoviral genome. All three of the prostate-specific adenoviruses tested effectively expressed the lacZ gene in the canine prostate, but expression levels were lower than that of the control viral vector AdRSVlacZ following intraprostatic injection. By PCR, adenoviral vector DNA was detected in other organs and tissues, including the bladder and vas deferens. However, reverse transcriptase-PCR analysis revealed that prostate-specific Ad-lacZ vectors only transcribed lacZ mRNA in the prostate and not in nonprostatic tissues. Thus, these novel prostate-specific adenoviral vectors each have equal in vivo expression exclusively in the prostate and may potentially be used for prostate cancer gene therapy.

UR - http://www.scopus.com/inward/record.url?scp=0033194877&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033194877&partnerID=8YFLogxK

U2 - 10.1038/sj.cgt.7700065

DO - 10.1038/sj.cgt.7700065

M3 - Article

VL - 6

SP - 456

EP - 464

JO - Cancer Gene Therapy

JF - Cancer Gene Therapy

SN - 0929-1903

IS - 5

ER -