Comparison of viral vectors: Gene transfer efficiency and tissue specificity in a bladder cancer model

D. Robert Siemens, Scott Crist, James (Christopher) Austin, James Tartaglia, Timothy L. Ratliff

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Purpose: Gene transfer efficiency and specific cell targeting of vectors is a major obstacle in preclinical studies of gene therapy for malignant disease. Previous attempts at gene transfer in bladder cancer models have resulted in variable urothelial and tumor transgene expression after intravesical administration of recombinant viral vectors. In the current study we compared the gene transfer efficiencies of different viral vectors. Materials and Methods: We compared the gene transfer efficiencies of the viral vectors replication-deficient adenovirus, attenuated vaccinia virus (NYVAC) and canarypox virus (ALVAC) in vitro and in an orthotopic murine bladder cancer model. We used β-galactosidase and firefly luciferase reporter gene expression to compare gene transfer efficiency. Results: Significantly higher transgene expression was observed in vitro when these cells were infected with NYVAC or ALVAC compared with adenovirus vectors. Similarly the efficiency of adenovirus vectors to transfer genetic material into bladder urothelium and orthotopic bladder tumors was inferior to that of ALVAC and NYVAC vectors, which interestingly appeared to have a predilection to infect the orthotopic tumor. Analysis of the expression of coxsackie-adenovirus receptor using reverse transcriptase-polymerase chain reaction revealed the bladder tumor cell lines were lacking this adenovirus receptor. While adenovirus transferred genes poorly to normal bladder, coxsackie-adenovirus receptor expression was high in bladder tissue. Conclusions: The viral vectors examined in these experiments resulted in significantly different gene transfer in the orthotopic bladder cancer model, underscoring the importance of vector selection in gene therapy protocols.

Original languageEnglish (US)
Pages (from-to)979-984
Number of pages6
JournalJournal of Urology
Volume170
Issue number3
DOIs
StatePublished - Sep 1 2003
Externally publishedYes

Fingerprint

Organ Specificity
Viral Genes
Urinary Bladder Neoplasms
Adenoviridae
Genes
Coxsackie and Adenovirus Receptor-Like Membrane Protein
Urinary Bladder
Transgenes
Genetic Therapy
Canarypox virus
Galactosidases
Intravesical Administration
Firefly Luciferases
Urothelium
Vaccinia virus
Tumor Cell Line
Reverse Transcriptase Polymerase Chain Reaction
Reporter Genes
Neoplasms
Gene Expression

Keywords

  • Bladder
  • Bladder neoplasms
  • Gene therapy
  • Gene transfer techniques
  • Genetic vectors

ASJC Scopus subject areas

  • Urology

Cite this

Comparison of viral vectors : Gene transfer efficiency and tissue specificity in a bladder cancer model. / Siemens, D. Robert; Crist, Scott; Austin, James (Christopher); Tartaglia, James; Ratliff, Timothy L.

In: Journal of Urology, Vol. 170, No. 3, 01.09.2003, p. 979-984.

Research output: Contribution to journalArticle

Siemens, D. Robert ; Crist, Scott ; Austin, James (Christopher) ; Tartaglia, James ; Ratliff, Timothy L. / Comparison of viral vectors : Gene transfer efficiency and tissue specificity in a bladder cancer model. In: Journal of Urology. 2003 ; Vol. 170, No. 3. pp. 979-984.
@article{197fffe7a89946049cc8169a631a235b,
title = "Comparison of viral vectors: Gene transfer efficiency and tissue specificity in a bladder cancer model",
abstract = "Purpose: Gene transfer efficiency and specific cell targeting of vectors is a major obstacle in preclinical studies of gene therapy for malignant disease. Previous attempts at gene transfer in bladder cancer models have resulted in variable urothelial and tumor transgene expression after intravesical administration of recombinant viral vectors. In the current study we compared the gene transfer efficiencies of different viral vectors. Materials and Methods: We compared the gene transfer efficiencies of the viral vectors replication-deficient adenovirus, attenuated vaccinia virus (NYVAC) and canarypox virus (ALVAC) in vitro and in an orthotopic murine bladder cancer model. We used β-galactosidase and firefly luciferase reporter gene expression to compare gene transfer efficiency. Results: Significantly higher transgene expression was observed in vitro when these cells were infected with NYVAC or ALVAC compared with adenovirus vectors. Similarly the efficiency of adenovirus vectors to transfer genetic material into bladder urothelium and orthotopic bladder tumors was inferior to that of ALVAC and NYVAC vectors, which interestingly appeared to have a predilection to infect the orthotopic tumor. Analysis of the expression of coxsackie-adenovirus receptor using reverse transcriptase-polymerase chain reaction revealed the bladder tumor cell lines were lacking this adenovirus receptor. While adenovirus transferred genes poorly to normal bladder, coxsackie-adenovirus receptor expression was high in bladder tissue. Conclusions: The viral vectors examined in these experiments resulted in significantly different gene transfer in the orthotopic bladder cancer model, underscoring the importance of vector selection in gene therapy protocols.",
keywords = "Bladder, Bladder neoplasms, Gene therapy, Gene transfer techniques, Genetic vectors",
author = "Siemens, {D. Robert} and Scott Crist and Austin, {James (Christopher)} and James Tartaglia and Ratliff, {Timothy L.}",
year = "2003",
month = "9",
day = "1",
doi = "10.1097/01.ju.0000070925.10039.23",
language = "English (US)",
volume = "170",
pages = "979--984",
journal = "Journal of Urology",
issn = "0022-5347",
publisher = "Elsevier Inc.",
number = "3",

}

TY - JOUR

T1 - Comparison of viral vectors

T2 - Gene transfer efficiency and tissue specificity in a bladder cancer model

AU - Siemens, D. Robert

AU - Crist, Scott

AU - Austin, James (Christopher)

AU - Tartaglia, James

AU - Ratliff, Timothy L.

PY - 2003/9/1

Y1 - 2003/9/1

N2 - Purpose: Gene transfer efficiency and specific cell targeting of vectors is a major obstacle in preclinical studies of gene therapy for malignant disease. Previous attempts at gene transfer in bladder cancer models have resulted in variable urothelial and tumor transgene expression after intravesical administration of recombinant viral vectors. In the current study we compared the gene transfer efficiencies of different viral vectors. Materials and Methods: We compared the gene transfer efficiencies of the viral vectors replication-deficient adenovirus, attenuated vaccinia virus (NYVAC) and canarypox virus (ALVAC) in vitro and in an orthotopic murine bladder cancer model. We used β-galactosidase and firefly luciferase reporter gene expression to compare gene transfer efficiency. Results: Significantly higher transgene expression was observed in vitro when these cells were infected with NYVAC or ALVAC compared with adenovirus vectors. Similarly the efficiency of adenovirus vectors to transfer genetic material into bladder urothelium and orthotopic bladder tumors was inferior to that of ALVAC and NYVAC vectors, which interestingly appeared to have a predilection to infect the orthotopic tumor. Analysis of the expression of coxsackie-adenovirus receptor using reverse transcriptase-polymerase chain reaction revealed the bladder tumor cell lines were lacking this adenovirus receptor. While adenovirus transferred genes poorly to normal bladder, coxsackie-adenovirus receptor expression was high in bladder tissue. Conclusions: The viral vectors examined in these experiments resulted in significantly different gene transfer in the orthotopic bladder cancer model, underscoring the importance of vector selection in gene therapy protocols.

AB - Purpose: Gene transfer efficiency and specific cell targeting of vectors is a major obstacle in preclinical studies of gene therapy for malignant disease. Previous attempts at gene transfer in bladder cancer models have resulted in variable urothelial and tumor transgene expression after intravesical administration of recombinant viral vectors. In the current study we compared the gene transfer efficiencies of different viral vectors. Materials and Methods: We compared the gene transfer efficiencies of the viral vectors replication-deficient adenovirus, attenuated vaccinia virus (NYVAC) and canarypox virus (ALVAC) in vitro and in an orthotopic murine bladder cancer model. We used β-galactosidase and firefly luciferase reporter gene expression to compare gene transfer efficiency. Results: Significantly higher transgene expression was observed in vitro when these cells were infected with NYVAC or ALVAC compared with adenovirus vectors. Similarly the efficiency of adenovirus vectors to transfer genetic material into bladder urothelium and orthotopic bladder tumors was inferior to that of ALVAC and NYVAC vectors, which interestingly appeared to have a predilection to infect the orthotopic tumor. Analysis of the expression of coxsackie-adenovirus receptor using reverse transcriptase-polymerase chain reaction revealed the bladder tumor cell lines were lacking this adenovirus receptor. While adenovirus transferred genes poorly to normal bladder, coxsackie-adenovirus receptor expression was high in bladder tissue. Conclusions: The viral vectors examined in these experiments resulted in significantly different gene transfer in the orthotopic bladder cancer model, underscoring the importance of vector selection in gene therapy protocols.

KW - Bladder

KW - Bladder neoplasms

KW - Gene therapy

KW - Gene transfer techniques

KW - Genetic vectors

UR - http://www.scopus.com/inward/record.url?scp=0043239038&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0043239038&partnerID=8YFLogxK

U2 - 10.1097/01.ju.0000070925.10039.23

DO - 10.1097/01.ju.0000070925.10039.23

M3 - Article

C2 - 12913754

AN - SCOPUS:0043239038

VL - 170

SP - 979

EP - 984

JO - Journal of Urology

JF - Journal of Urology

SN - 0022-5347

IS - 3

ER -