Vaccinia as a vector for tumor-directed gene therapy: Biodistribution of a thymidine kinase-deleted mutant

Markus Puhlmann, Charles K. Brown, Michael Gnant, James Huang, Steven K. Libutti, H. Richard Alexander, David L. Bartlett

Research output: Contribution to journalArticle

123 Citations (Scopus)

Abstract

Tumor-directed gene therapy, such as 'suicide gene' therapy, requires high levels of gene expression in a high percentage of tumor cells in vivo to be effective. Current vector strategies have been ineffective in achieving these goals. This report introduces the attenuated (thymidine kinase (TK)- negative) replication-competent vaccinia virus (VV) as a potential vector for tumor-directed gene therapy by studying the biodistribution of VV in animal tumor models. A TK-deleted recombinant VV (Western Reserve strain) expressing luciferase on a synthetic promoter was constructed. Luciferase activity was measured in vitro after transduction of a variety of human and murine tumor cell lines and in vivo after intraperitoneal (i.p.) delivery in C57BL/6 mice with 7-day i.p. tumors (106 MC-38 cells). Three other in vivo tumor models were examined for tumor-specific gene expression after intravenous delivery of VV (human melanoma in nude mice, adenocarcinoma liver metastasis in immunocompetent mice, and subcutaneous sarcoma in the rat). In addition, a replication-incompetent vaccinia (1 μg of psoralen and ultraviolet light, 365 nm, 4 minutes) was tested in vitro and in vivo and compared with active virus. Luciferase activity in i.p. tumors at 4 days after i.p. injection of VV was >7000-fold higher than lung, >3000-fold higher than liver, and >250- fold higher than ovary. In addition, intravenous injection of VV resulted in markedly higher tumor luciferase activity compared with any other organ in every model tested (up to 188,000-fold higher than liver and 77,000-fold higher than lung). Inactivation of the virus resulted in negligible gene expression in vivo. In summary, VV has a high transduction efficiency in tumor cells with high levels of gene expression. The results suggest a selective in vivo replication of TK-deleted VV in tumor cells. Replication competent, TK-deleted VV appears to be an ideal vector for testing the in vivo delivery of toxic genes to tumor cells.

Original languageEnglish (US)
Pages (from-to)66-73
Number of pages8
JournalCancer Gene Therapy
Volume7
Issue number1
StatePublished - 2000
Externally publishedYes

Fingerprint

Vaccinia
Thymidine Kinase
Genetic Therapy
Vaccinia virus
Neoplasms
Luciferases
Gene Expression
Liver
Virus Inactivation
Ficusin
Lung
Poisons
Ultraviolet Rays
Tumor Cell Line
Intraperitoneal Injections
Inbred C57BL Mouse
Nude Mice
Intravenous Injections
Sarcoma
Suicide

Keywords

  • Biodistribution
  • Gene therapy
  • Luciferase
  • Thymidine kinase
  • Vaccinia

ASJC Scopus subject areas

  • Cancer Research
  • Genetics

Cite this

Puhlmann, M., Brown, C. K., Gnant, M., Huang, J., Libutti, S. K., Alexander, H. R., & Bartlett, D. L. (2000). Vaccinia as a vector for tumor-directed gene therapy: Biodistribution of a thymidine kinase-deleted mutant. Cancer Gene Therapy, 7(1), 66-73.

Vaccinia as a vector for tumor-directed gene therapy : Biodistribution of a thymidine kinase-deleted mutant. / Puhlmann, Markus; Brown, Charles K.; Gnant, Michael; Huang, James; Libutti, Steven K.; Alexander, H. Richard; Bartlett, David L.

In: Cancer Gene Therapy, Vol. 7, No. 1, 2000, p. 66-73.

Research output: Contribution to journalArticle

Puhlmann, M, Brown, CK, Gnant, M, Huang, J, Libutti, SK, Alexander, HR & Bartlett, DL 2000, 'Vaccinia as a vector for tumor-directed gene therapy: Biodistribution of a thymidine kinase-deleted mutant', Cancer Gene Therapy, vol. 7, no. 1, pp. 66-73.
Puhlmann M, Brown CK, Gnant M, Huang J, Libutti SK, Alexander HR et al. Vaccinia as a vector for tumor-directed gene therapy: Biodistribution of a thymidine kinase-deleted mutant. Cancer Gene Therapy. 2000;7(1):66-73.
Puhlmann, Markus ; Brown, Charles K. ; Gnant, Michael ; Huang, James ; Libutti, Steven K. ; Alexander, H. Richard ; Bartlett, David L. / Vaccinia as a vector for tumor-directed gene therapy : Biodistribution of a thymidine kinase-deleted mutant. In: Cancer Gene Therapy. 2000 ; Vol. 7, No. 1. pp. 66-73.
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