Adenoviral Mediated Gene Transfer into the Dog Brain In Vivo

Authors

Marianela Candolfi, Cedars-Sinai Medical Center
Kurt Kroeger, Cedars-Sinai Medical Center
Elizabeth Pluhar, University of Minnesota - Twin Cities
Chunyan Liu, Cedars-Sinai Medical Center
Carlos Barcia, Cedars-Sinai Medical Center
Josee Bergeron, Cedars-Sinai Medical Center
Mariana Puntel, Cedars-Sinai Medical Center
James Curtin, Technological University DublinFollow
Elizabeth McNiel, University of Minnesota - Twin Cities
Andrew Freese, University of Minnesota - Twin Cities
John Ohlfest, University of Minnesota - Twin Cities
Peter Moore, University of California - Davis
William Kuoy, Cedars-Sinai Medical Center
Pedro Lowenstein, Cedars-Sinai Medical Center
Maria Castro, Cedars-Sinai Medical Center

Document Type

Article

Rights

Available under a Creative Commons Attribution Non-Commercial Share Alike 4.0 International Licence

Disciplines

1.6 BIOLOGICAL SCIENCES

Publication Details

Neurosurgery. 60(1):167-178, January 2007. doi: 10.1227/01.NEU.0000249210.89096.6C

Abstract

OBJECTIVE: Glioblastoma multiforme (GBM) is a devastating brain tumor for which there is no cure. Adenoviral-mediated transfer of conditional cytotoxic (herpes simplex virus [HSV] 1-derived thymidine kinase [TK]) and immunostimulatory (Fms-like tyrosine kinase 3 ligand [Flt3L]) transgenes elicited immune-mediated long-term survival in a syngeneic intracranial GBM model in rodents. However, the lack of a large GBM animal model makes it difficult to predict the outcome of therapies in humans. Dogs develop spontaneous GBM that closely resemble the human disease; therefore, they constitute an excellent large animal model. We assayed the transduction efficiency of adenoviral vectors (Ads) encoding beta-galactosidase (betaGal), TK, and Flt3L in J3T dog GBM cells in vitro and in the dog brain in vivo. METHODS: J3T cells were infected with Ads (30 plaque-forming units/cell; 72 h) encoding betaGal (Ad-betaGal), TK (Ad-TK), or Flt3L (Ad-Flt3L). We determined transgene expression by immunocytochemistry, betaGal activity, Flt3L enzyme-linked immunosorbent assay, and TK-induced cell death. Ads were also injected intracranially into the parietal cortex of healthy dogs. We determined cell-type specific transgene expression and immune cell infiltration. RESULTS: Adenoviral-mediated gene transfer of HSV1-TK, Flt3L, and betaGal was detected in dog glioma cells in vitro (45% transduction efficiency) and in the dog brain in vivo (10-mm area transduced surrounding each injection site). T cells and macrophages/activated microglia infiltrated the injection sites. Importantly, no adverse clinical or neuropathological side effects were observed. CONCLUSION: We demonstrate effective adenoviral-mediated gene transfer into the brain of dogs in vivo and support the use of these vectors to develop an efficacy trial for canine GBM as a prelude to human trials.

DOI

10.1227/01.NEU.0000249210.89096.6C

Recommended Citation

Candolfi, Marianela; Kroeger, Kurt M.; Pluhar, G. Elizabeth; Bergeron, Josee; Puntel, Mariana; Curtin, James F.; McNiel, Elizabeth A.; Freese, Andrew B.; Ohlfest, John R.; Moore, Peter; Lowenstein, Pedro R.; Castro, Maria G. (2007) Adenoviral-Mediated Gene Transferinto the Canine Brain in Vivo. Neurosurgery. 60(1):167-178, January 2007. doi:10.1227/01.NEU.0000249210.89096.6C

Funder

National Institute of Health, Bram and Elaine Goldsmith Chair in Gene Therapeutics, The Linda Tallen and David Paul Kane Annual Fellowship, Board of Governors at Cedars Sinai Medical Center.