Adenoviral-mediated gene transferinto the canine brain in vivo

Marianela Candolfi, Kurt M. Kroeger, G. Elizabeth Pluhar, Josee Bergeron, Mariana Puntel, James F. Curtin, Elizabeth A. McNiel, Andrew B. Freese, John R. Ohlfest, Peter F Moore, Pedro R. Lowenstein, Maria G. Castro

Research output: Contribution to journalArticle

11 Citations (Scopus)

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 β-galactosidase (βGal), 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 βGal (Ad-βGal), TK (Ad-TK), or Flt3L (Ad-Flt3L). We determined transgene expression by immunocytochemistry, βGal 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 βGal 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.

Original languageEnglish (US)
Pages (from-to)167-177
Number of pages11
JournalNeurosurgery
Volume60
Issue number1
DOIs
StatePublished - Jan 2007

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Galactosidases
Glioblastoma
Canidae
Thymidine Kinase
Dogs
Brain
Genes
Transgenes
Animal Models
Parietal Lobe
Injections
Human Herpesvirus 1
Microglia
Brain Neoplasms
Glioma
Rodentia
Cell Death
Enzyme-Linked Immunosorbent Assay
Immunohistochemistry
Macrophages

Keywords

  • Adenovirus
  • Dog
  • Flt3L
  • Ganciclovir
  • Gene therapy
  • Glioblastoma
  • HSV1-TK

ASJC Scopus subject areas

  • Surgery
  • Clinical Neurology

Cite this

Candolfi, M., Kroeger, K. M., Pluhar, G. E., Bergeron, J., Puntel, M., Curtin, J. F., ... Castro, M. G. (2007). Adenoviral-mediated gene transferinto the canine brain in vivo. Neurosurgery, 60(1), 167-177. https://doi.org/10.1227/01.NEU.0000249210.89096.6C

Adenoviral-mediated gene transferinto the canine brain in vivo. / 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 F; Lowenstein, Pedro R.; Castro, Maria G.

In: Neurosurgery, Vol. 60, No. 1, 01.2007, p. 167-177.

Research output: Contribution to journalArticle

Candolfi, M, Kroeger, KM, Pluhar, GE, Bergeron, J, Puntel, M, Curtin, JF, McNiel, EA, Freese, AB, Ohlfest, JR, Moore, PF, Lowenstein, PR & Castro, MG 2007, 'Adenoviral-mediated gene transferinto the canine brain in vivo', Neurosurgery, vol. 60, no. 1, pp. 167-177. https://doi.org/10.1227/01.NEU.0000249210.89096.6C
Candolfi M, Kroeger KM, Pluhar GE, Bergeron J, Puntel M, Curtin JF et al. Adenoviral-mediated gene transferinto the canine brain in vivo. Neurosurgery. 2007 Jan;60(1):167-177. https://doi.org/10.1227/01.NEU.0000249210.89096.6C
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 F ; Lowenstein, Pedro R. ; Castro, Maria G. / Adenoviral-mediated gene transferinto the canine brain in vivo. In: Neurosurgery. 2007 ; Vol. 60, No. 1. pp. 167-177.
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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 β-galactosidase (βGal), 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 βGal (Ad-βGal), TK (Ad-TK), or Flt3L (Ad-Flt3L). We determined transgene expression by immunocytochemistry, βGal 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 βGal 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.",
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AU - Curtin, James F.

AU - McNiel, Elizabeth A.

AU - Freese, Andrew B.

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N2 - 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 β-galactosidase (βGal), 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 βGal (Ad-βGal), TK (Ad-TK), or Flt3L (Ad-Flt3L). We determined transgene expression by immunocytochemistry, βGal 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 βGal 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.

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