Novel recombinant adenoviral vector that targets the interleukin-13 receptor alpha2 chain permits effective gene transfer to malignant glioma.

Abstract

Transduction of malignant glioma with adenovirus serotype 5 (Ad5) vectors is limited by the low levels of coxsackievirus and adenovirus receptor (CAR) on tumor cells. However, malignant brain tumors have been found to overexpress a glioma-associated receptor, interleukin-13 receptor alpha2 chain (IL-13Ralpha2), a marker of both glial transformation and tumor grade. To selectively target Ad5 to IL-13Ralpha2, we constructed a replication-deficient adenoviral vector that possesses an IL-13 ligand presented by a T4 phage fibritin shaft, and designated the new virus LU-13. Western blot and sequence analyses confirmed proper trimerization and ligand presentation by the T4 fibritin shaft. Confocal microscopy analysis of primary glioma suspensions incubated with viral recombinants showed that LU-13 colocalized with IL-13Ralpha2. Luciferase transduction assays conducted in both primary and passaged glioma cell cultures exhibited at least 10-fold enhanced gene transduction. Moreover, the virus preferentially bound to glioma cells, as documented by increased adenoviral E4 DNA copy number. In vitro competition assays performed with anti-human IL-13 monoclonal antibody confirmed significant attenuation of LU-13 transduction. These results were further confirmed in vivo, where LU-13 showed a 300-fold increase in transgene expression. In summary, we describe here the development of a novel and targeted adenoviral vector that binds IL-13Ralpha2. Our findings confirm the ability of LU-13 to bind IL-13Ralpha2 and increase transgene expression, making it an attractive gene therapy vector for the treatment of malignant glioma in a clinical setting.

• Authors: Ilya V Ulasov, Matthew A Tyler, Yu Han, Joel N Glasgow, Maciej S Lesniak
• Journal: Human gene therapy (Hum Gene Ther) Vol. 18 Issue 2 Pg. 118-29 (Feb 2007) ISSN: 1043-0342 [Print] United States
• PMID: 17328684 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• DNA, Recombinant
• Interleukin-13
• Interleukin-13 Receptor alpha2 Subunit
• Viral Proteins
• fibritin protein, Enterobacteria phage T4

Topics

• Adenoviridae (genetics, metabolism)
• Animals
• DNA, Recombinant (genetics, metabolism)
• Genetic Engineering
• Genetic Therapy (methods)
• Genetic Vectors (genetics)
• Glioma (genetics, metabolism, therapy)
• Humans
• Interleukin-13 (metabolism)
• Interleukin-13 Receptor alpha2 Subunit (metabolism)
• Male
• Mice
• Mice, Nude
• Microscopy, Confocal
• Reproducibility of Results
• Transduction, Genetic
• Transgenes
• Viral Proteins (metabolism)
• Virus Replication
• Xenograft Model Antitumor Assays