Novel recombinant adenoviral vector that targets the interleukin-13 receptor alpha2 chain permits effective gene transfer to malignant glioma.
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.
Ulasov
Ilya V
IV
Division of Neurosurgery, University of Chicago, Chicago, IL 60637, USA.
Tyler
Matthew A
MA
Han
Yu
Y
Glasgow
Joel N
JN
Lesniak
Maciej S
MS
D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D004274:DNA, Recombinant / Q000235:genetics / Q000378:metabolism*; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D005910:Glioma / Q000235:genetics* / Q000378:metabolism / Q000628:therapy*; D006801:Humans; D018793:Interleukin-13 / Q000378:metabolism; D053663:Interleukin-13 Receptor alpha2 Subunit / Q000378:metabolism*; D008297:Male; D051379:Mice; D008819:Mice, Nude; D018613:Microscopy, Confocal; D015203:Reproducibility of Results; D014161:Transduction, Genetic; D019076:Transgenes; D014764:Viral Proteins / Q000378:metabolism; D014779:Virus Replication; D023041:Xenograft Model Antitumor Assays
D004274:DNA, Recombinant; D018793:Interleukin-13; D053663:Interleukin-13 Receptor alpha2 Subunit; D014764:Viral Proteins; C057839:fibritin protein, Enterobacteria phage T4