title: Differential susceptibility of primary and established human glioma cells to adenovirus infection: targeting via the epidermal growth factor receptor achieves fiber receptor-independent gene transfer.
abstract: Adenovirus (Ad) vectors are promising for gene therapy of glioma due to their ability to achieve efficient gene transfer upon intratumoral administration. Yet in this context, Ad mediates widespread gene transfer to both tumor and surrounding parenchyma. Ad entry is dependent upon the expression of fiber receptors, such as coxsackie/adenovirus receptor, and alpha(v) integrins on the target cells for binding and internalization, respectively. We hypothesized that the susceptibility of human gliomas to Ad would likely be heterogeneous due to variable expression of these receptors. It was found that established human glioma cell lines exhibited differential susceptibility to Ad-mediated gene transfer, which correlated directly with the level of radiolabeled Ad binding and with the expression of coxsackie/adenovirus receptor but not with the expression of alpha(v) integrins. To circumvent the lack of fiber receptors and to target Ad gene transfer specifically to tumor cells, we used a bispecific antibody conjugate to ablate Ad binding to fiber receptors and retarget binding to the epidermal growth factor receptor (EGFR), a tumor-associated marker negligibly expressed in normal, mitotically quiescent neural tissues. The results demonstrate that EGFR-targeted Ad gene transfer was EGFR specific and independent of fiber-fiber receptor interactions. Furthermore, EGFR targeting significantly enhanced Ad gene delivery to 7 of 12 established glioma cell lines and to 6 of 8 cultured primary gliomas. Interestingly, EGFR-targeted Ad gene transfer did not correlate with EGFR expression across cell lines, suggesting the importance of other factors. This study establishes that fiber receptor expression limits the utility of Ad vectors for gene transfer to glioma cells and suggests that targeting Ad via EGFR may prove valuable for tumor-specific gene transfer to high-grade gliomas. These findings have key relevance in the context of Ad vector-based approaches for glioma gene therapy.
authors.affiliation: Gene Therapy Program, University of Alabama at Birmingham, 35294, USA.
authors.forename: C R
authors.initials: CR
authors.lastname: Miller
authors.forename: D J
authors.initials: DJ
authors.lastname: Buchsbaum
authors.forename: P N
authors.initials: PN
authors.lastname: Reynolds
authors.forename: J T
authors.initials: JT
authors.lastname: Douglas
authors.forename: G Y
authors.initials: GY
authors.lastname: Gillespie
authors.forename: M S
authors.initials: MS
authors.lastname: Mayo
authors.forename: D
authors.initials: D
authors.lastname: Raben
authors.forename: D T
authors.initials: DT
authors.lastname: Curiel
mesh_terms: D000260:Adenoviruses, Human / Q000235:genetics; D018033:Antibodies, Bispecific / Q000008:administration & dosage; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D066246:ErbB Receptors / Q000276:immunology* / Q000378:metabolism; D018390:Gene Targeting; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D005910:Glioma / Q000235:genetics*; D006801:Humans; D018796:Immunoconjugates; D011991:Receptors, Virus / Q000276:immunology / Q000378:metabolism; D014407:Tumor Cells, Cultured
chemical_list: D018033:Antibodies, Bispecific; C568771:CLMP protein, human; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D018796:Immunoconjugates; D011991:Receptors, Virus; D066246:ErbB Receptors