A ligand-pseudoreceptor system based on de novo designed peptides for the generation of adenoviral vectors with altered tropism

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DOIResolve DOI: http://doi.org/10.1002/jgm.1155
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TypeArticle
Journal titleThe Journal of Gene Medicine
Volume10
Issue4
Pages355367; # of pages: 13
Subjectbio; Cell Line; Cells; Epidermal Growth Factor; Ligands; methods; Peptides; pha; Transgenes; adenovirus; retargeting; packaging cells; pseudoreceptor; gene therapy; fiber
AbstractBackground: Delivery of transgenes into specific tissues by adenovirus vectors (AdVs) relies on ablations of their natural tropism and on introduction of a new tropism. If the interaction with its natural receptor is ablated, a new packaging cell line is required to produce the AdV. In the present study, we have used two de novo designed peptides (E-Coil and K-Coil) that interact with each other with high affinity to establish a new receptor-ligand system for the propagation of retargeted AdVs. Methods: We produced a cell line (293E) expressing on its surface a pseudoreceptor containing the E-Coil. An AdV (AdFK4m/GFP) lacking the interaction with the primary receptor for adenovirus (CAR) and containing the K-Coil inserted at the fiber C-terminus was constructed and tested using two strategies: (1) an RGD motif (Arg-Gly-Asp) was inserted into the HI-loop of the fiber; (2) AdFK4m/GFP was conjugated to a bispecific adaptor for the epidermal growth factor receptor (EGFR). Results: AdFK4m/GFP infected 293E cells more efficiently than cells lacking the pseudoreceptor. The transduction was due to the K-Coil/E-Coil specific interaction since it was competed by addition of soluble K-Coil, but not soluble fiber. We demonstrated that the modified AdV was retargeted toward αv integrin by inclusion of the RGD motif, or toward EGFR using the bispecific adaptor. Conclusions: We have established a new system to produce AdVs ablated of natural tropism. This system should permit the retargeting of AdVs by inserting new ligandswithin the fiber or through the interactionwith bispecific adaptors.
Publication date
LanguageEnglish
AffiliationNational Research Council Canada; NRC Biotechnology Research Institute
Peer reviewedNo
NRC number47814
NPARC number3539700
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Record identifierb730416e-458b-48d9-b300-ecf4ae06776a
Record created2009-03-01
Record modified2016-05-09
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