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Selected modification of the adenovirus type 5 hexon modulates interaction with coagulation factor X and hepatocyte transduction in vivo

Alba, R., Bradshaw, Angela C., Parker, Alan L., Bhella, D., Nicklin, Stuart A., Waddington, Simon N., Custers, Jerome, Goudsmit, J., Van Rooijen, Nico, Barouch, D. H., MacVey, John H. and Baker, Andrew H. 2009. Selected modification of the adenovirus type 5 hexon modulates interaction with coagulation factor X and hepatocyte transduction in vivo. Molecular Therapy 17 , S6-S6.

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Abstract

Recent studies have demonstrated the fundamental role of coagulation factors in adenovirus infectivity in vivo. It has been shown that coagulation factor X (FX) binds to the hypervariable regions (HVR) of the adenovirus serotype 5 (Ad5) hexon with high affinity. However, a systematic analysis of binding of FX to the HVRs to identify specific domains and amino acids critical for this interaction has not been performed. Here, cryo-electron microscopy of the Ad5:FX complex at 23Å was used to model the Ad5 hexon:FX interaction and identify contact residues in the hexon HVRs. In order to characterize the relative importance of these regions, we have generated novel hexon modified adenoviral vectors by homologous recombination via exchanging the hexon HVRs for those of adenoviral serotypes which do not bind FX, or by site directed mutagenesis of individual amino acids pinpointed by cryoelectron microscopy. All adenoviruses contained a CMV-lacZ reporter cassette and were amplified in HEK293 cells at similar titers to control Ad. Since the fiber and penton in each virus are identical, we assessed coxsackievirus and adenovirus receptor (CAR) dependent infection in HeLa or HepG2 cells showing all viruses transduced CAR permissive cells. Swapping either HVR5, HVR7 or both regions from adenovirus 5 to adenovirus 26 (which does not bind FX) reduced FX binding as analyzed by surface plasmon resonance, gene delivery in vitro and liver transduction in vivo. In addition, mutagenesis of amino acids in HVR5, HVR7 or both regions identified critical residues in each locale. We therefore show a critical role for HVR5 and HVR7 domains in mediating the Ad5 hexon:FX interaction. Novel Ad vectors with ablated FX interaction will be useful for future gene therapy applications where avoidance of FX-mediated effects is a key requirement.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Medicine
Subjects: Q Science > QR Microbiology
R Medicine > RM Therapeutics. Pharmacology
Publisher: Nature Publishing Group
Related URLs:
Last Modified: 04 Jun 2017 04:42
URI: http://orca.cf.ac.uk/id/eprint/43271

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