Influence of adenoviral fiber mutations on viral encapsidation, infectivity and in vivo tropism

Leissner, Philippe; Legrand, Valérie; Schlesinger, Yasmin; Hadji, D.A.; Raaij, Mark J. van; Cusack, Stephen; Pavirani, Andréa; Mehtali, Majid

doi:10.1038/sj.gt.3301343

Cited by 90 publications

(75 citation statements)

References 46 publications

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“…One way to do this rationally is by designing point mutants that disrupt undesired binding to certain receptors and promote the binding to others. To this end, elimination of CAR binding by HAd5 fiber has already been achieved (71,72). In this work, we identify point mutants of CAV-2 and HAd37 fiber heads with greatly reduced ability to bind immobilized CAR D1.…”

mentioning

confidence: 97%

Structural and Mutational Analysis of Human Ad37 and Canine Adenovirus 2 Fiber Heads in Complex with the D1 Domain of Coxsackie and Adenovirus Receptor

Seiradake

Lortat‐Jacob

Billet

et al. 2006

Journal of Biological Chemistry

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Adenovirus fibers from most serotypes bind the D1 domain of coxsackie and adenovirus receptor (CAR), although the binding residues are not strictly conserved. To understand this further, we determined the crystal structures of canine adenovirus serotype 2 (CAV-2) and the human adenovirus serotype 37 (HAd37) in complex with human CAR D1 at 2.3 and 1.5 Å resolution, respectively. Structure comparison with the HAd12 fiber head-CAR D1 complex showed that the overall topology of the interaction is conserved but that the interfaces differ in number and identity of interacting residues, shape complementarity, and degree of conformational adaptation. Using surface plasmon resonance, we characterized the binding affinity to CAR D1 of wild type and mutant CAV-2 and HAd37 fiber heads. We found that CAV-2 has the highest affinity but fewest direct interactions, with the reverse being true for HAd37. Moreover, we found that conserved interactions can have a minor contribution, whereas serotype-specific interactions can be essential. These results are discussed in the light of virus evolution and design of adenovirus vectors for gene transfer. Adenoviruses (Ads)5 are nonenveloped icosahedral particles (70 -90 nm in diameter) containing double-stranded DNA genomes of 28 -42 kbp. They have been isolated from a wide variety of vertebrates, including mammals, birds, reptiles, amphibians, and fish (1). The ϳ50 human Ad serotypes (HAd) (divided into species A-F) lead to serotype-specific respiratory, ocular, and enteric infections that are usually self-limiting diseases in immunocompetent individuals. However, Ad infections are a significant cause of morbidity and mortality in newborns and immunosuppressed individuals (2). Notably, bone marrow and solid organ transplant patients who have severe lymphocytopenia are at the highest risk for Ad-induced disease (3). In addition, there are no drugs currently available that efficiently prevent or treat infections from all HAd serotypes.The major adenovirus capsid protein is the trimeric hexon, 240 of which form the 20 facets of the icosahedron. The 12 vertices are composed of the penton complex, which comprises the pentameric penton base and the externally projecting trimeric fiber. The fiber has a shaft of variable length with a terminal globular knob or head domain. In most cell types in vitro, the fiber head and penton base mediate attachment and entry, respectively. Crystal structures are available of the HAd2 and HAd5 hexons (4, 5); the HAd2 penton base (6); the fiber head domains of HAd2, HAd3, HAd5, HAd12, HAd37, HAd41 (short); and a construct of HAd2 fiber head plus part of its shaft (7-13) (reviewed in Ref. 14). In addition, there are recent high resolution cryoelectron microscopy reconstructions of the entire virus (15, 16). Based on similarities in overall capsid architecture and capsid protein structure, Adenoviridae probably share an ancient common ancestor with bacteriophages of the Tectiviridae family (17-19).The initial stages of adenoviral infection, cellular attachment, ...

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confidence: 97%

Structural and Mutational Analysis of Human Ad37 and Canine Adenovirus 2 Fiber Heads in Complex with the D1 Domain of Coxsackie and Adenovirus Receptor

Seiradake

Lortat‐Jacob

Billet

et al. 2006

Journal of Biological Chemistry

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“…9,10 Ad5 vectors with the S408E mutation are unable to directly interact with CAR. 9,11 The deletion of the RGD (Arg-Gly-Asp) motif within the penton base was shown to affect not only binding to a v integrin and the subsequent internalization 12 but also the escape of viral particles from the endosomes. 13 Whereas the infection efficiency of Ad5 vectors with the S408E mutation or RGD deletion in cell culture was severely reduced, the liver tropism of these vectors in vivo remained unchanged.…”

Section: Introductionmentioning

confidence: 99%

“…13 Whereas the infection efficiency of Ad5 vectors with the S408E mutation or RGD deletion in cell culture was severely reduced, the liver tropism of these vectors in vivo remained unchanged. 9, 10 Smith et al 14 have described the putative HSPG ablating mutation S* with the Lys-Lys-Thr-Lys (KKTK) motif in the fiber shaft being changed to Gly-Ala-Gly-Ala (GAGA). Reduced gene transfer with the resulting vector was not only seen in vitro but also in vivo with 15-fold reduced liver transduction after systemic application.…”

Section: Introductionmentioning

confidence: 99%

“…To interfere with CAR binding, Ad5 vectors were developed with modifications in the AB loop of the fiber knob, for example S408E. 9,10 Ad5 vectors with the S408E mutation are unable to directly interact with CAR. 9,11 The deletion of the RGD (Arg-Gly-Asp) motif within the penton base was shown to affect not only binding to a v integrin and the subsequent internalization 12 but also the escape of viral particles from the endosomes.…”

Section: Introductionmentioning

confidence: 99%

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Targeting of adenovirus vectors carrying a tumor cell-specific peptide: in vitro and in vivo studies

et al. 2007

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Previously, we have identified a tumor cell-specific peptide, HEW, by panning of phage display libraries on the human colorectal cancer cell line WiDr. In this report we demonstrate that this peptide can modify the infection properties of adenovirus vectors. Increased infectivity of replication-deficient adenovirus 5 vectors in WiDr cells was observed upon genetic insertion of the HEW peptide in the HI loop of the fiber knob. Moreover, whereas the coxsackie and adenovirus receptor (CAR)-ablating fiber mutation S408E abolished apparent infection in CAR-positive WiDr cells, the insertion of HEW completely restored infectivity toward these cells in vitro. To assess whether the de-and re-targeted infection profile was maintained in vivo, the fiber-modified adenovirus vectors were injected intratumorally or intravenously in WiDr tumor-bearing Swiss nu/nu mice. No significant differences in efficiency of infection could be observed suggesting alternative viral uptake mechanisms in vivo. Next, we have included the fiber shaft mutation S* in our studies, which was described to confer a de-targeted phenotype in vivo. Reduced gene transfer due to the S* mutation both in vitro and in vivo could be confirmed. Insertion of HEW in the HI knob loop of shaft-mutated fiber, however, did not rescue infectivity in target cells neither in vitro nor in vivo. We demonstrate the efficient ligand-mediated re-targeting of adenoviral vector infection to the human cancer cell line WiDr. The lack of apparent re-targeting in the in vivo situation is described.

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“…Some studies have demonstrated that ablation of CAR binding does not alter the biodistribution or toxicity of systemically administered adenovirus in mice. 10,11 In both studies, the biodistribution was monitored by Q -PCR and transgene expression assays. A more recent report has demonstrated that a CAR -ablated vector with the KO1 mutation in the AB loop gave higher levels of hepatic gene transduction than vectors containing wild -type fiber.…”

Section: Virus Retargeting Strategiesmentioning

confidence: 99%

Adenoviral vectors: Systemic delivery and tumor targeting

Green

Seymour

2002

Cancer Gene Ther

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Influence of adenoviral fiber mutations on viral encapsidation, infectivity and in vivo tropism

Cited by 90 publications

References 46 publications

Structural and Mutational Analysis of Human Ad37 and Canine Adenovirus 2 Fiber Heads in Complex with the D1 Domain of Coxsackie and Adenovirus Receptor

Structural and Mutational Analysis of Human Ad37 and Canine Adenovirus 2 Fiber Heads in Complex with the D1 Domain of Coxsackie and Adenovirus Receptor

Targeting of adenovirus vectors carrying a tumor cell-specific peptide: in vitro and in vivo studies

Adenoviral vectors: Systemic delivery and tumor targeting

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