Functional and Selective Targeting of Adenovirus to High-Affinity Fcγ Receptor I-Positive Cells by Using a Bispecific Hybrid Adapter

Ebbinghaus, Christina; Al-Jaibaji, Ahmed; Operschall, Elisabeth; Schöffel, A.; Peter, Isabelle S.; Greber, Urs F.; Hemmi, Silvio

doi:10.1128/jvi.75.1.480-489.2001

Cited by 68 publications

(60 citation statements)

References 69 publications

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“…29,63 For detection of apoptotic/ necrotic cells, cultured cells were grown in six-well plates to subconfluency and infected with different Ads at MOI 20. The cells were analysed for cell viability by annexin/propidium iodide staining at 24, 40 and 65 h p.i.…”

Section: Flow Cytometrymentioning

confidence: 99%

Melanoma cultures show different susceptibility towards E1A-, E1B-19 kDa- and fiber-modified replication-competent adenoviruses

et al. 2006

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Replicating adenovirus (Ad) vectors with tumour tissue specificity hold great promise for treatment of cancer. We have recently constructed a conditionally replicating Ad5 AdDEP-TETP inducing tumour regression in a xenograft mouse model. For further improvement of this vector, we introduced four genetic modifications and analysed the viral cytotoxicity in a large panel of melanoma cell lines and patient-derived melanoma cells. (1) The antiapoptotic gene E1B-19 kDa (D19 mutant) was deleted increasing the cytolytic activity in 18 of 21 melanoma cells. (2) Introduction of the E1A 122-129 deletion (D24 mutant), suggested to attenuate viral replication in cell cycle-arrested cells, did not abrogate this activity and increased the cytolytic activity in two of 21 melanoma cells. (3) We inserted an RGD sequence into the fiber to extend viral tropism to av integrinexpressing cells, and (4) swapped the fiber with the Ad35 fiber (F35) enhancing the tropism to malignant melanoma cells expressing CD46. The RGD-fiber modification strongly increased cytolysis in all of the 11 CAR-low melanoma cells. The F35 fiber-chimeric vector boosted the cytotoxicity in nine of 11 cells. Our results show that rational engineering additively enhances the cytolytic potential of Ad vectors, a prerequisite for the development of patient-customized viral therapies. Gene Therapy (2006) 13, 893-905.

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Section: Flow Cytometrymentioning

confidence: 99%

Melanoma cultures show different susceptibility towards E1A-, E1B-19 kDa- and fiber-modified replication-competent adenoviruses

et al. 2006

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“…Previously we and others have reported the use of bispecific adapters binding to a virus capsid protein for specific adenovirus-mediated targeting (14,(26)(27)(28)(29)(30)(31)(32)(33)(34)(35)(36)(37). The successful execution of the adapter strategy depends critically on our ability to design protein adapters for Ad that allow the targeting of any desired disease-associated receptor and whose association with the Ad virion is highly stable and can be fine-tuned to be compatible with the virion's structural integrity and its stepwise disintegration during cell entry (38).…”

mentioning

confidence: 99%

Development of a generic adenovirus delivery system based on structure-guided design of bispecific trimeric DARPin adapters

Dreier

Honegger

Hess

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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Adenoviruses (Ads) have shown promise as vectors for gene delivery in clinical trials. Efficient viral targeting to a tissue of choice requires both ablation of the virus' original tropism and engineering of an efficient receptor-mediated uptake by a specific cell population. We have developed a series of adapters binding to the virus with such high affinity that they remain fully bound for >10 d, block its natural receptor binding site and mediate interaction with a surface receptor of choice. The adapter contains two fused modules, both consisting of designed ankyrin repeat proteins (DARPins), one binding to the fiber knob of adenovirus serotype 5 and the other binding to various tumor markers. By solving the crystal structure of the complex of the trimeric knob with three bound DARPins at 1.95-Å resolution, we could use computer modeling to design a link to a trimeric protein of extraordinary kinetic stability, the capsid protein SHP from the lambdoid phage 21. We arrived at a module which binds the knob like a trimeric clamp. When this clamp was fused with DARPins of varying specificities, it enabled adenovirus serotype 5-mediated delivery of a transgene in a human epidermal growth factor receptor 2-, epidermal growth factor receptor-, or epithelial cell adhesion molecule-dependent manner with transduction efficiencies comparable to or even exceeding those of Ad itself. With these adapters, efficiently produced in Escherichia coli, Ad can be converted rapidly to new receptor specificities using any ligand as the receptor-binding moiety. Prefabricated Ads with different payloads thus can be retargeted readily to many cell types of choice.protein design | tumor targeting | viral retargeting | X-ray crystallography | protein engineering

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“…Successful retargeting to new receptors rely, however, on precise activation of the coreceptors, exemplified by the low infectivity of Adv retargeted to the transferrin receptor, while delivery to CD64 in hematopoietic cells or to integrins (by means of diabodies or insertion of the RGD sequence in the H1 loop of the fiber-knob) was more successful. 16,36 CAR seems not to be crucial for photochemical transduction either. The transgene expression from the fiber-manipulated virus AdRGD-GFP 16 was enhanced after photochemical treatment, showing that photochemical transduction also can be effective in combination with retargeted viral particles.…”

Section: Discussionmentioning

confidence: 91%

PCI-enhanced adenoviral transduction employs the known uptake mechanism of adenoviral particles

et al. 2005

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The development of methods for efficient and specific delivery of therapeutic genes into target tissues is an important issue for further development of in vivo gene therapy. In the present study, the physical targeting technique, photochemical internalization (PCI), has been used together with adenovirus. The combination of PCI and adenoviral transduction has previously been shown to be favorable compared to adenovirus used alone, and the aim of this study was to verify the role of the adenoviral receptors and identify the uptake pathway used by adenoviral particles in photochemically treated cells. All examined cell lines showed augmented transduction efficiency after PCI-treatment, with a maximum of 13-fold increase in transgene expression compared to conventionally infected cells. Blocking of CAR induced a complete inhibition of PCI-enhanced transgene expression. However, photochemical treatment managed to enhance the transduction efficiency of the retargeted virus AdRGD-GFP showing also that the virus-CAR interaction is not vital for obtaining a photochemical effect on adenoviral transduction. Blocking the a V -integrins reduced the gene expression significantly in photochemically treated cells. Subjecting HeLa cells expressing negative mutant-dynamin to light treatment after infection gave no significant increase in gene transfer, while the gene transfer were enhanced seven-fold in cells with wild-type dynamin. Furthermore, chlorpromazine inhibited photochemical transduction in a dose-dependent manner, whereas Filipin III had no effect on the gene transfer. In summary, the data presented imply that adenoviral receptor binding is important and clathrin-mediated endocytosis is the predominant uptake mechanism for adenoviral particles in photochemically treated cells.

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Functional and Selective Targeting of Adenovirus to High-Affinity Fcγ Receptor I-Positive Cells by Using a Bispecific Hybrid Adapter

Cited by 68 publications

References 69 publications

Melanoma cultures show different susceptibility towards E1A-, E1B-19 kDa- and fiber-modified replication-competent adenoviruses

Melanoma cultures show different susceptibility towards E1A-, E1B-19 kDa- and fiber-modified replication-competent adenoviruses

Development of a generic adenovirus delivery system based on structure-guided design of bispecific trimeric DARPin adapters

PCI-enhanced adenoviral transduction employs the known uptake mechanism of adenoviral particles

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