Modular adeno-associated virus (rAAV) vectors used for cellular virus-directed enzyme prodrug therapy

Hagen, Sven; Baumann, Tobias; Wagner, Hanna J.; Morath, Volker; Kaufmann, Beate; Fischer, Adrian; Bergmann, Stefan; Schindler, Patrick; Arndt, Katja M.; Müller, Kristian M.

doi:10.1038/srep03759

Cited by 32 publications

(31 citation statements)

References 53 publications

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“…The presence of positively charged residues as those of the His-tag at this position can reconstitute HSPG binding and natural receptor usage for cell entry 34,35 . Owing to these potential risks and the absence of a functional proof that a His-tag in position 587 indeed efficiently depletes DARPin-deficient particles 30 , we decided to place the His-tag N-terminally of the DARPin, thus avoiding both of the above mentioned risks. This step enhanced in addition gene delivery more than 10-fold.…”

Section: Discussionmentioning

confidence: 99%

“…AAV particles have been previously modified with Histags to facilitate particle purification by HisTrap columns 28,29 . Moreover, position 587 of the AAV-2 capsid, previously identified as possible insertion site for His-tags 29 , has recently been exploited to separate DARPin-displaying and DARPindeficient particles 30 . However, since His-tag and DARPin are not physically linked in this setting, recombination, which is a frequent event in packaging cells 31 , can give rise to DARPindeficient but His-tag-displaying particles.…”

Section: Discussionmentioning

confidence: 99%

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Off-target-free gene delivery by affinity-purified receptor-targeted viral vectors

et al. 2015

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We describe receptor-targeted adeno-associated viral (AAV) vectors that allow genetic modification of rare cell types ex vivo and in vivo while showing no detectable off-targeting. Displaying designed ankyrin repeat proteins (DARPins) on the viral capsid and carefully depleting DARPin-deficient particles, AAV vectors were made specific for Her2/neu, EpCAM or CD4. A single intravenous administration of vector targeted to the tumour antigen Her2/neu was sufficient to track 75% of all tumour sites and to extend survival longer than the cytostatic antibody Herceptin. CD4-targeted AAVs hit human CD4-positive cells present in spleen of a humanized mouse model, while CD8-positive cells as well as liver or other off-target organs remained unmodified. Mimicking conditions of circulating tumour cells, EpCAM-AAV detected single tumour cells in human blood opening the avenue for tumour stem cell tracking. Thus, the approach developed here delivers genes to target cell types of choice with antibody-like specificity.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Off-target-free gene delivery by affinity-purified receptor-targeted viral vectors

et al. 2015

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“…Viral particles containing a gene therapy vector encoding HSV-TK, in combination with ganciclovir, effectively suppressed xenograft tumor growth, without causing hepatotoxicity [93]. Similar viral particles were obtained using EGFR-specific DARPin and affibody, and both agents showed selective toxicity towards EGFR-positive cells in vitro [94].…”

Section: Application Of Darpins In Designing Oncolytic Virusesmentioning

confidence: 79%

DARPins: Promising Scaffolds for Theranostics

Shilova

Deyev

2019

Acta Naturae

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Monoclonal antibodies are the classical basis for targeted therapy, but the development of alternative binding proteins has made it possible to use non-immunoglobulin proteins as targeting modules. The advantages of DARPins, scaffold proteins based on ankyrin repeats, over antibodies are as follows: small size, stability over a wide range of temperatures and pH values, low aggregation tendency, and ease of production in heterologous expression systems. The differences in the structure of the paratope of DARPin and antibodies broaden the spectrum of target molecules, while the ease of creating hybrid fusion proteins allows one to obtain bispecific and multivalent constructs. In this article, we summarize recent data on the development of therapeutic and imaging compounds based on DARPins.

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“…The evaluation of cytotoxic activity has been done by assessing the stage of cell death as early-and late-apoptotic or necrotic cells. 15,18,19 Results and discussion…”

Section: Introductionmentioning

confidence: 99%

Virus-directed enzyme prodrug therapy and the assessment of the cytotoxic impact of some benzimidazole derivatives

et al. 2017

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Virus-directed enzyme prodrug therapy is one of the major strategy of increasing cytotoxicity of bioreductive agents. This research intended to examine new selected benzimidazole derivatives as a substrate for nitroreductase, the enzyme involved in nitroreduction which is responsible to the production of cytotoxic metabolites. In this way, the selectivity and strength of cytotoxicity can be raised. The effect of benzimidazoles on virus transfected cells and non-virus transfected cells A549 cell line was established by Annexin V + propidium iodide test, western blot, and polymerase chain reaction analysis of specific pro- and anti-apoptotic proteins in the corresponding gene expression and additionally nitroreductase gene expression. Our results proved the pro-apoptotic properties of all tested compounds in normoxia and hypoxia, especially according to virused A549 cells where the time of exposition was reduced from 48 to 4 h. In this shorten period of time, the strongest activity was shown by N-oxide compounds with nitro-groups. The apoptosis was confirmed by generation of BAX gene and protein and reduction of BCL2 gene and protein.

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Modular adeno-associated virus (rAAV) vectors used for cellular virus-directed enzyme prodrug therapy

Cited by 32 publications

References 53 publications

Off-target-free gene delivery by affinity-purified receptor-targeted viral vectors

Off-target-free gene delivery by affinity-purified receptor-targeted viral vectors

DARPins: Promising Scaffolds for Theranostics

Virus-directed enzyme prodrug therapy and the assessment of the cytotoxic impact of some benzimidazole derivatives

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