Enhanced suicide gene effect by adenoviral transduction of a VP22-cytosine deaminase (CD) fusion gene

Wybranietz, W. A.; Groß, Caspar; Phelan, Anne M.; O’Hare, Peter; Spiegel, Martin; Graepler, Florian; Bitzer, Michael; Stähler, P; Gregor, Michael; Lauer, Ulrich M.

doi:10.1038/sj.gt.3301564

Cited by 54 publications

(53 citation statements)

References 50 publications

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“…39 This property of VP22 was first demonstrated with the protein alone or fused to GFP, and it was then extended to other fusion partners that have a therapeutic potential in gene therapy. 37,38,[40][41][42][43] Using this very attractive strategy, an increase in the BE was reported with TK/ VP22 versus TK alone; however, the cytotoxic effect enhancement was very limited, 37,38 and it was only detected when a high percentage of cells expressed the fusion protein. 37 Based on these results, we hypothesized that TK30 fused to VP22 should increase the BE independently of the level of connexins.…”

Section: Introductionmentioning

confidence: 99%

Direct evidence for the absence of intercellular trafficking of VP22 fused to GFP or to the herpes simplex virus thymidine kinase

et al. 2004

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The treatment of solid tumors by retroviral delivery of the herpes simplex virus thymidine kinase (TK) followed by ganciclovir (GCV) treatment has so far shown only limited success in patients. One major drawback in this approach is the lack of efficient in vivo gene delivery to cancer cells. Although, the transduction of every single tumor cell is not a requirement since the bystander effect (BE) mediated by gap junctions allows the diffusion of the toxic GCV metabolites from TK-expressing cells toward untransduced cells. To render the TK/GCV approach more potent, and independent of the level of gap junctions, we have tested the efficiency of a TK mutant (TK30) fused to VP22, a herpes simplex protein that seems to be capable of intercellular trafficking. We failed to detect an increase in the BE with cells expressing VP22 fused to TK30 versus cells containing TK30 alone, and this result forced us to reinvestigate the trafficking properties of VP22. Using very sensitive Western blot and fluorescence assays, we were not able to detect the spread of VP22 fused either to TK30 or GFP. These results indicate that VP22 cannot be used as a cargo to translocate TK30 or GFP.

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Section: Introductionmentioning

confidence: 99%

Direct evidence for the absence of intercellular trafficking of VP22 fused to GFP or to the herpes simplex virus thymidine kinase

et al. 2004

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“…We already observed this phenomenon with a P21-VP22 and a P53-VP22 fusion (unpublished work), and other groups also described this pattern when working with N-terminal fusions. 8,10,12 Usually, these experiments reveal a cleavage product at a size slightly superior to the size of the protein fused to VP22. Another cleavage can also appear at the junction between VP22 and its partner.…”

Section: Antitumour Activity Of P27vp22mentioning

confidence: 95%

“…This was recently confirmed in a study using a VP22-CD (cytosine deaminase) protein. 12 This fusion was not able to spread, whereas a CD-VP22 fusion showed efficient intercellular transport. The second persistent problem remains the cleavage of fusion proteins.…”

Section: Antitumour Activity Of P27vp22mentioning

confidence: 96%

“…[3][4][5][6][7] Thus, linkage to VP22 has permitted to enhance the biological activity of several proteins such as the tumour suppressor p53 8,9 or the products of the suicide genes thymidine kinase 10,11 and cytosine deaminase. 12 It was also used to deliver functional Flp recombinase 13 and to carry SV40 large T antigen to induce cell cycle reentry in terminally differentiated muscle cells. 14 As a result of its spreading property, VP22 has also been used as an adjuvant in DNA vaccination assays.…”

Section: Introductionmentioning

confidence: 99%

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Intercellular trafficking and enhanced in vivo antitumour activity of a non-virally delivered P27-VP22 fusion protein

Zavaglia

Eymin

et al. 2003

Gene Ther

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VP22, a structural protein from herpes simplex virus type I, exhibits the unique property of intercellular trafficking. This protein is exported from primary expressing cells and subsequently imported into neighbouring cells. This property is conserved when VP22 is genetically fused to a protein, making it a promising tool to enhance the delivery of a gene product. We chose to study the intercellular transport and biological effect of a fusion protein between the putative tumour suppressor gene p27 Kip1 and VP22. We show that in vitro, P27VP22 is able to spread as efficiently as VP22. Functionality of the P27VP22 protein was demonstrated by its ability to inhibit cyclin/CDK2 complexes activity. In proliferation and clonogenicity assays, transfection with the P27VP22 plasmid resulted in a stronger cell growth inhibition when compared to transfection with the p27 Kip1 vector. In vivo, sub cutaneous tumours established in nude mice were injected with naked DNA encoding P27 or P27VP22. Our results show that P27VP22 can spread in vivo and that injections of the P27VP22 plasmid resulted in a significantly greater antitumour activity than injections of the P27 plasmid. This study confirms the usefulness of VP22-mediated delivery and suggests that P27VP22 may have applications in cancer gene therapy.

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“…Most reports have concentrated on anti-tumor therapies either by induction of apoptosis upon delivery of functional p53 fused to VP22 (Phelan et al, 1998;Wills et al, 2001;Zender et al, 2002) or by delivering enzymes (thymidine kinase or cytosine deaminase) that allow the conversion of commonly used prodrugs into toxic compounds thereby inducing cell death (Dilber et al, 1999;Liu et al, 2001;Wybranietz et al, 2001). In some cases, coculture of VP22-producing transfected cells and recipient cells was used.…”

Section: Vp22mentioning

confidence: 99%

Protein Transduction: A Novel Tool for Tissue Regeneration

Cardoso

Leonhardt²

2002

Biological Chemistry

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Tissue regeneration in humans is limited and excludes vitals organs like heart and brain. Transformation experiments with oncogenes like T antigen have shown that retrodifferentiation of the respective cells is possible but hard to control. To bypass the risk of cancer formation a protein therapy approach has been developed. The transient delivery of proteins rather than genes could still induce terminallydifferentiated cells to reenter the cell cycle. This approach takes advantage of proteintransducing domains that mediate the transfer of cargo proteins into cells. The goal of this brief review is to outline the basics of protein transduction and to discuss potential applications for tissue regeneration.

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Enhanced suicide gene effect by adenoviral transduction of a VP22-cytosine deaminase (CD) fusion gene

Abstract: The low transduction efficiency of viral and nonviral vectors is a major limitation in tumour gene

Cited by 54 publications

References 50 publications

Direct evidence for the absence of intercellular trafficking of VP22 fused to GFP or to the herpes simplex virus thymidine kinase

Direct evidence for the absence of intercellular trafficking of VP22 fused to GFP or to the herpes simplex virus thymidine kinase

Intercellular trafficking and enhanced in vivo antitumour activity of a non-virally delivered P27-VP22 fusion protein

Protein Transduction: A Novel Tool for Tissue Regeneration

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