Abstract
Adenoviral gene expression that is repressed by p53 in nontransformed cells could provide a tumor-specific gene therapy approach for a large subset of tumors. Adenoviral infection in vivo induces stabilization of p53, which can be utilized for a strategy that includes p53-dependent expression of a transcriptional repressor and a target promoter, which is highly susceptible for transcriptional repression. Therefore, we constructed different versions of CMV-promoters (CMVgal) with binding sites for GAL4-DBD and investigated 11 GAL4-DBD fusion proteins to elucidate the most effective repressor domain to silence CMVgal activity.The transcriptional repressor GAL4-KRAB-A under control of a p53-dependent promoter facilitates strong CMVgal-mediated gene expression specifically in p53 mutant cells by a double-recombinant adenoviral vector (Ad-RGCdR). GAL4-KRAB-A mediates strong transcriptional repression of Ad-RGCdR in p53 wild-type cells, which could be further enhanced by preactivation of p53-signalling following low-dose chemotherapy prior to adenoviral infection. By utilizing p53 signalling involved in chemotherapy and adenoviral infection, more than 99% of Ad-RGCdR gene expression could be repressed in p53 wild-type cells. Controlled gene expression from CMVgal promoters by transcriptional repression utilizing functional p53 signalling thus provides a very effective tool for tumor-specific adenoviral gene therapy.
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Abbreviations
- DBD:
-
DNA-binding domain
- RGC:
-
ribosomal gene cluster
- MOI:
-
multiplicity of infection
- mt:
-
mutant
- wt:
-
wild type
- X-gal:
-
5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside
- dep.:
-
dependent
- therap.:
-
therapeutic
- Cons.:
-
consensus
- Seq.:
-
sequence
- Luc.:
-
luciferase
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This research was supported by a grant of the Wilhelm Sander Stiftung (98.046.2).
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Kühnel, F., Zender, L., Wirth, T. et al. Tumor-specific adenoviral gene therapy: transcriptional repression of gene expression by utilizing p53-signal transduction pathways. Cancer Gene Ther 11, 28–40 (2004). https://doi.org/10.1038/sj.cgt.7700632
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DOI: https://doi.org/10.1038/sj.cgt.7700632
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