Inhibition of repair of radiation-induced DNA damage enhances gene expression from replication-defective adenoviral vectors

Hingorani, M. and White, C.L. and Merron, A. and Peerlinck, I. and Gore, M.E. and Slade, A. and Scott, Simon D. and Nutting, C.M. and Pandha, H.S. and Melcher, A.A. and Vile, R.G. and Vassaux, G. and Harrington, K.J. (2008) Inhibition of repair of radiation-induced DNA damage enhances gene expression from replication-defective adenoviral vectors. Cancer Research, 68 (23). pp. 9771-9778. ISSN 0008-5472. (doi:https://doi.org/10.1158/0008-5472.CAN-08-1911) (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided)

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Official URL
http://dx.doi.org/10.1158/0008-5472.CAN-08-1911

Abstract

Radiation has been shown to up-regulate gene expression from adenoviral vectors in previous studies. In the current study, we show that radiation-induced dsDNA breaks and subsequent signaling through the mitogen-activated protein kinase (MAPK) pathway are responsible, at least in part, for this enhancement of transgene expression both in vitro and in vivo. Inhibitors of ataxia-telangiectasia-mutated, poly(ADP-ribose) polymerase-mutated, and DNA-dependent protein kinase (DNA-PK)-mediated DNA repair were shown to maintain dsDNA breaks (γH2AX foci) by fluorescence-activated cell sorting and microscopy. Inhibition of DNA repair was associated with increased green fluorescent protein (GFP) expression from a replication-defective adenoviral vector (Ad-CMV-GFP). Radiation-induced up-regulation of gene expression was abrogated by inhibitors of MAPK (PD980059 and U0126) and phosphatidylinositol 3-kinase (LY294002) but not by p38 MAPK inhibition. A reporter plasmid assay in which GFP was under the transcriptional control of artificial Egr-1 or cytomegalovirus promoters showed that the DNA repair inhibitors increased GFP expression only in the context of the Egr-1 promoter. In vivo administration of a water-soluble DNA-PK inhibitor (KU0060648) was shown to maintain luciferase expression in HCT116 xenografts after intratumoral delivery of Ad-RSV-Luc. These data have important implications for therapeutic strategies involving multimodality use of radiation, targeted drugs, and adenoviral gene delivery and provide a framework for evaluating potential advantageous combinatorial effects. ©2008 American Association for Cancer Research.

Item Type: Article
Uncontrolled keywords: 1,4 diamino 1,4 bis(2 aminophenylthio) 2,3 dicyanobutadiene, 2 (2 amino 3 methoxyphenyl)chromone, 2 morpholino 8 phenylchromone, adenovirus vector, ATM protein, DNA dependent protein kinase, double stranded DNA, early growth response factor 1, green fluorescent protein, ku 0060648, luciferase, mitogen activated protein kinase, mitogen activated protein kinase inhibitor, mitogen activated protein kinase p38, nicotinamide adenine dinucleotide adenosine diphosphate ribosyltransferase, phosphatidylinositol 3 kinase inhibitor, protein kinase inhibitor, unclassified drug, animal experiment, animal model, article, controlled study, Cytomegalovirus, DNA damage, DNA strand breakage, female, fluorescence activated cell sorter, gene expression regulation, human, human cell, in vitro study, in vivo study, microscopy, mouse, nonhuman, plasmid, priority journal, promoter region, Respiratory syncytial pneumovirus, signal transduction, transcription regulation, transgene, tumor xenograft, upregulation, viral gene delivery system, Adenoviridae, Animals, Cell Line, Tumor, DNA Damage, DNA Repair, DNA, Neoplasm, Early Growth Response Protein 1, Female, Gene Expression, Genetic Vectors, Green Fluorescent Proteins, HCT116 Cells, Humans, Luciferases, Mice, Mice, Nude, Neoplasms, Signal Transduction, Up-Regulation, Virus Replication
Subjects: Q Science
Q Science > QR Microbiology > QR355 Virology
Divisions: Faculties > Sciences > Medway School of Pharmacy
Depositing User: Simon Scott
Date Deposited: 01 Dec 2017 15:02 UTC
Last Modified: 11 Dec 2017 15:51 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/29430 (The current URI for this page, for reference purposes)
Scott, Simon D.: https://orcid.org/0000-0002-8290-0461
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