title: NF-κB promotes leaky expression of adenovirus genes in a replication-incompetent adenovirus vector.
abstract: The replication-incompetent adenovirus (Ad) vector is one of the most promising vectors for gene therapy; however, systemic administration of Ad vectors results in severe hepatotoxicities, partly due to the leaky expression of Ad genes in the liver. Here we show that nuclear factor-kappa B (NF-κB) mediates the leaky expression of Ad genes from the Ad vector genome, and that the inhibition of NF-κB leads to the suppression of Ad gene expression and hepatotoxicities following transduction with Ad vectors. Activation of NF-κB by recombinant tumor necrosis factor (TNF)-α significantly enhanced the leaky expression of Ad genes. More than 50% suppression of the Ad gene expression was found by inhibitors of NF-κB signaling and siRNA-mediated knockdown of NF-κB. Similar results were found when cells were infected with wild-type Ad. Compared with a conventional Ad vector, an Ad vector expressing a dominant-negative IκBα (Adv-CADNIκBα), which is a negative regulator of NF-κB, mediated approximately 70% suppression of the leaky expression of Ad genes in the liver. Adv-CADNIκBα did not induce apparent hepatotoxicities. These results indicate that inhibition of NF-κB leads to suppression of Ad vector-mediated tissue damages via not only suppression of inflammatory responses but also reduction in the leaky expression of Ad genes.
authors.affiliation: Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Machitani
authors.affiliation: Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan.|Laboratory of Regulatory Sciences for Oligonucleotide Therapeutics, Clinical Drug Development Unit, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka, 565-0871, Japan.
authors.forename: F
authors.initials: F
authors.lastname: Sakurai
authors.affiliation: Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Wakabayashi
authors.affiliation: Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Nakatani
authors.affiliation: Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan.|Faculty of Pharmacy, Osaka Ohtani University, 3-11-1 Nishikiorikita, Tondabayashi, Osaka, 584-8540, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Shimizu
authors.affiliation: Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Tachibana
authors.affiliation: Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan.|Laboratory of Hepatocyte Regulation, National Institutes of Biomedical Innovation, Health and Nutrition, 7-6-8 Saito, Asagi, Ibaraki, Osaka 567-0085, Japan.|iPS Cell-Based Research Project on Hepatic Toxicity and Metabolism, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan.|Global Center for Advanced Medical Engineering and Informatics, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan.|Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Mizuguchi
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D017502:Adenovirus E2 Proteins / Q000235:genetics; D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D005260:Female; D015967:Gene Expression Regulation, Viral / Q000187:drug effects; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D016898:Interferon-alpha / Q000494:pharmacology; D008099:Liver / Q000378:metabolism / Q000821:virology; D051379:Mice; D018345:Mice, Knockout; D016328:NF-kappa B / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D017384:Sequence Deletion; D015533:Transcriptional Activation; D014779:Virus Replication / Q000187:drug effects
chemical_list: D017502:Adenovirus E2 Proteins; D016898:Interferon-alpha; D016328:NF-kappa B
references.citation: Mol Genet Metab. 2005 Mar;84(3):278-88
references.pmid: 15694178
references.citation: J Clin Invest. 2005 Oct;115(10):2625-32
references.pmid: 16200195
references.citation: Gene Ther. 2005 Oct;12(19):1424-33
references.pmid: 15944730
references.citation: J Virol. 1996 Oct;70(10):7030-8
references.pmid: 8794347
references.citation: J Clin Invest. 2001 Jan;107(2):143-51
references.pmid: 11160127
references.citation: J Virol. 1996 Dec;70(12):8934-43
references.pmid: 8971023
references.citation: Mol Ther. 2001 May;3(5 Pt 1):697-707
references.pmid: 11356075
references.citation: J Virol. 2004 Dec;78(24):13582-90
references.pmid: 15564469
references.citation: J Virol. 1998 Nov;72(11):9267-77
references.pmid: 9765474
references.citation: J Virol. 1997 Jun;71(6):4626-37
references.pmid: 9151856
references.citation: Virus Res. 2013 Dec 26;178(2):357-63
references.pmid: 24055658
references.citation: J Clin Invest. 1998 Feb 15;101(4):802-11
references.pmid: 9466975
references.citation: J Cell Biol. 1996 Feb;132(4):511-22
references.pmid: 8647884
references.citation: Virology. 1998 Aug 1;247(2):212-22
references.pmid: 9705914
references.citation: J Virol. 1998 Mar;72(3):2022-32
references.pmid: 9499056
references.citation: J Virol. 1997 Jun;71(6):4638-48
references.pmid: 9151857
references.citation: Mol Ther Methods Clin Dev. 2014 Sep 03;1:14035
references.pmid: 26015975
references.citation: J Gene Med. 2004 Feb;6 Suppl 1:S164-71
references.pmid: 14978759
references.citation: Mol Ther. 2005 Sep;12(3):547-54
references.pmid: 15950541
references.citation: Hepatology. 1997 Oct;26(4):949-56
references.pmid: 9328318
references.citation: Cell. 1992 Mar 6;68(5):855-67
references.pmid: 1547487
references.citation: J Immunol. 1989 Dec 1;143(11):3806-12
references.pmid: 2531188
references.citation: Biomaterials. 2013 May;34(16):4191-201
references.pmid: 23473963
references.citation: Hum Gene Ther. 2004 Nov;15(11):1022-33
references.pmid: 15610603
references.citation: Virology. 1995 Aug 1;211(1):251-60
references.pmid: 7645218
references.citation: Mol Pharm. 2011 Aug 1;8(4):1430-5
references.pmid: 21682288
references.citation: Gene Ther. 2003 Jun;10(11):991-8
references.pmid: 12756420
references.citation: Int J Pharm. 2008 Apr 16;354(1-2):9-15
references.pmid: 17640834
references.citation: Hum Gene Ther. 1999 Aug 10;10(12):2013-7
references.pmid: 10466635
references.citation: Immunity. 1995 Mar;2(3):223-38
references.pmid: 7697543
references.citation: Mol Ther. 2002 Jan;5(1):63-73
references.pmid: 11786047
references.citation: Proc Natl Acad Sci U S A. 1994 May 10;91(10):4407-11
references.pmid: 8183921
references.citation: Hum Gene Ther. 1998 Nov 20;9(17):2577-83
references.pmid: 9853524
references.citation: Proc Natl Acad Sci U S A. 1994 Jun 21;91(13):6196-200
references.pmid: 8016137
references.citation: Mol Ther. 2008 Oct;16(10):1719-26
references.pmid: 18665157
references.citation: J Control Release. 2011 Sep 25;154(3):285-9
references.pmid: 21703313