title: Validation of IKK beta as therapeutic target in airway inflammatory disease by adenoviral-mediated delivery of dominant-negative IKK beta to pulmonary epithelial cells.
abstract: Asthma is an inflammatory disease of the lungs and the transcription factor NF-kappa B regulates the production of numerous inflammatory mediators that may have a role in the pathogenesis of asthma. Hence, the signalling pathways leading to NF-kappa B activation are considered prime targets for novel anti-inflammatory therapies. The prevention of NF-kappa B activity in mice, through the knockout of IKK beta or p65, causes fatal liver degeneration in utero making it difficult to determine the full implications of inhibiting NF-kappaB activity in tissues physiologically relevant to human diseases. This study used adenovirus delivery of a dominant inhibitor of NF-kappaB (I kappa B alpha delta N) and dominant-negative IKK alpha (IKK alpha(KM)) and IKK beta (IKK beta(KA)) to investigate the role of the individual IKKs in NF-kappa B activation and inflammatory gene transcription by human pulmonary A549 cells. Overexpression of IKK beta(KA) or I kappa B alpha delta N prevented NF-kappa B-dependent transcription and DNA binding. IKK beta(KA) also prevented I kappa B alpha kinase activity. Similarly, IKK beta(KA) and I kappa B alpha delta N overexpression also inhibited IL-1beta- and TNF alpha-dependent increases in ICAM-1, IL-8 and GM-CSF in addition to IL-1beta-mediated increases in cyclooxygenase-2 expression, whereas IKK alpha(KM) overexpression had little effect on these outputs. IKK beta(KA) also reduced cell viability and induced caspase-3 and PARP cleavage regardless of the stimuli, indicating the induction of apoptosis. This effect seemed to be directly related to IKK beta kinase activity since I kappa B alpha delta N only induced PARP cleavage in TNF alpha-treated cells. These results demonstrate that inhibition of IKK beta and NF-kappa B suppresses inflammatory mediator production and reduces A549 cell viability. Thus, novel therapies that target IKK beta could have potent anti-inflammatory effects and may be beneficial in the treatment of certain cancers.
authors.affiliation: Department of Thoracic Medicine, National Heart and Lung Institute, Imperial College, Faculty of Medicine, London.
authors.forename: Matthew C
authors.initials: MC
authors.lastname: Catley
authors.forename: Joanna E
authors.initials: JE
authors.lastname: Chivers
authors.forename: Neil S
authors.initials: NS
authors.lastname: Holden
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Barnes
authors.forename: Robert
authors.initials: R
authors.lastname: Newton
mesh_terms: D000256:Adenoviridae; D017209:Apoptosis; D001249:Asthma / Q000378:metabolism; D002460:Cell Line; D004847:Epithelial Cells / Q000378:metabolism; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D051550:I-kappa B Kinase; D016328:NF-kappa B / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000502:physiology*; D020545:Respiratory Mucosa / Q000166:cytology / Q000378:metabolism*; D014158:Transcription, Genetic / Q000187:drug effects / Q000502:physiology
chemical_list: D016328:NF-kappa B; D017346:Protein Serine-Threonine Kinases; C496559:CHUK protein, human; D051550:I-kappa B Kinase; C496560:IKBKB protein, human; C496565:IKBKE protein, human
references.citation: Nature. 1999 Nov 25;402(6760 Suppl):B31-8
references.pmid: 10586893
references.citation: J Biol Chem. 1999 Jul 9;274(28):19965-72
references.pmid: 10391945
references.citation: Am J Respir Crit Care Med. 2000 May;161(5):1720-45
references.pmid: 10806180
references.citation: J Exp Med. 2001 Mar 5;193(5):631-6
references.pmid: 11238593
references.citation: Immunity. 2001 Mar;14(3):217-30
references.pmid: 11290332
references.citation: Trends Cell Biol. 2001 Jul;11(7):304-11
references.pmid: 11413042
references.citation: Mol Cell. 2001 Nov;8(5):1005-16
references.pmid: 11741536
references.citation: Nat Immunol. 2002 Mar;3(3):221-7
references.pmid: 11875461
references.citation: Am J Respir Crit Care Med. 2002 Apr 1;165(7):996-1004
references.pmid: 11934728
references.citation: Cell. 2002 Apr;109 Suppl:S81-96
references.pmid: 11983155
references.citation: Nature. 2002 Jun 20;417(6891):861-6
references.pmid: 12075355
references.citation: Apoptosis. 2002 Aug;7(4):321-8
references.pmid: 12101391
references.citation: Trends Mol Med. 2002 Aug;8(8):385-9
references.pmid: 12127724
references.citation: J Biol Chem. 2002 Nov 22;277(47):45129-40
references.pmid: 12221085
references.citation: Nature. 2003 Jun 5;423(6940):655-9
references.pmid: 12789342
references.citation: Nature. 2003 Jun 5;423(6940):659-63
references.pmid: 12789343
references.citation: FEBS Lett. 2003 Jul 17;547(1-3):75-9
references.pmid: 12860389
references.citation: J Biol Chem. 1994 Dec 9;269(49):30761-4
references.pmid: 7983002
references.citation: Br J Pharmacol. 1994 Nov;113(3):1008-14
references.pmid: 7858842
references.citation: Biochem Biophys Res Commun. 1996 Jan 17;218(2):518-23
references.pmid: 8561789
references.citation: Proc Natl Acad Sci U S A. 1996 Jul 23;93(15):7464-9
references.pmid: 8755496
references.citation: EMBO J. 1996 Dec 2;15(23):6716-26
references.pmid: 8978697
references.citation: Cell. 1997 Oct 17;91(2):243-52
references.pmid: 9346241
references.citation: Science. 1997 Oct 31;278(5339):860-6
references.pmid: 9346484
references.citation: J Biol Chem. 1998 Dec 11;273(50):33533-9
references.pmid: 9837934
references.citation: J Biol Chem. 1999 Jan 15;274(3):1715-22
references.pmid: 9880552
references.citation: Science. 1999 Apr 9;284(5412):321-5
references.pmid: 10195897
references.citation: J Exp Med. 1999 Jun 7;189(11):1839-45
references.pmid: 10359587
references.citation: Am J Respir Cell Mol Biol. 2000 May;22(5):582-9
references.pmid: 10783130