Activation of p38 and ERK signaling during adenovirus vector cell entry lead to expression of the C-X-C chemokine IP-10.
The use of adenovirus vectors for human gene therapy is limited by potent inflammatory responses that result in significant morbidity. In kidney-derived epithelial cells (REC), activation of extracellular signal-regulated kinase 1/2 (ERK) and p38 kinase (p38) pathways occurred within 20 min of transduction with the serotype 5 adenovirus vector AdCMV beta gal. Inhibition of ERK and p38 with U0126 and SB203580, respectively, reduced the expression of IP-10 mRNA following transduction with AdCMV beta gal. To determine the role of the coxsackievirus-adenovirus receptor (CAR) or alpha(v) integrins in the activation of ERK and p38 and the expression of IP-10, REC cells were transduced with the fiber-modified and RGD-deleted adenovirus vectors AdL.F(RAEK-HA) and AdL.PB(HA), respectively. Compared with the wild-type capsid vector Ad5Luc, transduction with AdL.F(RAEK-HA) and AdL.PB(HA) resulted in reduced ERK-p38 activation and less IP-10 mRNA expression. The decreased IP-10 expression induced by the tropism-modified vectors was due to diminished transduction, since increasing multiplicity of infection resulted in increased IP-10 expression. Inhibition of adenovirus penetration with bafilomycin A1 or ammonium chloride attenuated the activation of ERK-p38 and IP-10 mRNA expression following infection, suggesting that endosomal escape was required to trigger these pathways. In vivo, direct inhibition of ERK and p38 signaling pathways inhibited adenovirus vector-induced IP-10 expression in mouse liver 1 h following transduction. These results demonstrate the importance of signaling via ERK and p38 in the early host response to adenovirus vectors and will permit the development of novel strategies to improve the safety and efficacy of these agents in human gene therapy.
Tibbles
Lee Anne
LA
Department of Medicine. Libin Gene Therapy Unit, University of Calgary Calgary, Alberta T2N 4N1, Canada.
Spurrell
Jason C L
JC
Bowen
Gloria P
GP
Liu
Qiang
Q
Lam
Mindy
M
Zaiss
Anne K
AK
Robbins
Stephen M
SM
Hollenberg
Morley D
MD
Wickham
Thomas J
TJ
Muruve
Daniel A
DA
D000258:Adenovirus Infections, Human; D000260:Adenoviruses, Human / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology; D000818:Animals; D002461:Cell Line, Transformed; D054357:Chemokine CXCL10; D019743:Chemokines, CXC / Q000235:genetics / Q000378:metabolism*; D000090702:Constitutive Androstane Receptor; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D020935:MAP Kinase Signaling System; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D018160:Receptors, Cytoplasmic and Nuclear; D011971:Receptors, Immunologic / Q000378:metabolism; D018000:Receptors, Peptide / Q000378:metabolism; D011991:Receptors, Virus / Q000378:metabolism; D014157:Transcription Factors; D014161:Transduction, Genetic; D048051:p38 Mitogen-Activated Protein Kinases
C568771:CLMP protein, human; C568772:CLMP protein, mouse; D054357:Chemokine CXCL10; D019743:Chemokines, CXC; D000090702:Constitutive Androstane Receptor; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D018160:Receptors, Cytoplasmic and Nuclear; D011971:Receptors, Immunologic; D018000:Receptors, Peptide; D011991:Receptors, Virus; D014157:Transcription Factors; C051382:arginyl-glycyl-aspartic acid directed cell adhesion receptor; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases