Functional antagonism of IL-1alpha induced gene expression profiles define the cAMP/PKA pathway as a unique regulator of IL-1alpha signaling networks.
Interleukin-1 (IL-1alpha) induced inflammatory and pro-fibrotic responses in human lung fibroblasts are mediated by activation of MAPK and NFkappaB pathways. The purpose of the present study was to broadly profile the activity of a variety of compounds which function as inhibitors of these key signaling pathways that may affect IL-1alpha mediated gene changes. A reference set of genes was derived from microarray analysis of IL-1alpha stimulated cells. The genes were chosen to provide a range of expression profiles which serve to represent the actions of the underlying signaling network. We show that G(s)-coupled receptor agonists have a unique pattern of activity as represented by their impact on IL-1alpha dependent gene changes. These effects were not mimicked by direct inhibitors of p38, JNK, MEK or IKK but were mimicked by forskolin and cAMP analogs. These findings indicate that cAMP/PKA serves as a point of convergence for regulation of IL-1alpha responses by multiple G(s)-coupled receptors and regulates IL-1alpha responses by a distinct mechanism that does not solely involve direct inhibition of p38, JNK, MEK or IKK. The data also point to a potentially useful paradigm wherein monitoring of a small subset of genes is sufficient to identify pathway activity of novel compounds.
Murray
David L
DL
Discovery Research and Chemoinformatics, GlaxoSmithKline, Research Triangle Park, NC 27709, USA. david.murray@roche.com
Johnson
Eric N
EN
Wang
Ping
P
Gauthier
Josee
J
Bing
Nan
N
Vanderwall
Dana
D
Xu
Zangwei
Z
Jensen
David
D
Cox
Richard F
RF
Kostura
Matthew
M
Sorensen
Scott D
SD
D000527:Alprostadil / Q000031:analogs & derivatives / Q000494:pharmacology; D000897:Anti-Ulcer Agents / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000378:metabolism; D020869:Gene Expression Profiling; D006801:Humans; D006827:Hydantoins / Q000494:pharmacology; D051550:I-kappa B Kinase / Q000037:antagonists & inhibitors / Q000378:metabolism; D016285:Iloprost / Q000494:pharmacology; D053582:Interleukin-1alpha / Q000494:pharmacology*; D048031:JNK Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D008168:Lung / Q000166:cytology / Q000378:metabolism; D020930:MAP Kinase Kinase Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D016595:Misoprostol / Q000494:pharmacology; D020411:Oligonucleotide Array Sequence Analysis; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology; D011459:Prostaglandins E, Synthetic / Q000494:pharmacology; D043562:Receptors, G-Protein-Coupled / Q000819:agonists*; D011982:Receptors, Prostaglandin / Q000819:agonists; D015398:Signal Transduction / Q000187:drug effects*; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism
D000897:Anti-Ulcer Agents; D004791:Enzyme Inhibitors; D006827:Hydantoins; D053582:Interleukin-1alpha; D010975:Platelet Aggregation Inhibitors; D011459:Prostaglandins E, Synthetic; D043562:Receptors, G-Protein-Coupled; D011982:Receptors, Prostaglandin; D016595:Misoprostol; C030669:BW 245C; D000242:Cyclic AMP; D051550:I-kappa B Kinase; D017868:Cyclic AMP-Dependent Protein Kinases; D048031:JNK Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; D020930:MAP Kinase Kinase Kinases; D000527:Alprostadil; C048491:butaprost; D016285:Iloprost; D002118:Calcium