Transcriptional response to persistent beta2-adrenergic receptor signaling reveals regulation of phospholamban, which alters airway contractility.
Beta(2)-Adrenergic receptors (beta(2)AR) are expressed on airway smooth muscle cells and act to relax the airway on activation by beta-agonists. These agents are utilized for treating asthma but are associated with adverse outcomes. To ascertain the effects of persistent beta(2)AR activation on gene expression, cultured airway smooth muscle cells derived from wild-type (WT) and transgenic mice overexpressing beta(2)AR were subjected to DNA microarray analysis; 319 genes were increased and 164 were decreased. Differential expression was observed in genes from 22 Gene Ontology Slim categories, including those associated with ion transport and calcium ion binding. A 60% decrease (P = 0.008) in phospholamban (PLN), an intracellular Ca(2+) concentration ([Ca(2+)]i)-handling protein that is at a signaling nodal point in cardiomyocytes, was observed in beta(2)AR-overexpressing cells and confirmed at the protein level. To isolate the physiological effect of decreased PLN in airway smooth muscle, airway contraction and relaxation responses were studied in WT and PLN(-/-) mice. PLN(-/-) mice had a markedly reduced constrictive response to methacholine. In contrast, the bronchodilatory effect of beta-agonist was not different between WT and PLN(-/-) mice. These results revealed an unanticipated therapeutic effect of beta-agonists, PLN downregulation, which acts to decrease airway hyperreactivity. Thus agents that inhibit PLN may act synergistically with the bronchodilating action of beta-agonists. A number of other genes related to [Ca(2+)]i are also differentially regulated by beta(2)AR activity, some of which may act to oppose, or augment, the efficacy of chronic beta-agonists. These genes or pathways may also represent additional targets in the treatment of asthma and related obstructive lung diseases.
McGraw
Dennis W
DW
Pulmonary and Critical Care Medicine, University of Cincinnati College of Medicine, Cincinnati, OH, USA.
Fogel
Kevin M
KM
Kong
Sue
S
Litonjua
Augusto A
AA
Kranias
Evangelia G
EG
Aronow
Bruce J
BJ
Liggett
Stephen B
SB
D000403:Airway Resistance / Q000187:drug effects; D000818:Animals; D018927:Anti-Asthmatic Agents / Q000494:pharmacology; D016084:Bronchoconstriction / Q000187:drug effects / Q000235:genetics*; D016085:Bronchoconstrictor Agents / Q000494:pharmacology; D001993:Bronchodilator Agents / Q000494:pharmacology; D020013:Calcium Signaling; D002135:Calcium-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D002478:Cells, Cultured; D020869:Gene Expression Profiling; D007545:Isoproterenol / Q000494:pharmacology; D016210:Methacholine Chloride / Q000494:pharmacology; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D009130:Muscle, Smooth / Q000187:drug effects* / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D018343:Receptors, Adrenergic, beta-2 / Q000235:genetics / Q000502:physiology*; D015398:Signal Transduction / Q000187:drug effects; D014132:Trachea / Q000166:cytology; D014158:Transcription, Genetic / Q000187:drug effects; D000099211:Phospholamban
D018927:Anti-Asthmatic Agents; D016085:Bronchoconstrictor Agents; D001993:Bronchodilator Agents; D002135:Calcium-Binding Proteins; D007545:Isoproterenol; D016210:Methacholine Chloride; D018343:Receptors, Adrenergic, beta-2; D000099211:Phospholamban