Our objective is to define cellular and molecular mechanisms by which airway smooth muscle cells contribute to airway hyperreactivity. Initial studies focused on acute, postjunctional events that increase smooth muscle hyperresponsiveness. New studies are proposed to define the roles of ERK and p38/RK mitogen-activated protein kinases (MAP kinases) in canine tracheal smooth muscle contraction and airway wall remodeling. The hypothesis is that MAP kinases are components of protein kinase cascades controlling contractile protein function, contractile protein gene expression and structure of the cytoskeleton. Coupling of ERK MAP kinases to contractile system function will be studied by: i) Assaying agonist and Ca2+-dependence of ERK MAP kinase activation, ii) Determining the functional consequences of ERK activation, and iii) Sequencing ERK isoforms expressed in differentiated canine tracheal smooth muscle. Coupling of ERK MAP kinases to contractile protein gene expression in cultured tracheal myocytes will be investigated by: i) Determining whether chronic receptor activation regulates both ERK MAP kinase and contractile protein expression, and iii) Test the necessity of MAP kinases in maintaining the contractile phenotype by inhibiting MAP kinase expression with antisense oligodeoxynucleotides. Coupling of p38/RK MAP kinase to cytoskeletal structure may occur through activation of MAP kinase-activated kinase-II (MAPKAP II) which phosphorylates the 27 kDa heat shock protein (HSP27). To test this hypothesis we will: i) Assay tyrosine phosphorylation of p38/RK MAP kinase, activation of MAPKAP II and phosphorylation of HSP27, ii) Establish actin binding activity of HSP27, iii) Determine whether treatments that increase HSP27 phosphorylation also increase actin polymerization, and iv) Investigate the function of HSP27 in migration and shortening of cultured myocytes. We propose to define the roles of ERK and p38/RK MAP kinases in contractile and proliferative phenotypes of airway smooth muscle, which may be valuable in designing strategies to inhibit or reverse smooth muscle hyperreactivity and airway wall remodeling that occurs in asthma.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL048183-08
Application #
6183720
Study Section
Lung Biology and Pathology Study Section (LBPA)
Project Start
1992-04-01
Project End
2001-03-31
Budget Start
2000-04-01
Budget End
2001-03-31
Support Year
8
Fiscal Year
2000
Total Cost
$200,148
Indirect Cost
Name
University of Nevada Reno
Department
Pharmacology
Type
Schools of Medicine
DUNS #
146515460
City
Reno
State
NV
Country
United States
Zip Code
89557
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Gerthoffer, William T (2005) Signal-transduction pathways that regulate visceral smooth muscle function. III. Coupling of muscarinic receptors to signaling kinases and effector proteins in gastrointestinal smooth muscles. Am J Physiol Gastrointest Liver Physiol 288:G849-53
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Singer, Cherie A; Baker, Kimberly J; McCaffrey, Alan et al. (2003) p38 MAPK and NF-kappaB mediate COX-2 expression in human airway myocytes. Am J Physiol Lung Cell Mol Physiol 285:L1087-98
Yamboliev, I A; Chen, J; Gerthoffer, W T (2001) PI 3-kinases and Src kinases regulate spreading and migration of cultured VSMCs. Am J Physiol Cell Physiol 281:C709-18
Gerthoffer, W T; Gunst, S J (2001) Invited review: focal adhesion and small heat shock proteins in the regulation of actin remodeling and contractility in smooth muscle. J Appl Physiol 91:963-72
Hedges, J C; Singer, C A; Gerthoffer, W T (2000) Mitogen-activated protein kinases regulate cytokine gene expression in human airway myocytes. Am J Respir Cell Mol Biol 23:86-94
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Hedges, J C; Dechert, M A; Yamboliev, I A et al. (1999) A role for p38(MAPK)/HSP27 pathway in smooth muscle cell migration. J Biol Chem 274:24211-9
Hedges, J C; Yamboliev, I A; Ngo, M et al. (1998) p38 mitogen-activated protein kinase expression and activation in smooth muscle. Am J Physiol 275:C527-34

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