The mechanical stimulation of airway smooth muscle (ASM) that occurs during stretch and oscillation of the airways during breathing is critical for maintaining a normal low level of airway reactivity. Furthermore, chronic loading of the airways by continuous positive end expiratory pressure (CPAP) in vivo or prolonged loading of ASM in vitro reduces airway responsiveness and alters ASM phenotype, suggesting that CPAP may act as a potential therapy for asthma. Pathophysiologic conditions also induce alterations in mechanical forces imposed on the airways during breathing and in airway tissue structure and extracellular matrix composition. These environmental changes modulate ASM phenotype and its responses to physiologic stimuli, but the mechanisms for the effects of mechanical forces and the extracellular environment on the properties of ASM are not understood.The adhesion junctions that connect cells within tissues to the extracellular matrix are composed of large multiprotein complexes termed "adhesomes". Adhesomes provide mechanical coupling between cells and their matrix environment, and they also enable cells to sense and respond to changes in their surrounding milieu. The contractile stimulation of ASM triggers the recruitment of structural and signaling proteins to adhesome complexes, and adhesome signaling complex components are critical for the normal physiologic responses of ASM to stimulation. However, the molecular mechanisms that catalyze adhesome assembly in response to ASM stimulation and regulate the organization and activation of component proteins are not known. The proposed studies will explore a novel molecular hypothesis for the assembly and activation of adhesome proteins in ASM in response to contractile and inflammatory stimuli. These processes are fundamental components of the signal transduction process in ASM, and provide a mechanism for ASM to integrate its physiologic responses to humoral and environmental stimuli. The proposed studies will employ ASM tissues and freshly dissociated differentiated ASM cells to address three Specific Aims: 1) Determine the roles of RhoA and NM myosin II in regulating the assembly and activation of adhesome signaling complexes, cytoskeletal dynamics and ASM contractility. 2) Determine how contractile stimulation regulates the activation of integrin adhesome complex proteins. 3) Determine the roles of RhoA, NM myosin II and adhesome protein activation in regulating the responses of ASM to inflammatory and humoral stimuli and their modulation by mechanical forces and extracellular matrix proteins. These studies will provide new insights into the molecular mechanisms of signal transduction in ASM that are likely to be broadly relevant to other cells and tissues, and are important for understanding how mechanical forces interact with drugs and hormones to regulate the physiologic responses in ASM tissues. They will provide a basis for designing new therapeutic approaches involving mechanical modalities as well as pharmacologic therapy for the treatment of airways disease.

Public Health Relevance

The physiologic responses of airway smooth muscle to endogenous hormones and exogenous stimuli, including therapeutic agent, are affected by the external forces imposed on the airway tissues during breathing and the local environment of the cells. The project will describe new molecular processes by which airway smooth muscle integrates mechanical and local environmental stimuli with humoral stimuli to determine its physiologic responses and properties. These studies are important for understanding how mechanical forces interact with drugs and hormones to regulate the physiologic responses in airway smooth muscle tissues, and will provide a basis for designing new therapeutic approaches involving mechanical modalities as well as pharmacologic therapy for the treatment of airways disease.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL029289-27
Application #
8606756
Study Section
Lung Cellular, Molecular, and Immunobiology Study Section (LCMI)
Program Officer
Banks-Schlegel, Susan P
Project Start
1989-07-01
Project End
2017-01-31
Budget Start
2014-02-01
Budget End
2015-01-31
Support Year
27
Fiscal Year
2014
Total Cost
$351,000
Indirect Cost
$126,000
Name
Indiana University-Purdue University at Indianapolis
Department
Physiology
Type
Schools of Medicine
DUNS #
603007902
City
Indianapolis
State
IN
Country
United States
Zip Code
46202
Wu, Yidi; Huang, Youliang; Gunst, Susan J (2016) Focal adhesion kinase (FAK) and mechanical stimulation negatively regulate the transition of airway smooth muscle tissues to a synthetic phenotype. Am J Physiol Lung Cell Mol Physiol 311:L893-L902
Zhang, Wenwu; Huang, Youliang; Gunst, Susan J (2016) p21-Activated kinase (Pak) regulates airway smooth muscle contraction by regulating paxillin complexes that mediate actin polymerization. J Physiol 594:4879-900
Wu, Yidi; Gunst, Susan J (2015) Vasodilator-stimulated phosphoprotein (VASP) regulates actin polymerization and contraction in airway smooth muscle by a vinculin-dependent mechanism. J Biol Chem 290:11403-16
Zhang, Wenwu; Huang, Youliang; Wu, Yidi et al. (2015) A novel role for RhoA GTPase in the regulation of airway smooth muscle contraction. Can J Physiol Pharmacol 93:129-36
Huang, Youliang; Day, Richard N; Gunst, Susan J (2014) Vinculin phosphorylation at Tyr1065 regulates vinculin conformation and tension development in airway smooth muscle tissues. J Biol Chem 289:3677-88
Lott, Jeremy M; Sehra, Sarita; Mehrotra, Purvi et al. (2013) Type V collagen-induced tolerance prevents airway hyperresponsiveness. Am J Respir Crit Care Med 187:454-7
Xue, Z; Zhang, W; Desai, L P et al. (2013) Increased mechanical strain imposed on murine lungs during ventilation in vivo depresses airway responsiveness and activation of protein kinase Akt. J Appl Physiol (1985) 114:1506-10
Busk, Michael; Busk, Nancy; Puntenney, Paula et al. (2013) Use of continuous positive airway pressure reduces airway reactivity in adults with asthma. Eur Respir J 41:317-22
Touw, Ketrija; Chakraborty, Saikat; Zhang, Wenwu et al. (2012) Altered calcium signaling in colonic smooth muscle of type 1 diabetic mice. Am J Physiol Gastrointest Liver Physiol 302:G66-76
Hoover, Wyn C; Zhang, Wenwu; Xue, Zhidong et al. (2012) Inhibition of p21 activated kinase (PAK) reduces airway responsiveness in vivo and in vitro in murine and human airways. PLoS One 7:e42601

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