The a9?1 integrin is highly expressed in airway smooth muscle. Mice we have generated lacking this integrin only in smooth muscle cells have marked in vivo airway hyperresponsiveness and lung slices from these mice have increased airway narrowing. We have previously shown that the a9 subunit cytoplasmic domain directly binds the enzyme spermine/spermidine acetyltransferase (SSAT), the rate limiting step in catabolism of higher order polyamines, and that this association and polyamine catabolism are important modulators of a9?1 function. Pharmacologic stabilization of SSAT also augments a9?1-dependent prevention of airway smooth muscle contraction. In the current application we will systematically evaluate the effects of a9?1 on responses of airway smooth muscle to multiple contractile agonists and to isoproterenol-induced relaxation using 4 parallel experimental systems (in vivo AHR, tracheal ring contraction, airway narrowing in lung slices and shortening of airway smooth muscle cells). We will utilize a variety of mutant and chimeric constructs of a9 and SSAT, in vivo and in vitro studies with SSAT knockout mice, catalytically inactive SSAT mutants and genetic and pharmacologic inhibitors to thoroughly examine what contribution interaction of a9 with SSAT makes to this response. Force generation in smooth muscle depends on calcium-dependent actin-myosin cross-bridging and parallel actin polymerization, and we will systematically evaluate the effects of a9?1 on each of these pathways. Because the two major cytosolic effects of higher order polyamines are prevention of potassium efflux through Kir channels and activation of the lipid kinase, PIP5K1?, we will pay special attention to the roles of Kir channels and PIP5K1? in this process. The proposed studies will test the overall hypothesis that ligated a9?1 normally serves as a brake on airway narrowing by concentrating SSAT, catabolizing polyamines and thus inhibiting potassium efflux and/or PIP2 production, resulting in reduced calcium oscillations, decreased actin-myosin cross-bridging and/or impaired actin polymerization. Abnormalities in this pathway, either acquired or genetic, could contribute to diseases such as asthma that are characterized by enhanced airway narrowing.
Project Exaggerated airway narrowing is a central feature of asthma. We have found that mice lacking a specific protein (an integrin) only in smooth muscle have exaggerated airway narrowing. In this proposal we will determine how this integrin normally prevents airway narrowing, a process that could be perturbed in diseases such as asthma.
| Sundaram, Aparna; Chen, Chun; Khalifeh-Soltani, Amin et al. (2017) Targeting integrin ?5?1 ameliorates severe airway hyperresponsiveness in experimental asthma. J Clin Invest 127:365-374 |
| Travis, Mark A; Sheppard, Dean (2014) TGF-? activation and function in immunity. Annu Rev Immunol 32:51-82 |
| Bhattacharya, Mallar; Sundaram, Aparna; Kudo, Makoto et al. (2014) IQGAP1-dependent scaffold suppresses RhoA and inhibits airway smooth muscle contraction. J Clin Invest 124:4895-8 |
| Erle, David J; Sheppard, Dean (2014) The cell biology of asthma. J Cell Biol 205:621-31 |
| Henderson, Neil C; Sheppard, Dean (2013) Integrin-mediated regulation of TGF? in fibrosis. Biochim Biophys Acta 1832:891-6 |
| Friedman, Scott L; Sheppard, Dean; Duffield, Jeremy S et al. (2013) Therapy for fibrotic diseases: nearing the starting line. Sci Transl Med 5:167sr1 |
| Sheppard, Dean (2013) ROCKing pulmonary fibrosis. J Clin Invest 123:1005-6 |
| Henderson, Neil C; Arnold, Thomas D; Katamura, Yoshio et al. (2013) Targeting of ?v integrin identifies a core molecular pathway that regulates fibrosis in several organs. Nat Med 19:1617-24 |
| Sugimoto, Kotaro; Kudo, Makoto; Sundaram, Aparna et al. (2012) The ?v?6 integrin modulates airway hyperresponsiveness in mice by regulating intraepithelial mast cells. J Clin Invest 122:748-58 |
| Chen, Chun; Kudo, Makoto; Rutaganira, Florentine et al. (2012) Integrin ?9?1 in airway smooth muscle suppresses exaggerated airway narrowing. J Clin Invest 122:2916-27 |
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