- PROJECT 2 During the previous award cycle, we identified mechanisms by which primary airway epithelial cells (AECs) synthesize heavy chain (HC) modified hyaluronan (HA) apical rafts, and airway smooth muscle cells (ASMCs) synthesize an extracellular matrix of HA cables, which are leukocyte-adhesive or what we have also termed as pathologic-HA. Our studies in the Program revealed the importance of this pathologic-HA matrix in the genesis of inflammation, airway remodeling and hyperresponsiveness of asthma. Within the airway, there is constant interaction between the closely coupled ASMCs and overlying epithelium, and increasing data indicate a primary role for epithelium and smooth muscle in the origins of inflammation through production of early mediators. In this continuation of Project 2, we propose to investigate if airway epithelial production of NO and cytokines promotes the accumulation of a pathologic-HA matrix by the smooth muscle in asthma. In preliminary data, we show that the airway epithelium responds to prototypic Th2 cytokines or cytokine mixes with the production of HA and induction of iNOS. Utilizing our novel co-culture system, we show that the ASMCs can respond to epithelial derived NO with the production of leukocyte-adhesive HA cables. We also show that the major contributor to HA production is via the hyaluronan synthase 2 (HAS2). Collaborative studies of NO regulation and signaling mechanisms, which are investigated by other members of this Program, support our experiments to define mechanisms of pathological HA formation, including NO metabolism (Project 1), specific protein thiol nitrosylation (Project 3), and NO activation pathways (Project 4). Thus, in this continuation, we propose to test the hypothesis that NO and HCs produced by the asthmatic airway epithelium promote formation of a pathologic, leukocyte-adhesive HC-HA matrix by ASMCs, through mechanisms that appear to be sGC-independent NO recognition pathways. To test this hypothesis, we plan to identify pathways leading to airway epithelial cell production of HA [Aim 1]. We also plan to identify pathways of ASMC production of a leukocyte-adhesive HA matrix [Aim 2]. Finally, we will determine the contribution of NO and HA on the development of inflammation, AHR, and remodeling in vivo [Aim 3]. Project 2 depends on the collaborative interactions and complementary skills, expertise, and resources of the Program to test hypotheses. Program Cores support novel murine models, technically challenging assays, and shared clinical samples in order to facilitate translational and mechanistic experiments in Project 2 not otherwise possible.

Public Health Relevance

- PROJECT 2 Asthma is a chronic inflammation of the airways in which the local resident cells of the airway play a dominant role in the initiation and perpetuation of the inflammatory response. The chronic inflammation can lead to progressive airway changes over time that may interfere with routine activities of daily living. Our project will determine the effects of local mediators on the development of asthma and identify possible targets for future therapeutic interventions.

National Institute of Health (NIH)
National Heart, Lung, and Blood Institute (NHLBI)
Research Program Projects (P01)
Project #
Application #
Study Section
Heart, Lung, and Blood Initial Review Group (HLBP)
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Cleveland Clinic Lerner
United States
Zip Code
Johnson, Collin G; Stober, Vandy P; Cyphert-Daly, Jaime M et al. (2018) High molecular weight hyaluronan ameliorates allergic inflammation and airway hyperresponsiveness in the mouse. Am J Physiol Lung Cell Mol Physiol :
Majors, Alana K; Chakravarti, Ritu; Ruple, Lisa M et al. (2018) Nitric oxide alters hyaluronan deposition by airway smooth muscle cells. PLoS One 13:e0200074
Sweeny, Elizabeth A; Singh, Anuradha Bharara; Chakravarti, Ritu et al. (2018) Glyceraldehyde-3-phosphate dehydrogenase is a chaperone that allocates labile heme in cells. J Biol Chem 293:14557-14568
Reichard, Andrew; Wanner, Nicholas; Stuehr, Eric et al. (2018) Quantification of airway fibrosis in asthma by flow cytometry. Cytometry A 93:952-958
Asosingh, Kewal; Weiss, Kelly; Queisser, Kimberly et al. (2018) Endothelial cells in the innate response to allergens and initiation of atopic asthma. J Clin Invest 128:3116-3128
Allawzi, Ayed M; Vang, Alexander; Clements, Richard T et al. (2018) Activation of Anoctamin-1 Limits Pulmonary Endothelial Cell Proliferation via p38-Mitogen-activated Protein Kinase-Dependent Apoptosis. Am J Respir Cell Mol Biol 58:658-667
Reichard, Andrew; Asosingh, Kewal (2018) The role of mitochondria in angiogenesis. Mol Biol Rep :
Ghosh, Arnab; Garee, Greer; Sweeny, Elizabeth A et al. (2018) Hsp90 chaperones hemoglobin maturation in erythroid and nonerythroid cells. Proc Natl Acad Sci U S A 115:E1117-E1126
Comhair, Suzy A A; Bochenek, Grazyna; Baicker-McKee, Sara et al. (2018) The utility of biomarkers in diagnosis of aspirin exacerbated respiratory disease. Respir Res 19:210
Stober, Vandy P; Johnson, Collin G; Majors, Alana et al. (2017) TNF-stimulated gene 6 promotes formation of hyaluronan-inter-?-inhibitor heavy chain complexes necessary for ozone-induced airway hyperresponsiveness. J Biol Chem 292:20845-20858

Showing the most recent 10 out of 123 publications