The development of subepithelial fibrosis represents an important feature of airway remodeling in asthma, and a critical role for epithelial cells herei is emerging. The pro-fibrotic cytokine, transforming growth factor beta-1 (TGF-?1) plays a cardinal role in fibrogenesis. The outcome of TGF-?1 signaling is highly dependent upon cooperation with other signaling pathways. We have recently identified a critical role for c-Jun-N-terminal- kinase 1 (JNK1) in augmenting the pro-fibrotic effects of TGF-?1, in association with the causation of a mesenchymal transition of airway epithelial cells (EMT). Specifically, we have demonstrated that: 1) JNK is predominantly activated in airway epithelium from ovalbumin-sensitized and challenged mice;2) mice lacking JNK1 (JNK1-/-) fail to induce mesenchymal genes or develop fibrosis in response to ovalbumin, TGF-?1, or bleomycin;3) TGF-?1-induced EMT requires JNK1, through JNK1-dependent phosphorylation of SMAD3 in the linker domain. These data, suggest a critical role of airway epithelium-derived JNK1-dependent signals in orchestrating airways fibrosis. The hypothesis addressed herein is that activation of JNK1 in the airway epithelium is required for the development of subepithelial fibrosis in house dust mite- induced airways disease by promoting epithelial-mesenchymal transition (EMT). Specifically, we hypothesize that JNK1 enhances TGF-?1 signaling via phosphorylation of SMAD3 in the linker domain, which enhances the competency of SMAD3 to decrease expression of let-7 microRNA (miRNA). Decreases in let-7 miRNA in turn enhances expression of the high mobility group A2 (HMGA2) gene, a proximal regulator of EMT, events that lead to subepithelial collagen deposition.
In Specific Aim 1 we will determine the importance of JNK1-dependent phosphorylation of SMAD3 in the linker domain in repressing let-7 microRNA (miRNA) in lung epithelial cells, and subsequent enhancement of the high mobility group A2 (HMGA2) gene. We will also analyze expression of let-7 miRNA and HMGA2 in bronchial epithelial cells derived from asthmatics, compared to controls, and determine the susceptibility of asthmatic and normal epithelial cells to TGF-?1-induced EMT, and elucidate the role of JNK and SMAD3 therein.
In Specific Aim 2 we will elucidate whether activation of JNK1 within the airway epithelium is critical in the orchestration of house dust mite-induced epithelial to mesenchymal transition, airways fibrosis, and altered respiratory mechanics in vivo, and utilize mice that globally lack JNK1, or specifically within the bronchiolar epithelium, following and CRE-recombinase mediated ablation.
In Specific Aim 3 we will determine the importance of SMAD3 linker domain phosphorylation and resultant decreases of let-7g miRNA in promoting epithelial to mesenchymal transition, airways fibrosis, and altered respiratory mechanics in vivo, via the creation of TetOP-FLAG-SMAD3-EPSM mice, which are refractory to phosphorylation by JNK in the linker domain, and TetOP-FLAG-SMAD3-wild type (WT) transgenic mice, as a control. We will assess the impact of delivery of let-7g pre-miRNA on HDM-induced remodeling.
This project will demonstrate the importance of airway epithelial cells which line the airways for the development of a scar in the lung. We have demonstrated that in mice which lack a specific signaling molecule, known as c-Jun-N-terminal kinase, are resistant to the development of a lung scar. In this project we will use sophisticated genetic tools to specifically remove this signaling protein only from epithelial cells, but not othr cell types in the lung, so that we can confirm the importance of c-jun-N-terminal kinase in epithelial cells only, for development of a scar. We will also determine the mechanism whereby c-jun-N-terminal kinase causes its scarring effects. We will also determine whether of c-jun-N-terminal kinase is active in airway epithelial cells isolated from patients with asthma, compared to control subjects. This project is important in that it will demonstrate that therapeutics to prevent scars should be directed to epithelial cells, and may provide new opportunities to stop the process of scarring in the future.
|Hoffman, Sidra M; Chapman, David G; Lahue, Karolyn G et al. (2016) Protein disulfide isomerase-endoplasmic reticulum resident protein 57 regulates allergen-induced airways inflammation, fibrosis, and hyperresponsiveness. J Allergy Clin Immunol 137:822-832.e7|
|Hoffman, Sidra M; Qian, Xi; Nolin, James D et al. (2016) Ablation of Glutaredoxin-1 Modulates House Dust Mite-Induced Allergic Airways Disease in Mice. Am J Respir Cell Mol Biol 55:377-86|
|Jones, Jane T; Qian, Xi; van der Velden, Jos L J et al. (2016) Glutathione S-transferase pi modulates NF-ÎºB activation and pro-inflammatory responses in lung epithelial cells. Redox Biol 8:375-82|
|Hristova, Milena; Habibovic, Aida; Veith, Carmen et al. (2016) Airway epithelial dual oxidase 1 mediates allergen-induced IL-33 secretion and activation of type 2 immune responses. J Allergy Clin Immunol 137:1545-1556.e11|
|Hoffman, Sidra; Nolin, James; McMillan, David et al. (2015) Thiol redox chemistry: role of protein cysteine oxidation and altered redox homeostasis in allergic inflammation and asthma. J Cell Biochem 116:884-92|
|Li, Shannon; Aliyeva, Minara; Daphtary, Nirav et al. (2014) Antigen-induced mast cell expansion and bronchoconstriction in a mouse model of asthma. Am J Physiol Lung Cell Mol Physiol 306:L196-206|
|Chapman, David G; Tully, Jane E; Nolin, James D et al. (2014) Animal models of allergic airways disease: where are we and where to next? J Cell Biochem 115:2055-64|
|van der Velden, Jos L J; Hoffman, Sidra M; Alcorn, John F et al. (2014) Absence of c-Jun NH2-terminal kinase 1 protects against house dust mite-induced pulmonary remodeling but not airway hyperresponsiveness and inflammation. Am J Physiol Lung Cell Mol Physiol 306:L866-75|
|Janssen-Heininger, Yvonne M W; Nolin, James D; Hoffman, Sidra M et al. (2013) Emerging mechanisms of glutathione-dependent chemistry in biology and disease. J Cell Biochem 114:1962-8|
|van der Velden, Jos L J; Guala, Amy S; Leggett, Susan E et al. (2012) Induction of a mesenchymal expression program in lung epithelial cells by wingless protein (Wnt)/Ã½Ã½-catenin requires the presence of c-Jun N-terminal kinase-1 (JNK1). Am J Respir Cell Mol Biol 47:306-14|
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