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Joo-heon Yoon - One of the best experts on this subject based on the ideXlab platform.
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proprotein convertase 5 6a is associated with bone morphogenetic protein 2 induced squamous Cell differentiation
Clinical and Translational Allergy, 2015Co-Authors: Joo-heon Yoon, Sangnam LeeAbstract:Results Using a retinoic acid deficiency-induced squamous metaplasia model of HNECs, we observed a significant increase in the expression of PC5/6A, a PC member, and BMP-2, a candidate substrate for PC5/6A. Specific lentiviral shRNA-mediated PC5/6A knockdown decreased BMP-2 expression and maturation, decreased expression of squamous Cell markers, and increased expression of Ciliated Cell markers. Dec-RVKR-CMK, a PC inhibitor, and LDN-193189, a BMP receptor inhibitor, suppressed squamous differentiation, promoted mucociliary differentiation, and down-regulated the BMP-2/Smad1/5/8/p38 signalling pathways. Dec-RVKRCMK also decreased expression of PC5/6A, but not furin, another PC member, suggesting the involvement of PC5/6A in squamous differentiation of HNECs. Overexpression of PC5/6A and BMP-2 in the human nasal epithelial Cell line RPMI-2650 demonstrated that PC5/6A can activate BMP-2. Under retinoic acid-sufficient culture conditions for mucociliary differentiation of HNECs, short-term expression of PC5/6A by the adenovirus system and addition of exogenous BMP-2 induced squamous differentiation. Furthermore, PC5/6A and BMP-2 were highly expressed in metaplastic squamous epithelium of human nasal polyps.
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proprotein convertase 5 6a is associated with bone morphogenetic protein 2 induced squamous Cell differentiation
American Journal of Respiratory Cell and Molecular Biology, 2014Co-Authors: Sangnam Lee, Dahyung Lee, Min Goo Lee, Joo-heon YoonAbstract:Squamous metaplasia in airway epithelium is a pathological process arising from abnormal remodeling/repair responses to injury. Proteolytic maturation of many growth and differentiation factors involved in tissue remodeling is controlled by proprotein convertases (PCs). However, the role of these convertases in airway remodeling remains poorly understood. Using a retinoic acid deficiency–induced squamous metaplasia model of cultured human nasal epithelial Cells (HNECs), we observed a significant increase in the expression of PC5/6A, a PC member, and bone morphogenetic protein-2 (BMP-2), a candidate substrate for PC5/6A. Specific lentiviral short hairpin RNA–mediated PC5/6A knockdown decreased BMP-2 expression and maturation, decreased expression of squamous Cell markers, and increased expression of Ciliated Cell markers. Decanoyl-Arg-Val-Lys-Arg-chloromethylketone (Dec-RVKR-CMK), a PC inhibitor, and LDN-193189, a BMP receptor inhibitor, suppressed squamous differentiation, promoted mucociliary differentia...
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MUC8 as a Ciliated Cell Marker in Human Nasal Epithelium
Acta oto-laryngologica, 2005Co-Authors: Chang Hoon Kim, Hyun Jik Kim, Kyoung Seob Song, Je Kyung Seong, Kyung-su Kim, Jeung Gweon Lee, Joo-heon YoonAbstract:Conclusions This study indicates that MUC8 protein is expressed in Ciliated Cells from human nasal epithelial Cells and is upregulated by IL-1β treatment. These results suggest that MUC8 gene and protein expression levels could be used as a Ciliated Cell marker in human nasal epithelium. Objectives To examine MUC8 mRNA expression patterns according to the mucociliary differentiation of normal human nasal epithelial (NHNE) Cells, and to investigate the localization of MUC8 proteins in nasal polyps. Material and methods Passage-2 NHNE Cells were cultured using an air–liquid interface technique. On Days 2, 7, 14 and 28 after confluence, Ciliated Cells were counted by means of cytospin slide immunostaining using H6C5 and β-tubulin, and MUC8 mRNA levels were determined using real-time quantitative polymerase chain reaction (PCR). After synthesizing polyclonal anti-MUC8 peptide antibodies, MUC8 immunostaining was performed using nasal polyps. MUC8 mRNA and protein levels were determined in NHNE Cells treated wi...
S T Holgate - One of the best experts on this subject based on the ideXlab platform.
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Characterization of Ciliated Bronchial Epithelium 1, a Ciliated Cell–Associated Gene Induced During Mucociliary Differentiation
2013Co-Authors: Hajime Yoshisue, Sarah M Puddicombe, Susan J Wilson, Hans Michael Haitchi, Robert M Powell, David I Wilson, Donna E Davies, Anita P, Ann E. Berger, S T HolgateAbstract:Lung epithelial structure is altered in asthma; however, the precise mechanisms underlying epithelial repair, including differentiation from basal to columnar epithelial Cells, are not well defined. In the course of random sequencing of a cDNA library from human lung biopsies, we have identified a novel gene, Ciliated bronchial epithelium 1 (CBE1). Expression of CBE1 was induced during in vitro differentiation of bronchial epithelial Cells. Synchronous expression with tektin and hepatocyte nuclear factor 3/forkhead homologue 4, downregulation by interleukin-13, and its tissue distribution strongly suggested that CBE1 is associated with Ciliated Cells. Two isoforms of the 0.7-kb full-length cDNA were identified, resulting in open reading frames with different carboxyl termini, with no homology to known proteins. Expression of CBE1 in Ciliated epithelial Cells was confirmed by immunohistochemistry. Quantitative reverse transcription–polymerase chain reaction analysis using bronchia
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characterization of Ciliated bronchial epithelium 1 a Ciliated Cell associated gene induced during mucociliary differentiation
American Journal of Respiratory Cell and Molecular Biology, 2004Co-Authors: Hajime Yoshisue, Sarah M Puddicombe, Susan J Wilson, Hans Michael Haitchi, Robert M Powell, David I Wilson, Anita Pandit, Ann Berger, Donna E Davies, S T HolgateAbstract:Lung epithelial structure is altered in asthma; however, the precise mechanisms underlying epithelial repair, including differentiation from basal to columnar epithelial Cells, are not well defined. In the course of random sequencing of a cDNA library from human lung biopsies, we have identified a novel gene, Ciliated bronchial epithelium 1 (CBE1). Expression of CBE1 was induced during in vitro differentiation of bronchial epithelial Cells. Synchronous expression with tektin and hepatocyte nuclear factor 3/forkhead homologue 4, downregulation by interleukin-13, and its tissue distribution strongly suggested that CBE1 is associated with Ciliated Cells. Two isoforms of the 0.7-kb full-length cDNA were identified, resulting in open reading frames with different carboxyl termini, with no homology to known proteins. Expression of CBE1 in Ciliated epithelial Cells was confirmed by immunohistochemistry. Quantitative reverse transcription–polymerase chain reaction analysis using bronchial biopsies showed no difference of expression of CBE1 between normal subjects and subjects with asthma. Expression studies showed that CBE1 is nuclear- or perinuclear-localized, depending on Cell type. Regulated expression during differentiation and the subCellular localization of CBE1 suggest that it may play an important role in the differentiation and/or function of Ciliated Cells in human airways. Mammalian lung epithelium is composed of a variety of Cell populations from proximal to distal airways. The individual characteristics of the subtypes of the Cells that make up the airway epithelium not only create an effective physical barrier against various noxious substances, but also a highly sophisticated host defense system by producing and releasing a large number of chemical mediators and cytokines (reviewed in Refs. 1 and 2). The epithelium of the upper airways is a continuous layer consisting mainly of three Cell types: basal, goblet, and Ciliated epithelial Cells, the latter two of which make up a suprabasal columnar structure and are necessary for mucociliary clearance. Goblet Cells synthesize mucus, providing the surface of the epithelium with a protective cover, and Ciliated Cells are responsible for propelling the mucus secretions toward the pharynx.
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Ciliated Cell damage in the bronchial epithelium of asthmatics and non asthmatics
Clinical & Experimental Allergy, 1993Co-Authors: Stephen Montefort, Ratko Djukanovic, S T Holgate, W R RocheAbstract:The importance of bronchial epithelial shedding in the pathogenesis of asthma has been highlighted by many investigators as a potential mechanism for bronchial hyperresponsiveness. It has been suggested that this disruption is the result of cytotoxic injury leading to shedding of damaged Cells. To investigate whether damaged Ciliated epithelial Cells can be detected within the bronchial mucosa, we used tannic acid which only permeates disrupted Cellular membranes, as a marker of Cell damage. Bronchial biopsies from seven asthmatic and six normal subjects, were processed in tannic acid prior to preparation and sectioning for electronmicroscopic examination. Ciliated epithelial Cells staining darkly with tannic acid were seen to comprise a similar proportion of the intact portion of bronchial epithelium in both normals and asthmatics (medians 31% vs 40%). We suggest that Ciliated epithelial Cells are not shed from the bronchial mucosa immediately after damage and that mechanisms other than granulocyte-mediated cytotoxicity may account for epithelial disruption in asthma, possibly involving the selective damage or reduced expression of intraepithelial interCellular adhesion molecules.
Steven L. Brody - One of the best experts on this subject based on the ideXlab platform.
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Correspondence should be addressed to:
2013Co-Authors: Yong Zhang, William T Roswit, Steven L. Brody, Guangming Huang, Laurie P. Shornick, James M. Shipley, Michael J. HoltzmanAbstract:Ciliated airway epithelial Cells are critical for mucosal barrier function, including host defense against pathogens. This Cell population is often the primary target and thereby the first line of defense against many common respiratory viruses. It is also the precursor for mucous Cells and thereby promotes mucociliary clearance of infectious and other noxious agents. Cells with motile cilia in other organs, e.g., brain and reproductive organs, may also have roles in development and reproduction. However, definitive proof of Ciliated Cell function is hampered by the lack of strategies to specifically target this Cell population for loss of function in vivo. To this end, Celltype specific gene promoters have been combined with the Cre/LoxP system to disrupt genes in airway and alveolar epithelial Cell populations expressing surfactant protein C (SP-C) or Clara Cell secretory protein (CCSP). By contrast, an analogous system to disrupt gene function in Ciliated airway epithelial Cells was still needed. Here we report the generation and analysis of mouse lines with a FOXJ1 promoter driving the Cre recombinase and show that this system mediates genomic recombination specifically in Ciliated Cells. The pattern of recombination recapitulates endogenous FOXJ1 promoter function, being restricted to Ciliated Cells present in pulmonary airways as well as choroid plexus, ependyma, oviduct, and testis. This transgeni
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RhoA-mediated apical actin enrichment is required for ciliogenesis and promoted by Foxj1.
Journal of Cell Science, 2007Co-Authors: Jiehong Pan, Tao Huang, Yingjian You, Steven L. BrodyAbstract:Programs that direct Cellular differentiation are dependent on the strict temporal expression of regulatory factors that can be provided by Rho GTPases. Ciliogenesis is a complex sequence of events involving the generation and docking of basal bodies at the apical membrane, followed by ciliary axoneme generation. Although a cilia proteome has been assembled, programs that direct Ciliated Cell differentiation are not well established, particularly in mammalian systems. Using mouse primary culture airway epithelial Cells, we identified a critical stage of ciliogenesis requiring the temporal establishment of an apical web-like structure of actin for basal body docking and subsequent axoneme growth. Apical web formation and basal body docking were prevented by interruption of actin remodeling and were dependent on RhoA activation. Additional evidence for this program was provided by analysis of Foxj1-null mice that failed to dock basal bodies and lacked apical actin. Foxj1 expression coincided with actin web formation, activated RhoA and RhoB, and persisted despite RhoA inhibition, suggesting that Foxj1 promoted RhoA during ciliogenesis. Apical ezrin localization was also dependent on Foxj1, actin remodeling, and RhoA, but was not critical for ciliogenesis. Thus, temporal Foxj1 and RhoA activity are essential regulatory events for cytoskeletal remodeling during mammalian ciliogenesis.
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trefoil factor family 3 peptide promotes human airway epithelial Ciliated Cell differentiation
American Journal of Respiratory Cell and Molecular Biology, 2007Co-Authors: Pierre Lesimple, Steven L. Brody, Isabelle Van Seuningen, Mariepierre Buisine, Mariechristine Copin, Margitta Hinz, Werner Hoffmann, Rodolphe Hajj, Christelle Coraux, Edith PuchelleAbstract:Human airway surface epithelium is frequently damaged by inhaled factors (viruses, bacteria, xenobiotic substances) as well as by inflammatory mediators that contribute to the shedding of surface epithelial Cells. To regain its protective function, the epithelium must rapidly repair and redifferentiate. The Trefoil Factor Family (TFF) peptides are secretory products of many mucous Cells. TFF3, the major TFF in the airways, is able to enhance airway epithelial Cell migration, but the role of this protein in differentiation has not been defined. To identify the specific role of TFF3 in the differentiation of the human airway surface epithelium, we analyzed the temporal expression pattern of TFF3, MUC5AC, and MUC5B mucins (goblet Cells) and Ciliated Cell markers β-tubulin (cilia) and FOXJ1 (ciliogenesis) during human airway epithelial regeneration using in vivo humanized airway xenograft and in vitro air–liquid interface (ALI) culture models. We observed that TFF3, MUC5AC, MUC5B, and Ciliated Cell markers we...
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blocking airway mucous Cell metaplasia by inhibiting egfr antiapoptosis and il 13 transdifferentiation signals
Journal of Clinical Investigation, 2006Co-Authors: Jeffrey W. Tyner, John T Battaile, Mark R Pelletier, Jeffrey D Morton, Melanie S Spoor, William T Roswit, Mario Castro, Alexander G Patterson, Anand C. Patel, Steven L. BrodyAbstract:Epithelial hyperplasia and metaplasia are common features of inflammatory and neoplastic disease, but the basis for the altered epithelial phenotype is often uncertain. Here we show that long-term Ciliated Cell hyperplasia coincides with mucous (goblet) Cell metaplasia after respiratory viral clearance in mouse airways. This chronic switch in epithelial behavior exhibits genetic susceptibility and depends on persistent activation of EGFR signaling to PI3K that prevents apoptosis of Ciliated Cells and on IL-13 signaling that promotes transdifferentiation of Ciliated to goblet Cells. Thus, EGFR blockade (using an irreversible EGFR kinase inhibitor designated EKB-569) prevents virus-induced increases in Ciliated and goblet Cells whereas IL-13 blockade (using s-IL-13Rα2-Fc) exacerbates Ciliated Cell hyperplasia but still inhibits goblet Cell metaplasia. The distinct effects of EGFR and IL-13 inhibitors after viral reprogramming suggest that these combined therapeutic strategies may also correct epithelial architecture in the setting of airway inflammatory disorders characterized by a similar pattern of chronic EGFR activation, IL-13 expression, and Ciliated-to-goblet Cell metaplasia.
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role of f box factor foxj1 in differentiation of Ciliated airway epithelial Cells
American Journal of Physiology-lung Cellular and Molecular Physiology, 2004Co-Authors: Tao Huang, Edward J Richer, Jenserik Harboe Schmidt, Joseph Zabner, Zea Borok, Steven L. BrodyAbstract:Factors required for commitment of an undifferentiated airway epithelial Cell to a Ciliated Cell are unknown. Cell ultrastructure analysis indicates Ciliated Cell commitment activates a multistage ...
Rebecca A. Wingert - One of the best experts on this subject based on the ideXlab platform.
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ppargc1a controls Ciliated Cell development by regulating prostaglandin biosynthesis
Social Science Research Network, 2020Co-Authors: Joseph M. Chambers, Amanda Addiego, Rebecca A. WingertAbstract:Cilia are microtubule-based organelles that function in a multitude of physiological contexts to perform chemosensing, mechanosensing, or fluid propulsion. The process of ciliogenesis is highly regulated and disruptions result in disease states termed ciliopathies. Here, we show novel roles for peroxisome proliferator-activated receptor gamma 1 alpha ( ppargc1a ) during ciliogenesis in nodal, mono-, and multiCiliated Cells (MCCs) as well as discernment of renal tubule Ciliated Cell fate during embryogenesis. We discovered that ppargc1a performs both roles by affecting prostaglandin signaling, where cilia formation and renal MCC fate were restored with prostaglandin E 2 (PGE 2 ) treatment. Genetic disruption of ppargc1a specifically reduced expression of the prostanoid biosynthesis gene prostaglandin-endoperoxide synthase 1 ( ptgs1 ), and suboptimal knockdown of both genes revealed a synergistic effect. Further, ptgs1 overexpression rescued ciliogenesis and renal MCCs in ppargc1a deficient embryos. These findings position Ppargc1a as an essential genetic regulator of prostaglandin signaling during Ciliated Cell ontogeny.
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Ppargc1a Controls Ciliated Cell Development by Regulating Prostaglandin Biosynthesis.
Cell Reports, 2020Co-Authors: Joseph M. Chambers, Amanda Addiego, Ana L Flores-mireles, Rebecca A. WingertAbstract:Cilia are microtubule-based organelles that function in a multitude of physiological contexts to perform chemosensing, mechanosensing, and fluid propulsion. The process of ciliogenesis is highly regulated, and disruptions result in disease states termed ciliopathies. Here, we report that peroxisome proliferator-activated receptor gamma, coactivator 1 alpha (ppargc1a) is essential for ciliogenesis in nodal, mono-, and multiCiliated Cells (MCCs) and for discernment of renal tubule Ciliated Cell fate during embryogenesis. ppargc1a performs these functions by affecting prostaglandin signaling, whereby cilia formation and renal MCC fate are restored with prostaglandin E2 (PGE2) treatment in ppargc1a-deficient animals. Genetic disruption of ppargc1a specifically reduces expression of the prostanoid biosynthesis gene prostaglandin-endoperoxide synthase 1 (ptgs1), and suboptimal knockdown of both genes shows this synergistic effect. Furthermore, ptgs1 overexpression rescues ciliogenesis and renal MCCs in ppargc1a-deficient embryos. These findings position Ppargc1a as a key genetic regulator of prostaglandin signaling during Ciliated Cell ontogeny.
Nevis Fregien - One of the best experts on this subject based on the ideXlab platform.
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submersion and hypoxia inhibit Ciliated Cell differentiation in a notch dependent manner
American Journal of Respiratory Cell and Molecular Biology, 2014Co-Authors: Benjamin J Gerovac, Gregory E Conner, Matthias Salathe, Monica Valencia, Nathalie Baumlin, Nevis FregienAbstract:The epithelium that lines the conducting airways is composed of several distinct Cell types that differentiate from common progenitor Cells. The signals that control fate selection and differentiation of Ciliated Cells, a major component of the epithelium, are not completely understood. Ciliated Cell differentiation can be accomplished in vitro when primary normal human bronchial epithelial (NHBE) Cells are cultured at an air–liquid interface, but is inhibited when NHBE Cells are cultured under submerged conditions. The mechanism by which submersion prevents ciliogenesis is not understood, but may provide clues to in vivo regulation of Ciliated Cell differentiation. We hypothesized that submersion creates a hypoxic environment that prevents Ciliated Cell differentiation by blocking the gene expression program required for ciliogenesis. This was confirmed by showing that expression of multicilin and Forkhead box J1, key factors needed for Ciliated Cell differentiation, was inhibited when NHBE Cells were cultured in submerged and hypoxic conditions. Multicilin and Forkhead box J1 expression and Ciliated Cell differentiation were restored in submerged and hypoxic Cells upon treatment with the γ-secretase inhibitor, N-[(3,5-difluorophenyl)acetyl]-L-alanyl-2-phenyl]glycine-1,1-dimethylethyl ester (DAPT), which suggested that Notch signaling was involved. Overexpression of Notch intraCellular domain inhibited differentiation in the presence of DAPT, confirming the role of Notch signaling. These results indicate that submersion and hypoxia prevent Ciliated Cell differentiation by maintaining Notch signaling, which represses genes necessary for ciliogenesis. These data provide new insights into the molecular mechanisms that control human bronchial differentiation.
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decreased soluble adenylyl cyclase activity in cystic fibrosis is related to defective apical bicarbonate exchange and affects ciliary beat frequency regulation
Journal of Biological Chemistry, 2010Co-Authors: Andreas Schmid, Nathalie Schmid, Gregory E Conner, Pedro Ivonnet, Zoltan Sutto, Gabor Horvath, Lisa Novak, Nevis FregienAbstract:Human airway cilia contain soluble adenylyl cyclase (sAC) that produces cAMP upon HCO3−/CO2 stimulation to increase ciliary beat frequency (CBF). Because apical HCO3− exchange depends on cystic fibrosis transmembrane conductance regulator (CFTR), malfunctioning CFTR might impair sAC-mediated CBF regulation in Cells from patients with cystic fibrosis (CF). By Western blot, sAC isoforms are equally expressed in normal and CF airway epithelial Cells, but CBF decreased more in CF than normal Cells upon increased apical HCO3−/CO2 exposure in part because of greater intraCellular acidification from unbalanced CO2 influx (estimated by 2′,7′-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF) fluorescence). Importantly, Ciliated Cell-specific cAMP production (estimated by FRET fluorescence ratio changes of tagged cAMP-dependent protein kinase (PKA) subunits expressed under a Ciliated Cell-specific promoter) in response to increased apical HCO3−/CO2 perfusion was higher in normal compared with CF Cells. Inhibition of bicarbonate influx via CFTR (CFTRinh172) and inhibition of sAC (KH7) and PKA activation (H89) led to larger CBF declines in normal Cells, now comparable with changes seen in CF Cells. These inhibitors also reduced FRET changes in normal Cells to the level of CF Cells with the expected exception of H89, which does not prevent dissociation of the fluorescently tagged PKA subunits. Basolateral permeabilization and subsequent perfusion with HCO3−/CO2 rescued CBF and FRET changes in CF Cells to the level of normal Cells. These results suggest that CBF regulation by sAC-produced cAMP could be impaired in CF, thereby possibly contributing to mucociliary dysfunction in this disease, at least during disease exacerbations when airway acidification is common.
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real time analysis of camp mediated regulation of ciliary motility in single primary human airway epithelial Cells
Journal of Cell Science, 2006Co-Authors: Andreas Schmid, Ge Bai, Nathalie Schmid, Manuela Zaccolo, Lawrence E Ostrowski, Gregory E Conner, Nevis Fregien, Matthias SalatheAbstract:Airway ciliary beat frequency regulation is complex but in part influenced by cyclic adenosine monophosphate (cAMP)-mediated changes in cAMP-dependent kinase activity, yet the cAMP concentration required for increases in ciliary beat frequency and the temporal relationship between ciliary beat frequency and cAMP changes are unknown. A lentiviral gene transfer system was developed to express a fluorescence resonance energy transfer (FRET)-based cAMP sensor in Ciliated Cells. Expression of fluorescently tagged cAMP-dependent kinase subunits from the Ciliated-Cell-specific foxj1 promoter enhanced expression in fully differentiated Ciliated human airway epithelial Cells, and permitted simultaneous measurements of ciliary beat frequency and cAMP (represented by the FRET ratio). Apical application of forskolin (1 μM, 10 μM, 20 μM) and, in permeabilized Cells, basolateral cAMP (20 μM, 50 μM, 100 μM) caused dose-dependent, albeit similar and simultaneous–increases in cAMP and ciliary beat frequency. However, decreases in cAMP preceded decreases in ciliary beat frequency, suggesting that either Cellular cAMP decreases before ciliary cAMP or the dephosphorylation of target proteins by phosphatases occur at a rate slower than the rate of cAMP hydrolysis.