Coupled Nucleotide and Mucin Hypersecretion from Goblet-Cell Metaplastic Human Airway Epithelium

Okada, Seiko F.; Zhang, Liqun; Kreda, Silvia M.; Abdullah, Lubna H.; Davis, C. William; Pickles, Raymond J.; Lazarowski, Eduardo R.; Boucher, Richard C.

doi:10.1165/rcmb.2010-0253oc

Cited by 56 publications

(56 citation statements)

References 50 publications

(72 reference statements)

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“…Because Entpd7 was highly expressed in goblet cells, as well as absorptive ECs, it is possible that Entpd7 is expressed in the membrane of mucin-containing vacuoles of goblet cells to control ATP levels in the vacuole. Given that goblet cells of the airway were shown to secrete ATP, as well as mucin (42), intestinal goblet cells might be a major source of luminal ATP, the level of which is closely regulated by ENTPDase7.…”

Section: Discussionmentioning

confidence: 99%

Ecto-Nucleoside Triphosphate Diphosphohydrolase 7 Controls Th17 Cell Responses through Regulation of Luminal ATP in the Small Intestine

Kusu

Kayama

Kinoshita

et al. 2013

The Journal of Immunology

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Extracellular ATP is released from live cells in controlled conditions, as well as dying cells in inflammatory conditions, and, thereby, regulates T cell responses, including Th17 cell induction. The level of extracellular ATP is closely regulated by ATP hydrolyzing enzymes, such as ecto-nucleoside triphosphate diphosphohydrolases (ENTPDases). ENTPDase1/CD39, which is expressed in immune cells, was shown to regulate immune responses by downregulating the ATP level. In this study, we analyzed the immunomodulatory function of ENTPDase7, which is preferentially expressed in epithelial cells in the small intestine. The targeted deletion of Entpd7 encoding ENTPDase7 in mice resulted in increased ATP levels in the small intestinal lumen. The number of Th17 cells was selectively increased in the small intestinal lamina propria in Entpd7−/− mice. Th17 cells were decreased by oral administration of antibiotics or the ATP antagonist in Entpd7−/− mice, indicating that commensal microbiota-dependent ATP release mediates the enhanced Th17 cell development in the small intestinal lamina propria of Entpd7−/− mice. In accordance with the increased number of small intestinal Th17 cells, Entpd7−/− mice were resistant to oral infection with Citrobacter rodentium. Entpd7−/− mice suffered from severe experimental autoimmune encephalomyelitis, which was associated with increased numbers of CD4+ T cells producing both IL-17 and IFN-γ. Taken together, these findings demonstrate that ENTPDase7 controls the luminal ATP level and, thereby, regulates Th17 cell development in the small intestine.

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Section: Discussionmentioning

confidence: 99%

Ecto-Nucleoside Triphosphate Diphosphohydrolase 7 Controls Th17 Cell Responses through Regulation of Luminal ATP in the Small Intestine

Kusu

Kayama

Kinoshita

et al. 2013

The Journal of Immunology

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“…At lower concentrations, the H 2 O 2 stimulation of CFTR to increase mucociliary clearance occurs concomitantly with increases in CBF (2) through Ca 21 -mediated and cAMP-mediated stimulation. In addition, mucins are also likely secreted through a Ca 21 -mediated mechanism (48). It is interesting to speculate that H 2 O 2 production by epithelial cells contributes to the normal ASL volume increases that accompany the Ca 21 -mediated stimulation of CBF and mucin secretions in the airways.…”

Section: Discussionmentioning

confidence: 99%

H₂O₂ Stimulates Cystic Fibrosis Transmembrane Conductance Regulator through an Autocrine Prostaglandin Pathway, Using Multidrug-Resistant Protein–4

Conner

Ivonnet²,

Gelin³

et al. 2013

Am J Respir Cell Mol Biol

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Cystic fibrosis transmembrane conductance regulator (CFTR) activity is essential for the maintenance of airway surface liquid depth, and therefore mucociliary clearance. Reactive oxygen species, increased during inflammatory airway diseases, alter CFTR activity. Here, H 2 O 2 levels in the surface liquid of normal human bronchial epithelial cultures differentiated at the air-liquid interface were estimated, and H 2 O 2 -mediated changes in CFTR activity were examined. In Ussing chambers, H 2 O 2 -induced anion currents were sensitive to the CFTR inhibitors CFTR inh 172 and GlyH-101. These currents were absent in cells from patients with cystic fibrosis. Effective airway clearance relies on the regulation of ciliary beat frequency (CBF), adequate airway surface liquid (ASL) volume, and correct mucus properties. Mucus hydration and thus the viscosity of mucus depend, in part, on the movement of water through the airway epithelium to the mucosal surface via the coordinated activities of ion channels. An important player in this regulation is the cystic fibrosis transmembrane conductance regulator (CFTR) that passes chloride (and other anions) while regulating the epithelial Na 1 channel to balance osmolarity (1). CFTR and CBF are both regulated through increases in cyclic adenosine monophosphate (cAMP) and protein kinase A activity (2-4). Thus, mediators that stimulate adenylyl cyclase are expected to result in improved airway clearance by increasing chloride secretion and CBF.Airway epithelia actively synthesize H 2 O 2 via the nicotinamide adenine dinucleotide phosphate-reduced (NADPH) dual oxidases Duox1 and Duox2, as reviewed by Fischer (5), presumably for the use of the lactoperoxidase host defense against infection (6-9). Duox1 and Duox2 are regulated by cytokines (10-12) and bacterial products (10,13,14) to increase H 2 O 2 synthesis to high levels (10). Stimulation with interferon-g increased H 2 O 2 synthesis levels 30-fold when measured in air-liquid interface (ALI) cultures of normal human airway epithelial cells (10), leading to 10 mM H 2 O 2 in washes used for assays. The experiments presented here address ASL H 2 O 2 and show that it is present at high concentrations.In addition to its role in stimulating lactoperoxidase host defense, H 2 O 2 may also regulate the innate defense related to mucociliary clearance. In the bronchial submucosal adenocarcinoma cell line Calu-3, CFTR activity is increased by exposure of the airway epithelium to exogenous H 2 O 2 (15, 16). In cell lines, the effect of H 2 O 2 on CFTR is reported to occur through increases in transmembrane adenylyl cyclase activity and through the activation of two K 1 channels (15), most likely KCNQ1 and KCNN4, which are located in the basolateral membrane. These channels are believed to provide a driving force for apical Cl 2 release (17, 18). Yet the mechanisms by which H 2 O 2 leads to the activation of the cAMP pathway are not understood. Although both Calu-3 cells (19) and bovine tracheal epithelia (20) have been shown to respond to isop...

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“…For example, we recently demonstrated that plasma membrane pannexin 1 largely contributes to conductive ATP release from normal airways dominated by ciliated cells (22). Deletion of the pannexin 1 gene resulted in impaired ATP release in mouse tracheas ex vivo (22).We also demonstrated that mucin granules are an important additional source of nucleotide release (8,9,18). That is, primary cultures of human airway epithelial cells that were induced to develop goblet cell hyperplasia via respiratory syncytial virus infection or IL-13 exposure acquired enhanced ATP release from the secretory pathway that correlated with increased mucin secretion and was not affected by pannexin inhibitors (18).…”

mentioning

confidence: 82%

Vesicular nucleotide transporter regulates the nucleotide content in airway epithelial mucin granules

Sesma

Kreda

Okada

et al. 2013

American Journal of Physiology-Cell Physiology

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nucleotide transporter regulates the nucleotide content in airway epithelial mucin granules. Am J Physiol Cell Physiol 304: C976 -C984, 2013. First published March 6, 2013; doi:10.1152/ajpcell.00371.2012.-Nucleotides within the airway surface liquid promote fluid secretion via activation of airway epithelial purinergic receptors. ATP is stored within and released from mucin granules as co-cargo with mucins, but the mechanism by which ATP, and potentially other nucleotides, enter the lumen of mucin granules is not known. We assessed the contribution of the recently identified SLC17A9 vesicle nucleotide transporter (VNUT) to the nucleotide availability within isolated mucin granules and further examined the involvement of VNUT in mucin granule secretion-associated nucleotide release. RT-PCR and Western blot analyses indicated that VNUT is abundantly expressed in airway epithelial goblet-like Calu-3 cells, migrating as a duplex with apparent mobility of 55 and 60 kDa. Subcellular fractionation studies indicated that VNUT55 was associated with high-density mucin granules, whereas VNUT60 was associated with low-density organelles. Immunofluorescence studies showed that recombinant VNUT localized to mucin granules and other organelles. Mucin granules isolated from VNUT short hairpin RNA-expressing cells exhibited a marked reduction of ATP, ADP, AMP, and UTP levels within granules. Ca 2ϩ -regulated vesicular ATP release was markedly reduced in these cells, but mucin secretion was not affected. These results suggest that VNUT is the relevant nucleotide transporter responsible for the uptake of cytosolic nucleotides into mucin granules. By controlling the entry of nucleotides into mucin granules, VNUT contributes to the release of purinergic signaling molecules necessary for the proper hydration of co-released mucins.VNUT; SLC17A9; mucin granules; ATP release; mucin secretion EXTRACELLULAR NUCLEOTIDES are key components of the mucociliary clearance process that traps and removes inhaled particles and microorganisms from the lung. Nucleotides/nucleosides within the airway surface liquid (ASL) promote mucin secretion via activation of P2Y 2 receptors in mucous (goblet) cells and stimulate fluid secretion/mucin hydration via A 2B and P2Y 2 receptor activation in ciliated cells (11). Deficient fluid secretion results in abnormal mucous hydration/clearance, leading to lung failure in cystic fibrosis, the most prevalent potentially lethal genetic disease in the United States and a major factor contributing to the progressive airway obstruction associated with chronic obstructive lung disease (4).Despite the importance of ASL nucleotides in airway physiology, the mechanisms by which airway epithelial cells release nucleotides have just begun to be addressed. For example, we recently demonstrated that plasma membrane pannexin 1 largely contributes to conductive ATP release from normal airways dominated by ciliated cells (22). Deletion of the pannexin 1 gene resulted in impaired ATP release in mouse tracheas ex vivo (22).We also ...

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Coupled Nucleotide and Mucin Hypersecretion from Goblet-Cell Metaplastic Human Airway Epithelium

Cited by 56 publications

References 50 publications

Ecto-Nucleoside Triphosphate Diphosphohydrolase 7 Controls Th17 Cell Responses through Regulation of Luminal ATP in the Small Intestine

Ecto-Nucleoside Triphosphate Diphosphohydrolase 7 Controls Th17 Cell Responses through Regulation of Luminal ATP in the Small Intestine

H₂O₂ Stimulates Cystic Fibrosis Transmembrane Conductance Regulator through an Autocrine Prostaglandin Pathway, Using Multidrug-Resistant Protein–4

Vesicular nucleotide transporter regulates the nucleotide content in airway epithelial mucin granules

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Coupled Nucleotide and Mucin Hypersecretion from Goblet-Cell Metaplastic Human Airway Epithelium

Cited by 56 publications

References 50 publications

Ecto-Nucleoside Triphosphate Diphosphohydrolase 7 Controls Th17 Cell Responses through Regulation of Luminal ATP in the Small Intestine

Ecto-Nucleoside Triphosphate Diphosphohydrolase 7 Controls Th17 Cell Responses through Regulation of Luminal ATP in the Small Intestine

H2O2 Stimulates Cystic Fibrosis Transmembrane Conductance Regulator through an Autocrine Prostaglandin Pathway, Using Multidrug-Resistant Protein–4

Vesicular nucleotide transporter regulates the nucleotide content in airway epithelial mucin granules

Contact Info

Product

Resources

About

H₂O₂ Stimulates Cystic Fibrosis Transmembrane Conductance Regulator through an Autocrine Prostaglandin Pathway, Using Multidrug-Resistant Protein–4