NEU1 Sialidase Expressed in Human Airway Epithelia Regulates Epidermal Growth Factor Receptor (EGFR) and MUC1 Protein Signaling

Lillehoj, Erik P.; Hyun, Sang Won; Feng, Chiguang; Zhang, Lei; Liu, Anguo; Guang, Wei; Nguyen, Chinh; Luzina, Irina G.; Atamas, Sergei P.; Passaniti, Antonino; Twaddell, William S.; Puché, Adam C.; Wang, Lai-Xi; Cross, Alan S.; Goldblum, Simeon E.

doi:10.1074/jbc.m111.292888

Cited by 72 publications

(165 citation statements)

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“…AGS cells in 24-well plates were transfected with 20 pmoles/well of a MUC1-targeting small interfering RNA (siRNA) (sense, 5=-GUUCAGUGCCCAGCUCUACdTdT-3=; antisense, 5=-GUAGAGCUGGGCACUGAACdTdT-3=) or with a nontargeting control siRNA (sense, 5=-GCGCGCUUUGUAGGAUUCGdTdT-3=; antisense, 5=-CGAAUCCUACAAAGCGCGCdTdT-3=) (Dharmacon, Lafayette, CO), using 2.0 l/well of Lipofectamine (Invitrogen) as described (18). Prior studies have documented the specificity of the MUC1 siRNA with no demonstrable off-target effects (18,23,30,32,33,35).…”

Section: Methodsmentioning

confidence: 99%

Suppression of IL-8 production in gastric epithelial cells by MUC1 mucin and peroxisome proliferator-associated receptor-γ

Park

Guang

Blanchard

et al. 2012

American Journal of Physiology-Gastrointestinal and Liver Physi

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MUC1 is a membrane-tethered mucin expressed on the apical surface of epithelial cells. Our previous report (Guang W, Ding H, Czinn SJ, Kim KC, Blanchard TG, Lillehoj EP. J Biol Chem 285: 20547–20557, 2010) demonstrated that expression of MUC1 in AGS gastric epithelial cells limits Helicobacter pylori infection and reduces bacterial-driven IL-8 production. In this study, we identified the peroxisome proliferator-associated receptor-γ (PPARγ) upstream of MUC1 in the anti-inflammatory pathway suppressing H. pylori- and phorbol 12-myristate 13-acetate (PMA)-stimulated IL-8 production. Treatment of AGS cells with H. pylori or PMA increased IL-8 levels in cell culture supernatants compared with cells treated with the respective vehicle controls. Prior small interfering (si)RNA-induced MUC1 silencing further increased H. pylori - and PMA-stimulated IL-8 levels compared with a negative control siRNA. MUC1-expressing AGS cells pretreated with the PPARγ agonist troglitazone (TGN) had reduced H. pylori - and PMA-stimulated IL-8 levels compared with cells treated with H. pylori or PMA alone. However, following MUC1 siRNA knockdown, no differences in IL-8 levels were seen between TGN/ H. pylori and H. pylori -only cells or between TGN/PMA and PMA-only cells. Finally, TGN-treated AGS cells had increased Muc1 promoter activity, as measured using a Muc1-luciferase reporter gene, and greater MUC1 protein levels by Western blot analysis, compared with vehicle controls. These results support the hypothesis that PPARγ stimulates MUC1 expression by AGS cells, thereby attenuating H. pylori - and PMA-induced IL-8 production.

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Section: Methodsmentioning

confidence: 99%

Suppression of IL-8 production in gastric epithelial cells by MUC1 mucin and peroxisome proliferator-associated receptor-γ

Park

Guang

Blanchard

et al. 2012

American Journal of Physiology-Gastrointestinal and Liver Physi

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“…NEU3 was the second most abundant EC sialidase found at the mRNA level. When airway epithelia were assayed for sialidase activity for ganglioside substrates, SA release was evident (31), suggesting that the Ϸ30% residual sialidase activity detected in airway epithelia after NEU1 silencing might be explained, in part, through NEU3 enzymatic activity. Accordingly, in the present study we surveyed a diverse collection of human airway ECs for sialidase activity against ganglioside substrates, identified NEU3 mRNA and protein in these same epithelia, as well as in intact human airway tissues, and correlated siRNA-induced knockdown of NEU3 expression with reduced ganglioside-directed sialidase activity.…”

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confidence: 99%

“…In recent studies in human airway epithelia, we found heat-labile sialidase activity for the fluorogenic substrate, 2=-(4-methylumbelliferyl)-␣-D-Neu5Ac (4-MU-Neu5Ac), that was dose dependently inhibited by the sialidase inhibitor, 2-deoxy-Neu5Ac, but not its negative control, 2-keto-3-deoxyoctulosonic acid (31). These airway epithelia expressed predominantly NEU1 sialidase at the mRNA and protein levels, and small interfering (si)RNA-induced silencing of NEU1 diminished sialidase activity for the 4-MUNeu5Ac substrate Ͼ70% compared with control siRNA-transfected ECs.…”

mentioning

confidence: 99%

“…The human airway epithelial surface is decorated with glycoconjugates expressing SA connected to underlying sugars in ␣-2,3 and ␣-2,6 linkages (12). These sialylated glycoconjugates include specific receptors that respond to danger signals (9), serve as binding sites for pathogenic bacteria and viruses (23,31,33), and initiate epithelial repair programs (9). Superimposed on this highly sialylated EC surface is a constantly moving, SA-rich mucous blanket (50).…”

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confidence: 99%

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Human airway epithelia express catalytically active NEU3 sialidase

Lillehoj

Hyun²,

Feng

et al. 2014

American Journal of Physiology-Lung Cellular and Molecular Phys

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acids on glycoconjugates play a pivotal role in many biological processes. In the airways, sialylated glycoproteins and glycolipids are strategically positioned on the plasma membranes of epithelia to regulate receptor-ligand, cell-cell, and host-pathogen interactions at the molecular level. We now demonstrate, for the first time, sialidase activity for ganglioside substrates in human airway epithelia. Of the four known mammalian sialidases, NEU3 has a substrate preference for gangliosides and is expressed at mRNA and protein levels at comparable abundance in epithelia derived from human trachea, bronchi, small airways, and alveoli. In small airway and alveolar epithelia, NEU3 protein was immunolocalized to the plasma membrane, cytosolic, and nuclear subcellular fractions. Small interfering RNA-induced silencing of NEU3 expression diminished sialidase activity for a ganglioside substrate by Ͼ70%. NEU3 immunostaining of intact human lung tissue could be localized to the superficial epithelia, including the ciliated brush border, as well as to nuclei. However, NEU3 was reduced in subepithelial tissues. These results indicate that human airway epithelia express catalytically active NEU3 sialidase.

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“…However, Lillehoj et al reported sialidase activity in human airway epithelial cells derived from lysosomal neuraminidase (NEU1) (57). Clostridium perfringens neuraminidase, which is highly homologous to NEU1 (58), was unable to support SIV/606 replication in vitro (data not shown), possibly because the neuraminidase activity of Clostridium perfringens has optimal pH and calcium ion requirements different from those of Vibrio cholerae (59).…”

Section: Discussionmentioning

confidence: 99%

An Eight-Segment Swine Influenza Virus Harboring H1 and H3 Hemagglutinins Is Attenuated and Protective against H1N1 and H3N2 Subtypes in Pigs

Mašić

Pyo

Babiuk

et al. 2013

J Virol

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Swine influenza virus (SIV) infections continue to cause production losses in the agricultural industry in addition to being a human public health concern. The primary method of controlling SIV is through vaccination. The killed SIV vaccines currently in use must be closely matched to the challenge virus, and their protective efficacy is limited. Live attenuated influenza vaccines (LAIV) provide strong, long-lived cell-mediated and humoral immunity against different influenza virus subtypes with no need for antigen matching. Here we report the generation of a new potential LAIV, an eight-segment SIV harboring two different SIV hemagglutinins (HAs), H1 and H3, in the genetic background of H1N1 SIV. This mutant SIV was generated by fusing the H3 HA ectodomain from A/Swine/Texas/4199-2/98 (H3N2) to the cytoplasmic tail, transmembrane domain, and stalk region of neuraminidase (NA) from A/Swine/Saskatchewan/18789/02 (H1N1) SIV. While this H1-H3 chimeric SIV, when propagated in vitro in the presence of exogenous neuraminidase, showed kinetics and growth properties similar to those of the parental wild-type virus, in vivo it was highly attenuated in pigs, demonstrating a great potential for serving as a dual LAIV. Furthermore, vaccination with the H1-H3 virus elicited robust immune responses, which conferred complete protection against infections with both H1 and H3 SIV subtypes in pigs.

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NEU1 Sialidase Expressed in Human Airway Epithelia Regulates Epidermal Growth Factor Receptor (EGFR) and MUC1 Protein Signaling

Cited by 72 publications

References 79 publications

Suppression of IL-8 production in gastric epithelial cells by MUC1 mucin and peroxisome proliferator-associated receptor-γ

Suppression of IL-8 production in gastric epithelial cells by MUC1 mucin and peroxisome proliferator-associated receptor-γ

Human airway epithelia express catalytically active NEU3 sialidase

An Eight-Segment Swine Influenza Virus Harboring H1 and H3 Hemagglutinins Is Attenuated and Protective against H1N1 and H3N2 Subtypes in Pigs

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