1. The NEU1-selective sialidase inhibitor, C9-butyl-amide-DANA, blocks sialidase activity and NEU1-mediated bioactivities in human lung in vitro and murine lung in vivo.
- Author
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Hyun SW, Liu A, Liu Z, Cross AS, Verceles AC, Magesh S, Kommagalla Y, Kona C, Ando H, Luzina IG, Atamas SP, Piepenbrink KH, Sundberg EJ, Guang W, Ishida H, Lillehoj EP, and Goldblum SE
- Subjects
- Animals, Bacterial Proteins genetics, Bacterial Proteins metabolism, Cathepsin A genetics, Cathepsin A metabolism, Cell Movement drug effects, Endothelial Cells cytology, Endothelial Cells drug effects, Endothelial Cells enzymology, Endothelium, Vascular cytology, Endothelium, Vascular drug effects, Endothelium, Vascular enzymology, Epithelial Cells cytology, Epithelial Cells drug effects, Epithelial Cells enzymology, Fibroblasts cytology, Fibroblasts drug effects, Fibroblasts enzymology, Flagellin antagonists & inhibitors, Flagellin pharmacology, Gene Expression Regulation, Humans, Hydrolysis, Isoenzymes antagonists & inhibitors, Isoenzymes genetics, Isoenzymes metabolism, Lung cytology, Lung enzymology, Mice, Models, Molecular, Mucin-1 genetics, Mucin-1 metabolism, N-Acetylneuraminic Acid analogs & derivatives, N-Acetylneuraminic Acid chemistry, Neuraminidase genetics, Neuraminidase metabolism, Protein Binding, Protein Conformation, alpha-Helical, Protein Conformation, beta-Strand, Protein Domains, Protein Interaction Domains and Motifs, Pseudomonas aeruginosa chemistry, Enzyme Inhibitors pharmacology, Lung drug effects, Mucin-1 chemistry, N-Acetylneuraminic Acid pharmacology, Neuraminidase antagonists & inhibitors
- Abstract
Neuraminidase-1 (NEU1) is the predominant sialidase expressed in human airway epithelia and lung microvascular endothelia where it mediates multiple biological processes. We tested whether the NEU1-selective sialidase inhibitor, C9-butyl-amide-2-deoxy-2,3-dehydro-N-acetylneuraminic acid (C9-BA-DANA), inhibits one or more established NEU1-mediated bioactivities in human lung cells. We established the IC50 values of C9-BA-DANA for total sialidase activity in human airway epithelia, lung microvascular endothelia and lung fibroblasts to be 3.74 µM, 13.0 µM and 4.82 µM, respectively. In human airway epithelia, C9-BA-DANA dose-dependently inhibited flagellin-induced, NEU1-mediated mucin-1 ectodomain desialylation, adhesiveness for Pseudomonas aeruginosa and shedding. In lung microvascular endothelia, C9-BA-DANA reversed NEU1-driven restraint of cell migration into a wound and disruption of capillary-like tube formation. NEU1 and its chaperone/transport protein, protective protein/cathepsin A (PPCA), were differentially expressed in these same cells. Normalized NEU1 protein expression correlated with total sialidase activity whereas PPCA expression did not. In contrast to eukaryotic sialidases, C9-BA-DANA exerted far less inhibitory activity for three selected bacterial neuraminidases (IC50 > 800 µM). Structural modeling of the four human sialidases and three bacterial neuraminidases revealed a loop between the seventh and eighth strands of the β-propeller fold, that in NEU1, was substantially shorter than that seen in the six other enzymes. Predicted steric hindrance between this loop and C9-BA-DANA could explain its selectivity for NEU1. Finally, pretreatment of mice with C9-BA-DANA completely protected against flagellin-induced increases in lung sialidase activity. Our combined data indicate that C9-BA-DANA inhibits endogenous and ectopically expressed sialidase activity and established NEU1-mediated bioactivities in human airway epithelia, lung microvascular endothelia, and fibroblasts in vitro and murine lungs in vivo., (© The Author 2016. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2016
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