Abstract
Mucus hypersecretion is a crucial feature of pulmonary diseases such as asthma, chronic bronchitis and cystic fibrosis. Despite much research, there is still no effective therapy for this condition. Recently, we showed that the myristoylated, alanine-rich C-kinase substrate (MARCKS) protein is required for mucus secretion by human bronchial epithelial cells in culture1. Having synthesized a peptide corresponding to the N-terminal domain of MARCKS, we now show that the intratracheal instillation of this peptide blocks mucus hypersecretion in a mouse model of asthma. A missense peptide with the same amino acid composition has no effect. Based on quantitative histochemical analysis of the mouse airways, the peptide seems to act by blocking mucus release from goblet cells, possibly by inhibiting the attachment of MARCKS to membranes of intracellular mucin granules. These results support a pivotal role for MARCKS protein, specifically its N-terminal region, in modulating this secretory process in mammalian airways. Intratracheal administration of this MARCKS-related peptide could therapeutically reduce mucus secretion in the airways of human patients with asthma, chronic bronchitis and cystic fibrosis.
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References
Li, Y., Martin, L.D., Spizz, G. & Adler, K.B. MARCKS protein is a key molecule regulating mucin secretion by human airway epithelial cells in vitro. J. Biol. Chem. 276, 40982–40990 (2001).
Eum, S.Y., Haile, S., Huerre, M. & Vargaftig, B.B. Eosinophil recruitment into the respiratory epithelium following antigenic challenge in hyper-IgE mice is accompanied by interleukin-5-dependent bronchial hyperresponsiveness. Proc. Natl. Acad. Sci. USA 92, 12290–12294 (1995).
Haile, S. et al. Mucous cell metaplasia and inflammatory cell recruitment are dissociated in allergic mice following antibody- and drug-dependent cell depletion in a murine model of asthma. Am. J. Respir. Cell Mol. Biol. 20, 891–902 (1999).
Singer, M., Lefort, J. & Vargaftig, B.B. Granulocyte depletion and dexamethasone differentially modulate airway hyperreactivity, inflammation, mucus accumulation, and secretion induced by rmIL-13 or antigen. Am. J. Respir. Cell Mol. Biol. 26, 74–84 (2002).
Vargaftig, B.B. & Singer, M. Leukotrienes, IL-13, and chemokines cooperate to induce BHR and mucus in allergic mouse lungs. Am. J. Physiol. Lung Cell. Mol. Physiol. 284, L260–L269 (2003).
Stumpo, D.J., Bock, C.B., Tuttle, J.S. & Blackshear, P.J. MARCKS deficiency in mice leads to abnormal brain development and perinatal death. Proc. Nat. Acad. Sci. USA 92, 944–948 (1995).
Rose, S.D., Lejen, T., Zhang, L. & Trifaro, J.M. Chromaffin cell F-actin disassembly and potentiation of catecholamine release in response to protein kinase C activation by phorbol esters is mediated through myristoylated alanine-rich C kinase substrate phosphorylation. J. Biol. Chem. 276, 36757–36763 (2001).
Elzagallaai, A., Rose, S.D. & Trifaro, J.M. Platelet secretion induced by phorbol ester stimulation is mediated through phosphorylation of MARCKS: a MARCKS-derived peptide blocks MARCKS phosphorylation and serotonin release without affecting pleckstrin phosphorylation. Blood 95, 894–902 (2000).
Vaughan, P.F., Walker, J.H. & Peers, C. The regulation of neurotransmitter secretion by protein kinase C. J. Mol. Neurobiol. 18, 125–155 (1998).
Trifaro, J., Rose, S.D., Lejen, T. & Elzagallaai, A. Two pathways control chromaffin cell cortical F-actin dynamics during exocytosis. Biochimie 82, 339–352 (2000).
Salli, U., Supanic, S. & Stormshak, F. Phosphorylation of myristoylated alanine rich C kinase substrate (MARCKS) protein is associated with bovine luteal oxytocin exocytosis. Biol. Reprod. 63, 12–20 (2000).
Hartness, M.E., Wade, J.A., Walker, J.H. & Vaughan, P.F. Overexpression of the myristoylated alanine-rich C kinase substrate decreases uptake and K+-evoked release of noradrenaline in the human neuroblastoma SH-SY5Y. Eur. J. Neurosci. 13, 925–934 (2001).
Seykora, J.T., Myatt, M.M., Allen, L.A.H., Ravetch, J.V. & Aderem, A. Molecular determinants of the myristoyl-electrostatic switch of MARCKS. J. Biol. Chem. 271, 18797–18802 (1996).
Spizz, G. & Blackshear, P.J. Identification and characterization of cathepsin B as the cellular MARCKS cleaving enzyme. J. Biol. Chem. 272, 23833–23842 (1997).
Shekels, L.L. et al. Mouse gastric mucin: cloning and chromosomal localization. Biochem. J. 311, 775–785 (1995).
Vargaftig, B.B. & Singer, M. Leukotrienes mediate part of Ova-induced lung effects in mice via EGFR. Am. J. Physiol. Lung Cell Mol. Physiol. 285, L808–L818 (2003).
Wu, T.T. & Castle, J.D. Evidence for colocalization and interaction between 37 and 39 kDa isoforms of secretory carrier membrane proteins (SCAMPs). J. Cell Sci. 110, 1533–1541 (1997).
Leverkoehne, I. & Gruber, A.D. The murine mCLCA3(alias gob-5) protein is located in the mucin granule membranes of intestinal, respiratory and uterine goblet cells. J. Histochem. Cytochem. 50, 829–838 (2002).
Acknowledgements
This study was supported by grant RO1-HL36982 from the National Institutes of Health (to K.B.A.). The authors thank P. Blackshear (National Institute of Environmental Health Sciences) for providing invaluable assistance and reagents, and S. Ho (University of Minnesota) for generously providing antibodies.
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L.D.M., Y.L. and K.B.A. are listed on a patent filed by North Carolina State University, describing the effects of the peptides used in these studies. The licensed technology is held by a startup company.
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Singer, M., Martin, L., Vargaftig, B. et al. A MARCKS-related peptide blocks mucus hypersecretion in a mouse model of asthma. Nat Med 10, 193–196 (2004). https://doi.org/10.1038/nm983
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DOI: https://doi.org/10.1038/nm983
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