Molecular principles for heparin oligosaccharide-based inhibition of neutrophil elastase in cystic fibrosis.

Abstract

Cystic fibrosis (CF) is a multifactorial disease in which dysfunction of protease-antiprotease balance plays a key role. The current CF therapy relies on dornase α, hypertonic saline, and antibiotics and does not address the high neutrophil elastase (NE) activity observed in the lung and sputum of CF patients. Our hypothesis is that variants of heparin, which potently inhibit NE but are not anticoagulant, would help restore the protease-antiprotease balance in CF. To realize this concept, we studied molecular principles governing the effectiveness of different heparins, especially 2-O,3-O-desulfated heparin (ODSH), in the presence of sputum components and therapeutic agents. Using sputa from CF patients and an NE activity assay, we found that heparins are ineffective if used in the absence of dornase. This is true even when mucolytics, such as DTT or N-acetylcysteine, were used. Computational modeling suggested that ODSH and DNA compete for binding to an overlapping allosteric site on NE, which reduces the anti-NE potential of ODSH. NE inhibition of both DNA and ODSH is chain length-dependent, but ODSH chains exhibit higher potency per unit residue length. Likewise, ODSH chains exhibit higher NE inhibition potential compared with DNA chains in the presence of saline. These studies suggest fundamental differences in DNA and ODSH recognition and inhibition of NE despite engaging overlapping sites and offer unique insights into molecular principles that could be used in developing antiprotease agents in the presence of current treatments, such as dornase and hypertonic saline.

• Authors: Apparao B Kummarapurugu, Daniel K Afosah, Nehru Viji Sankaranarayanan, Rahaman Navaz Gangji, Shuo Zheng, Thomas Kennedy, Bruce K Rubin, Judith A Voynow, Umesh R Desai
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 293 Issue 32 Pg. 12480-12490 (08 10 2018) ISSN: 1083-351X [Electronic] United States
• PMID: 29903912 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: © 2018 Kummarapurugu et al.

Chemical References

• Oligosaccharides
• Protease Inhibitors
• heparin, O-desulfated
• Heparin
• Leukocyte Elastase

Topics

• Computer Simulation
• Cystic Fibrosis (physiopathology)
• Heparin (analogs & derivatives, pharmacology)
• Humans
• Leukocyte Elastase (antagonists & inhibitors, metabolism)
• Oligosaccharides (pharmacology)
• Protease Inhibitors (pharmacology)
• Sputum (enzymology)