Identification and characterization of aptameric inhibitors of human neutrophil elastase

Stanisław Malicki; Mirosław Książek; Alicja Sochaj Gregorczyk; Marta Kamińska; Anna Golda; Barbara Chruścicka; Danuta Mizgalska; Jan Potempa; Hans-Peter Marti; Joanna Kozieł; Maciej Wieczorek; Jerzy Pieczykolan; Piotr Mydel; Grzegorz Dubin

doi:10.1016/j.jbc.2023.104889

Identification and characterization of aptameric inhibitors of human neutrophil elastase

J Biol Chem. 2023 Aug;299(8):104889. doi: 10.1016/j.jbc.2023.104889. Epub 2023 Jun 5.

Affiliations

¹ Laboratory of Proteolysis and Post-translational Modification of Proteins, Malopolska Centre of Biotechnology, Jagiellonian University, Krakow, Poland; Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland. Electronic address: stanislaw.malicki@uj.edu.pl.
² Laboratory of Proteolysis and Post-translational Modification of Proteins, Malopolska Centre of Biotechnology, Jagiellonian University, Krakow, Poland; Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.
³ Broegelmann Research Laboratory, University of Bergen, Bergen, Norway.
⁴ Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.
⁵ Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland; Department of Oral Immunity and Infectious Diseases, University of Louisville School of Dentistry, Louisville, Kentucky, USA.
⁶ Department of Clinical Medicine, University of Bergen, Bergen, Norway.
⁷ Innovative Drugs R&D Department, Celon Pharma Inc, Lomianki, Poland.
⁸ Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland; Broegelmann Research Laboratory, University of Bergen, Bergen, Norway.
⁹ Protein Crystallography Research, Group Malopolska Centre of Biotechnology, Jagiellonian University, Krakow, Poland.

Abstract

Human neutrophil elastase (HNE) plays a pivotal role in innate immunity, inflammation, and tissue remodeling. Aberrant proteolytic activity of HNE contributes to organ destruction in various chronic inflammatory diseases including emphysema, asthma, and cystic fibrosis. Therefore, elastase inhibitors could alleviate the progression of these disorders. Here, we used the systematic evolution of ligands by exponential enrichment to develop ssDNA aptamers that specifically target HNE. We determined the specificity of the designed inhibitors and their inhibitory efficacy against HNE using biochemical and in vitro methods, including an assay of neutrophil activity. Our aptamers inhibit the elastinolytic activity of HNE with nanomolar potency and are highly specific for HNE and do not target other tested human proteases. As such, this study provides lead compounds suitable for the evaluation of their tissue-protective potential in animal models.

Keywords: aptamer; elastase; human neutrophil elastase; neutrophil; pulmonary disease.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Aptamers, Nucleotide* / chemical synthesis
Aptamers, Nucleotide* / pharmacology
Aptamers, Nucleotide* / therapeutic use
Cells, Cultured
Cystic Fibrosis / drug therapy
Emphysema / drug therapy
Enzyme Activation / drug effects
Humans
Leukocyte Elastase* / antagonists & inhibitors
Neutrophils / drug effects
Proteolysis / drug effects
Sensitivity and Specificity
Serine Proteinase Inhibitors* / chemical synthesis
Serine Proteinase Inhibitors* / pharmacology
Serine Proteinase Inhibitors* / therapeutic use

Substances

Leukocyte Elastase
Serine Proteinase Inhibitors
Aptamers, Nucleotide