Elastase inhibitors as potential therapies for ELANE-associated neutropenia

Abstract

Mutations in , the gene for neutrophil elastase (NE), a protease expressed early in neutrophil development, are the most frequent cause of cyclic (CyN) and severe congenital neutropenia (SCN). We hypothesized that inhibitors of NE, acting either by directly inhibiting enzymatic activity or as chaperones for the mutant protein, might be effective as therapy for CyN and SCN. We investigated β-lactam-based inhibitors of human NE (Merck Research Laboratories, Kenilworth, NJ, USA), focusing on 1 inhibitor called MK0339, a potent, orally absorbed agent that had been tested in clinical trials and shown to have a favorable safety profile. Because fresh, primary bone marrow cells are rarely available in sufficient quantities for research studies, we used 3 cellular models: patient-derived, induced pluripotent stem cells (iPSCs); HL60 cells transiently expressing mutant NE; and HL60 cells with regulated expression of the mutant enzyme. In all 3 models, the cells expressing the mutant enzyme had reduced survival as measured with annexin V and FACS. Coincubation with the inhibitors, particularly MK0339, promoted cell survival and increased formation of mature neutrophils. These studies suggest that cell-permeable inhibitors of neutrophil elastase show promise as novel therapies for-associated neutropenia.

Differentially Expressed Genes in Osteomyelitis Induced by Staphylococcus aureus Infection

“…Module 1 was an interesting module associated with specific functions, including ELANE, MPO, CAMP, CTSG, etc. Mutations in ELANE often cause cyclic and severe congenital neutropenia (Makaryan et al, 2017 ). MPO has been recently demonstrated to be associated with NETs production (Granger et al, 2017 ; Kenny et al, 2017 ).…”

Neutrophil Elastase Defects in Congenital Neutropenia

“…However, there are studies that support alternative scenarios. In a cellular model of HL-60, human promyeloblast cell line transfected with mutant NE, none of the studied UPR markers, neither BiP/GRP78 nor ATF6 were expressed ( 83 ). Likewise, in a similar in vitro model of granulopoiesis was impaired in ELANE -mutated cell line, albeit the UPR was not induced ( 51 ).…”

Neutrophil Elastase Inhibitors and Chronic Kidney Disease

“…The design of small-molecule activity-based probes by click chemistry approach on 4-oxo-β-lactams was also reported as an efficient tool for the development of new potent NE inhibitors; these probes showed adequate fluorescence properties, and neutrophil internalization, suitable for detection of NE, making them a promise as diagnostic tools 85 . MK0339, a potent human β-lactam-based inhibitor, was tested in clinical trials, stimulating cell survival and increasing the production of mature neutrophils; data suggested MK0339 as a potential agent to be used in ELANE -associated neutropenia 86 . Derivatives of O3-acyl kojic acid, selected by virtual screening, are also under study as they presented good selectivity for NE and low cytotoxicity 87 .…”