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Clinical Journal of the American... Apr 2014
Topics: Acute Kidney Injury; Acute-Phase Proteins; Clinical Enzyme Tests; Female; Humans; Leukocyte Elastase; Lipocalin-2; Lipocalins; Male; Matrix Metalloproteinase 8; Proto-Oncogene Proteins; Sepsis
PubMed: 24677556
DOI: 10.2215/CJN.01520214 -
MEDICC Review Oct 2022In inflammatory respiratory diseases, the imbalance between proteases and endogenous protease inhibitors leads to an exacerbated activity of human neutrophil elastase (a...
INTRODUCTION
In inflammatory respiratory diseases, the imbalance between proteases and endogenous protease inhibitors leads to an exacerbated activity of human neutrophil elastase (a protease that destroys the extracellular matrix and stimulates proinflammatory cytokine release). Elastase is considered a target in the search for therapeutic treatments for inflammatory respiratory diseases. Pulmonary surfactant is a promising product for this purpose, because in addition to its biophysical function, it has anti-inflammatory properties.
OBJECTIVE
Evaluate effect of the Cuban porcine pulmonary surfactant (Surfacen), the rCmPI-II elastase inhibitor, and the Surfacen/rCmPI-II combination on activated neutrophil elastase activity in vitro, and determine if Surfacen's interface property changes in the presence of the inhibitor.
METHODS
The anti-elastase effect of Surfacen, rCmPI-II and the Surfacen/rCmPI-II combination was evaluated in an in vitro model of activated neutrophils, previously purified from the blood of healthy subjects. The cells were stimulated with LPS/fMLP and were incubated with different concentrations of Surfacen, rCmPI-II and the Surfacen/rCmPI-II combination. Elastase activity was measured. The interface property was determined on a Langmuir surface balance. The new index, called the abdominal adipose deposit index, was obtained by multiplying the subcutaneous fat thickness by visceral fat thickness, both measured by ultrasound. A cutoff point was established that facilitated discernment of an unhealthy phenotype: normal weight but metabolically obese, a cardiometabolic risk factor.
RESULTS
Surfacen at 10 mg/mL inhibited 71% of stimulated neutrophil elastase activity. rCmPI-II at 0.1 μM reduced 20% of elastase activity; at 200 μM-the maximum concentration evaluated-inhibition was 68%. Both products had a dose-dependent effect. The Surfacen/inhibitor combination (0.5 mg/mL/80 µM) did not affect the surfactant interface property or the inhibitory activity of rCmPI-II against human neutrophil elastase.
CONCLUSIONS
Surfacen and the rCmPI-II inhibitor have an anti-elastase effect on an activated neutrophil model. rCmPI-II does not affect Surfacen's interface property and, therefore, both can be evaluated for combined use in treating inflammatory lung diseases.
Topics: Animals; Humans; Antiviral Agents; Leukocyte Elastase; Neutrophils; Protease Inhibitors; Pulmonary Surfactants; Swine
PubMed: 36417334
DOI: 10.37757/MR2022.V24.N3-4.7 -
Expert Opinion on Therapeutic Targets Feb 2008Cystic fibrosis (CF) is a lethal hereditary disease characterised by neutrophil-dominated lung inflammation. These abundant neutrophils produce neutrophil elastase (NE),... (Review)
Review
BACKGROUND
Cystic fibrosis (CF) is a lethal hereditary disease characterised by neutrophil-dominated lung inflammation. These abundant neutrophils produce neutrophil elastase (NE), a destructive serine protease that has direct actions on extracellular matrix proteins and has a role in the host response to inflammation and infection.
OBJECTIVE
This review examines the prospect of developing novel therapies for CF by targeting NE. The authors explore the functions of NE and of naturally-occurring and synthetic NE inhibitors.
METHODS
A literature search was conducted exploring the functions of NE and inhibitors of NE; naturally occurring and synthetic.
CONCLUSIONS
Targeting NE in CF offers therapeutic potential, but optimal inhibitors that can be delivered safely and effectively to the lung are still under development.
Topics: Cystic Fibrosis; Enzyme Inhibitors; Humans; Leukocyte Elastase
PubMed: 18208364
DOI: 10.1517/14728222.12.2.145 -
Bioorganic & Medicinal Chemistry Dec 2020We describe the total synthesis of tutuilamide A, a potent porcine pancreatic elastase (PPE) inhibitor and a representative member of the 3-amino-6-hydroxy-2-piperidone...
Ahp-Cyclodepsipeptides as tunable inhibitors of human neutrophil elastase and kallikrein 7: Total synthesis of tutuilamide A, serine protease selectivity profile and comparison with lyngbyastatin 7.
We describe the total synthesis of tutuilamide A, a potent porcine pancreatic elastase (PPE) inhibitor and a representative member of the 3-amino-6-hydroxy-2-piperidone (Ahp) cyclodepsipeptide family, isolated from marine cyanobacteria. The Ahp unit serves as a pharmacophore and the adjacent 2-amino-2-butenoic acid (Abu) is a main driver of the selectivity among serine proteases. We adapted our previous convergent strategy to generate the macrocycle, common with lyngbyastatin 7 and related elastase inhibitors, and then appended the tutuilamide A-specific side chain bearing a vinyl chloride. Tutuilamide A and lyngbyastatin 7 were evaluated side by side for the inhibition of the disease-relevant human neutrophil elastase (HNE). Tutuilamide A and lyngbyastatin 7 were approximately equipotent against HNE, while tutuilamide A was previously shown to be more active against PPE compared with lyngbyastatin 7, further demonstrating that the side chain provides opportunities to not only modulate potency but also selectivity among proteases of the same function from different organisms. Profiling of tutuilamide A against mainly human serine proteases revealed high selectivity for HNE (IC 0.73 nM) and pleiotropic activity against kallikrein 7 (KLK7, IC 5.0 nM), without affecting other kallikreins, similarly to lyngbyastatin 7 (IC 0.85 nM for HNE and 3.1 nM for KLK7). A comprehensive molecular docking study for elastases and KLK7 afforded deeper insight into the intricate differences between inhibitor interactions with HNE and PPE, accounting for the differential activities for both compounds. The synthesis and molecular studies serve as a proof-of-concept that the macrocyclic scaffold can be diversified to fine-tune the activity of serine protease inhibitors.
Topics: Binding Sites; Depsipeptides; Humans; Kallikreins; Kinetics; Leukocyte Elastase; Molecular Docking Simulation; Peptide Hydrolases; Serine Proteinase Inhibitors
PubMed: 33002682
DOI: 10.1016/j.bmc.2020.115756 -
Frontiers in Immunology 2020Neutrophil extracellular traps (NETs) are a defense mechanism in which neutrophils cast a net-like structure in response to microbial infection. NETs consist of...
BACKGROUND
Neutrophil extracellular traps (NETs) are a defense mechanism in which neutrophils cast a net-like structure in response to microbial infection. NETs consist of decondensed chromatin and about 30 enzymes and peptides. Some components, such as neutrophil elastase (NE) and myeloperoxidase (MPO), present antimicrobial but also cytotoxic properties, leading to tissue injury. Many inflammatory diseases are associated with NETs, and their final role has not been identified. Pulmonary surfactant is known to have immunoregulatory abilities that alter the function of adaptive and innate immune cells. The aim of this study was to investigate the hypothesis that natural surfactant preparations inhibit the formation of NETs.
METHODS
The effect of two natural surfactants (Alveofact and Curosurf) on spontaneous and phorbol-12-myristate-13-acetate-induced NET formation by neutrophils isolated by magnetic cell sorting from healthy individuals was examined. NETs were quantitatively detected by absorption and fluorometric-based assays for the NET-specific proteins (NE, MPO) and cell-free DNA. Immunofluorescence microscopy images were used for visualization.
RESULTS
Both surfactant preparations exerted a dose-dependent inhibitory effect on NET formation. Samples treated with higher concentrations and with 30 min pre-incubation prior to stimulation with phorbol-12-myristate-13-acetate had significantly lower levels of NET-specific proteins and cell-free DNA compared to untreated samples. Immunofluorescence microscopy confirmed these findings.
CONCLUSIONS
The described dose-dependent modulation of NET formation ex vivo suggests an interaction between exogenous surfactant supplementation and neutrophil granulocytes. The immunoregulatory effects of surfactant preparations should be considered for further examination of inflammatory diseases.
Topics: Biological Products; Cells, Cultured; Dose-Response Relationship, Drug; Extracellular Traps; Granulocytes; Humans; Immunomodulation; Leukocyte Elastase; Neutrophil Activation; Neutrophils; Phospholipids; Pulmonary Surfactants; Tetradecanoylphorbol Acetate
PubMed: 33574811
DOI: 10.3389/fimmu.2020.582895 -
The FEBS Journal Nov 2012This study aimed to investigate the susceptibility of intact fibrillar human elastin to human leukocyte elastase and cathepsin G. Elastin is a vital protein of the...
This study aimed to investigate the susceptibility of intact fibrillar human elastin to human leukocyte elastase and cathepsin G. Elastin is a vital protein of the extracellular matrix of vertebrates, and provides exceptional properties including elasticity and tensile strength to many tissues and organs, including the aorta, lung, cartilage, elastic ligaments and skin, and is thus critical for their long-term function. Mature elastin is an insoluble and extremely durable protein that undergoes very little turnover, but sustained exposure to proteases may lead to irreversible and severe damage, and thus to functional loss of the elastic fiber network. Hence, it is a key issue to understand which enzymes actually initiate elastolysis under certain pathological conditions or during intrinsic aging. In this paper, we provide a complete workflow for isolation of pure and intact elastin from very small tissue samples to test enzymes for their elastolytic potential. This workflow was applied to skin samples from variously aged individuals, and it was found that strong differences exist in the degradability of the elastins investigated. In summary, human leukocyte elastase was unable to degrade intact elastin fibers but hydrolyzed elastin derived from the skin of old people. However, cathepsin G cleaved all elastin samples, even those derived from younger individuals. These results indicate that human leukocyte elastase is not a driving force for elastolysis, but may nevertheless promote further breakdown of elastic fibers after the action of other enzymes such as cathepsin G.
Topics: Adult; Aged, 80 and over; Cathepsin G; Child; Elastin; Female; Humans; Leukocyte Elastase; Microscopy, Electron, Scanning; Skin; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
PubMed: 23006486
DOI: 10.1111/febs.12012 -
Bioscience Trends Jan 2020Significant advances in our understanding of neutrophil biology were made in the past several years. A newly discovered mechanism was discovered, the formation of... (Review)
Review
Significant advances in our understanding of neutrophil biology were made in the past several years. A newly discovered mechanism was discovered, the formation of neutrophils extracellular traps (NETs). The structure of NETs is composed of the DNA strand and neutrophil granule proteins. NETs were found to have an association with tumor progression. This review highlights the latest knowledge about the controversial effect on tumors of NETs. Pro-tumor and anti-tumor effects are described respectively. The probable mechanisms of the anti-tumor effect are related to its direct killing of cancer cells or stimulation of the immune system to fight against the tumor. The pro-tumor effect has a correlation with matrix metalloproteinase 9 (MMP-9), cathepsin G, and neutrophil elastase (NE). Moreover, the structure of the NETs makes it able to catch the circulating tumor cells, which could lead to metastasis. This review summarizes our knowledge about the proven roles of NETs in the progression of cancer with particular focus on the components of the NETs, and considers NETs as a potential target for cancer therapy.
Topics: Animals; Cathepsin G; Extracellular Traps; Humans; Leukocyte Elastase; Matrix Metalloproteinase 9; Neutrophils
PubMed: 31866615
DOI: 10.5582/bst.2019.01326 -
American Journal of Physiology. Lung... Mar 2022Muco-obstructive lung diseases are characterized by airway obstruction and hyperinflation, which can be quantified by imaging. Our aim was to evaluate µCT for...
Muco-obstructive lung diseases are characterized by airway obstruction and hyperinflation, which can be quantified by imaging. Our aim was to evaluate µCT for longitudinal quantification of muco-obstructive lung disease in β-epithelial Na channel overexpressing (-TG) mice and of the effects of neutrophil elastase (NE) knockout on its progression. Lungs from wild-type (WT), NE, -TG, and -TG/NE mice were scanned with 9-µm resolution at 0, 5, 14, and 60 days of age, and airway and parenchymal disease was quantified. Mucus adhesion lesions (MAL) were persistently increased in -TG compared with WT mice from 0 days (20.25 ± 6.50 vs. 9.60 ± 2.07, < 0.05), and this effect was attenuated in -TG/NE mice (5.33 ± 3.67, < 0.001). Airway wall area percentage (WA%) was increased in -TG mice compared with WT from 14 days onward (59.2 ± 6.3% vs. 49.8 ± 9.0%, < 0.001) but was similar in -TG/NE compared with WT at 60 days (46.4 ± 9.2% vs. 45.4 ± 11.5%, = 0.97). Air proportion (Air%) and mean linear intercept () were persistently increased in -TG compared with WT from 5 days on (53.9 ± 4.5% vs. 30.0 ± 5.5% and 78.82 ± 8.44 µm vs. 65.66 ± 4.15 µm, respectively, < 0.001), whereas in -TG/NE, Air% and were similar to WT from birth (27.7 ± 5.5% vs. 27.2 ± 5.9%, = 0.92 and 61.48 ± 9.20 µm vs. 61.70 ± 6.73 µm, = 0.93, respectively). Our results suggest that µCT is sensitive to detect the onset and progression of muco-obstructive lung disease and effects of genetic deletion of NE on morphology of airways and lung parenchyma in -TG mice, and that it may serve as a sensitive endpoint for preclinical studies of novel therapeutic interventions for muco-obstructive lung diseases.
Topics: Animals; Disease Models, Animal; Epithelial Sodium Channels; Leukocyte Elastase; Lung; Lung Diseases, Obstructive; Mice; Mice, Knockout; Mice, Transgenic
PubMed: 35080183
DOI: 10.1152/ajplung.00341.2021 -
Contrast Media & Molecular Imaging 2019The last few decades of protease research has confirmed that a number of important biological processes are strictly dependent on proteolysis. Neutrophil elastase (NE)... (Review)
Review
The last few decades of protease research has confirmed that a number of important biological processes are strictly dependent on proteolysis. Neutrophil elastase (NE) is a critical protease in immune response and host defense mechanisms in both physiological and disease-associated conditions. Particularly, NE has been identified as a promising biomarker for early diagnosis of lung inflammation. Recent studies have shown an increasing interest in developing methods for NE activity imaging both in vitro and in vivo. Unlike anatomical imaging modalities, functional molecular imaging, including enzymatic activities, enables disease detection at a very early stage and thus constitutes a much more accurate approach. When combined with advanced imaging technologies, opportunities arise for measuring imbalanced proteolytic activities with unprecedented details. Such technologies consist in building the highest resolved and sensitive instruments as well as the most specific probes based either on peptide substrates or on covalent inhibitors. This review outlines strengths and weaknesses of these technologies and discuss their applications to investigate NE activity as biomarker of pulmonary inflammatory diseases by imaging.
Topics: Animals; Asymptomatic Diseases; Biomarkers; Biopolymers; Catalytic Domain; Chromogenic Compounds; Cytoplasmic Granules; Early Diagnosis; Enzyme-Linked Immunosorbent Assay; Fluorescence Resonance Energy Transfer; Fluorescent Dyes; Humans; Imaging, Three-Dimensional; Leukocyte Elastase; Magnetic Resonance Imaging; Molecular Imaging; Neutrophils; Oligopeptides; Optical Imaging; Pneumonia; Positron-Emission Tomography; Proteolysis; Substrate Specificity
PubMed: 31281234
DOI: 10.1155/2019/7417192 -
Proceedings of the National Academy of... Sep 2013The rationale of α1-antitrypsin (AAT) augmentation therapy to treat progressive emphysema in AAT-deficient patients is based on inhibition of neutrophil elastase;...
The rationale of α1-antitrypsin (AAT) augmentation therapy to treat progressive emphysema in AAT-deficient patients is based on inhibition of neutrophil elastase; however, the benefit of this treatment remains unclear. Here we show that clinical grade AAT (with elastase inhibitory activity) and a recombinant form of AAT (rAAT) without anti-elastase activity reduces lung inflammatory responses to LPS in elastase-deficient mice. WT and elastase-deficient mice treated with either native AAT or rAAT exhibited significant reductions in infiltrating neutrophils (23% and 68%), lavage fluid levels of TNF-α (70% and 80%), and the neutrophil chemokine KC (CXCL1) (64% and 90%), respectively. Lung parenchyma TNF-α, DNA damage-inducible transcript 3 and X-box binding protein-1 mRNA levels were reduced in both mouse strains treated with AAT; significantly lower levels of these genes, as well as IL-1β gene expression, were observed in lungs of AAT-deficient patients treated with AAT therapy compared with untreated patients. In vitro, LPS-induced cytokines from WT and elastase-deficient mouse neutrophils, as well as neutrophils of healthy humans, were similarly reduced by AAT or rAAT; human neutrophils adhering to endothelial cells were decreased by 60-80% (P < 0.001) with either AAT or rAAT. In mouse pancreatic islet macrophages, LPS-induced surface expression of MHC II, Toll-like receptor-2 and -4 were markedly lower (80%, P < 0.001) when exposed to either AAT or rAAT. Consistently, in vivo and in vitro, rAAT reduced inflammatory responses at concentrations 40- to 100-fold lower than native plasma-derived AAT. These data provide evidence that the anti-inflammatory and immunomodulatory properties of AAT can be independent of elastase inhibition.
Topics: Acute Lung Injury; Adult; Animals; Anti-Inflammatory Agents; Cytokines; Disease Models, Animal; Female; Gene Expression; Humans; Immunologic Factors; Leukocyte Elastase; Lipopolysaccharides; Lung; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Middle Aged; Neutrophils; Pulmonary Emphysema; Recombinant Proteins; alpha 1-Antitrypsin; alpha 1-Antitrypsin Deficiency
PubMed: 23975926
DOI: 10.1073/pnas.1309648110