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Serpin regulation of leukocyte proteases in cellular homeostasis and inflammation

English title Serpin regulation of leukocyte proteases in cellular homeostasis and inflammation
Applicant Benarafa Charaf
Number 149790
Funding scheme Project funding (Div. I-III)
Research institution Theodor Kocher Institut Universität Bern
Institution of higher education University of Berne - BE
Main discipline Immunology, Immunopathology
Start/End 01.12.2013 - 31.05.2017
Approved amount 428'600.00
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All Disciplines (3)

Discipline
Immunology, Immunopathology
Biochemistry
Cellular Biology, Cytology

Keywords (6)

Protease; Neutrophils; Lung; Inflammation; Serpins; Apoptosis

Lay Summary (French)

Lead
Le but de notre recherche est d’identifier de nouveaux mécanismes cellulaires et moléculaires liés à la régulation des protéases par les serpines intracellulaires des cellules du système immunitaire dans les processus de mort cellulaire et dans les maladies inflammatoires.
Lay summary

Les protéases stockées dans les granules des cellules du système immunitaire sont des enzymes qui contribuent à la fois aux fonctions protectrices et destructrices de l’inflammation. Ces protéases tuent directement les agents pathogènes, les cellules cancéreuses ou celles infectées par des virus, et inactivent les toxines bactériennes. Cependant, un excès de protéases contribue également à la destruction des tissus et amplifie la réaction inflammatoire au détriment de l’hôte. La mort des cellules inflammatoires comme les neutrophiles contribue dans une large mesure à la libération d’un excès de protéases et alimente un cercle vicieux pro-inflammatoire. De nombreuses études ont porté sur la régulation des protéases par des inhibiteurs sécrétés tels que l’antitrypsine du plasma. Nos recherches récentes ont permis d’identifier un rôle protecteur prépondérant pour l’inhibiteur intracellulaire serpinB1 qui bloque l’activité des protéases des neutrophiles dans les infections pulmonaires et qui régule la survie des neutrophiles in vivo. Nous proposons d'élucider le rôle d’un groupe de serpines intracellulaires (dont serpinB1) dans les mécanismes de mort cellulaire et de l’inflammation et d'identifier l'importance relative des différentes protéases et serpines dans ces processus. Ces études seront utiles pour mieux cibler les protéases impliquées dans la mort cellulaire et l'inflammation dans les pathologies infectieuses et non infectieuses.

Direct link to Lay Summary Last update: 08.10.2013

Responsible applicant and co-applicants

Employees

Publications

Publication
Role of granule proteases in the life and death of neutrophils.
Benarafa Charaf, Simon Hans-Uwe (2017), Role of granule proteases in the life and death of neutrophils., in Biochemical and biophysical research communications, 482, 473-481.
Adamts18 deletion results in distinct developmental defects and provides a model for congenital disorders of lens, lung, and female reproductive tract development.
Ataca Dalya, Caikovski Marian, Piersigilli Alessandra, Moulin Alexandre, Benarafa Charaf, Earp Sarah E, Guri Yakir, Kostic Corinne, Arsenivic Yvan, Soininen Raija, Apte Suneel S, Brisken Cathrin (2016), Adamts18 deletion results in distinct developmental defects and provides a model for congenital disorders of lens, lung, and female reproductive tract development., in Biology open, 5, 1585-1594.
Distinct gene expression patterns correlate with developmental and functional traits of iNKT subsets.
Georgiev Hristo, Ravens Inga, Benarafa Charaf, Förster Reinhold, Bernhardt Günter (2016), Distinct gene expression patterns correlate with developmental and functional traits of iNKT subsets., in Nature communications, 7, 13116-13116.
Exogenous cathepsin G upregulates cell surface MHC class I molecules on immune and glioblastoma cells.
Giese Madleen, Turiello Nadine, Molenda Nicole, Palesch David, Meid Annika, Schroeder Roman, Basilico Paola, Benarafa Charaf, Halatsch Marc-Eric, Zimecki Michal, Westhoff Mike-Andrew, Wirtz Christian Rainer, Burster Timo (2016), Exogenous cathepsin G upregulates cell surface MHC class I molecules on immune and glioblastoma cells., in Oncotarget, 7, 74602-74611.
Increased Myeloid Cell Production and Lung Bacterial Clearance in Mice Exposed to Cigarette Smoke.
Basilico Paola, Cremona Tiziana Patrizia, Oevermann Anna, Piersigilli Alessandra, Benarafa Charaf (2016), Increased Myeloid Cell Production and Lung Bacterial Clearance in Mice Exposed to Cigarette Smoke., in American journal of respiratory cell and molecular biology, 54, 424-435.
Myeloid conditional deletion and transgenic models reveal a threshold for the neutrophil survival factor Serpinb1.
Burgener Sabrina S, Baumann Mathias, Basilico Paola, Remold-O&apos, Donnell Eileen, Touw Ivo P, Benarafa Charaf (2016), Myeloid conditional deletion and transgenic models reveal a threshold for the neutrophil survival factor Serpinb1., in Biological Chemistry, 397, 897-905.
RhoH is a negative regulator of eosinophilopoiesis.
Stoeckle Christina, Geering Barbara, Yousefi Shida, Rozman Sasa, Andina Nicola, Benarafa Charaf, Simon Hans-Uwe (2016), RhoH is a negative regulator of eosinophilopoiesis., in Cell death and differentiation, 23, 1961-1972.
Small angle x-ray scattering with edge-illumination.
Modregger Peter, Cremona Tiziana Patrizia, Benarafa Charaf, Schittny Johannes C, Olivo Alessandro, Endrizzi Marco (2016), Small angle x-ray scattering with edge-illumination., in Scientific reports, 6, 30940-30940.
CXCL14 displays antimicrobial activity against respiratory tract bacteria and contributes to clearance of Streptococcus pneumoniae pulmonary infection.
Dai Chen, Basilico Paola, Cremona Tiziana Patrizia, Collins Paul, Moser Bernhard, Benarafa Charaf, Wolf Marlene (2015), CXCL14 displays antimicrobial activity against respiratory tract bacteria and contributes to clearance of Streptococcus pneumoniae pulmonary infection., in The Journal of Immunology, 194, 5980-5989.
CXCL14: the Swiss army knife chemokine.
Benarafa Charaf, Wolf Marlene (2015), CXCL14: the Swiss army knife chemokine., in Oncotarget, 6, 34065-34066.
Neutrophils: Between Host Defence, Immune Modulation, and Tissue Injury.
Kruger Philipp, Saffarzadeh Mona, Weber Alexander N R, Rieber Nikolaus, Radsak Markus, von Bernuth Horst, Benarafa Charaf, Roos Dirk, Skokowa Julia, Hartl Dominik (2015), Neutrophils: Between Host Defence, Immune Modulation, and Tissue Injury., in PLoS pathogens, 11, 1004651-1004651.
Regulation of Neutrophil Serine Proteases by Intracellular Serpins
Benarafa Charaf (2015), Regulation of Neutrophil Serine Proteases by Intracellular Serpins, in Geiger Margarethe (ed.), Springer, Switzerland, 329-329.
The generation of neutrophils in the bone marrow is controlled by autophagy.
Rožman S, Yousefi S, Oberson K, Kaufmann T, Benarafa Charaf, Simon H U (2015), The generation of neutrophils in the bone marrow is controlled by autophagy., in Cell death and differentiation, 22, 445-456.
Tumor-induced inflammation alters neutrophil phenotype and disease progression.
Benarafa Charaf (2015), Tumor-induced inflammation alters neutrophil phenotype and disease progression., in Breast cancer research : BCR, 17, 135-135.
DAPK2 positively regulates motility of neutrophils and eosinophils in response to intermediary chemoattractants.
Geering Barbara, Stoeckle Christina, Rozman Sasa, Oberson Kevin, Benarafa Charaf, Simon Hans-Uwe (2014), DAPK2 positively regulates motility of neutrophils and eosinophils in response to intermediary chemoattractants., in Journal of leukocyte biology, 95, 293-303.

Collaboration

Group / person Country
Types of collaboration
Prof. Bird, Monash University, Australia Australia (Oceania)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Prof. Bernhardt, Hannover Medical School, Hannover Germany (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Prof. Simon, Institute for Pharmacology, University of Bern Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Prof. Hartl, University of Tübingen Germany (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Prof Brisken, EPFL Switzerland (Europe)
- Publication
Prof. Pham, University of Washington, St Louis, MO United States of America (North America)
- in-depth/constructive exchanges on approaches, methods or results
- Publication

Scientific events

Active participation

Title Type of contribution Title of article or contribution Date Place Persons involved
VIA Congress 2017 - Deutsche Gesellschaft für Immunologie (DGfI) Talk given at a conference Intracellular serpins regulate multiple programmed cell death pathways in leukocytes 27.04.2017 Bern, Switzerland Benarafa Charaf;
Annual Symposium - Graduate School for Cellular and Biomedical Sciences, University of Bern Individual talk Suicide in paradise: Neutrophil survival depends on Serpinb1a and Serpinb6a inhibition of granule proteases 02.02.2017 Bern, Switzerland Burgener Sabrina Sofia;
Seminar Series, German Mouse Clinic, Helmholtz Centrum Münich Individual talk Phenotyping of mice lacking homologs of serpinb1 serpinb6 and serpinb9 22.12.2016 Münich, Germany Benarafa Charaf;
Invited talk - Institute for virology and immunology Individual talk Functions of intracellular serpins in immunity and inflammation 19.12.2016 Mittelhaüsern, Switzerland Benarafa Charaf;
Cutting Edge Topics: Seminars in Immunology and Infection Biology Individual talk Regulation of myeloid cell homeostasis by intracellular serpins 20.09.2016 University of Zürich, Switzerland Benarafa Charaf;
49th Meeting of the Society for Leukocyte Biology & "Neutrophil 2016" Talk given at a conference Neutrophil survival depends on Serpinb1a and Serpinb6a inhibition of granule proteases 15.09.2016 Verona, Italy Benarafa Charaf; Burgener Sabrina Sofia;
15th International Summer School, Bern Immunology Club Talk given at a conference Functions of intracellular serpins in serine protease-mediated death 07.08.2016 Bönigen, Switzerland Benarafa Charaf;
Invited seminar - Unité INSERM 1148-LVTS, CHU Xavier Bichat Individual talk Functions of intracellular serpins in neutrophil serine protease-mediated death 23.10.2015 CHU Xavier Bichat, Paris, France Benarafa Charaf;
23rd Conference of the European Cell Death Organisation (ECDO) Poster Additive effect of serpinB1 and serpinB6 in preventing cell death in neutrophils 07.10.2015 Geneva, Switzerland Burgener Sabrina Sofia;
9th General Meeting of the International Proteolysis Society, Penang, Malaysia Talk given at a conference Functions of intracellular serpins in neutrophil serine protease-mediated death 04.10.2015 Penang, Malaysia Benarafa Charaf;
14th International Summer School, Bern Immunology Club Talk given at a conference Everything is a question of dose – additive effect of serpinB1 and serpinB6 in preventing cell death in neutrophils 26.07.2015 Bönigen, Switzerland Burgener Sabrina Sofia;
14th International Summer School, Bern Immunology Club Talk given at a conference Cigarette smoke alters the clearance of bacterial pathogens in the respiratory tract 26.07.2015 Bönigen, Switzerland Benarafa Charaf;
1st Meeting of the Neutrophil Club, French Society for Immunology Talk given at a conference Granule serine proteases: the enemy within 10.04.2015 Paris, France Benarafa Charaf;
Seminar Series, Institute for Cell Biology, University of Bern Individual talk Intracellular serpins: safekeepers in cell death and inflammation 27.10.2014 Bern, Switzerland Benarafa Charaf;
8th Swiss Apoptosis Meeting Poster Cytoprotective function of serpinB1 and serpinB6 against neutrophil proteases in lung epithelial cells 10.09.2014 Bern, Switzerland Benarafa Charaf; Burgener Sabrina Sofia;
Annual Congress of the European Respiratory Society Talk given at a conference Increased myelopoiesis induced by cigarette smoke transiently improves kinetics of lung bacterial clearance in mice 06.09.2014 Münich, Germany Basilico Paola;
Annual Congress of the European Respiratory Society Poster Increased myelopoiesis induced by cigarette smoke transiently improved kinetics of lung bacterial clearance in mice 06.09.2014 Münich, Germany Benarafa Charaf; Basilico Paola;
13th International Summer School, Bern Immunology Club Talk given at a conference Neutrophil Granule Proteases and Serpins: Lessons from human diseases and animal models 28.07.2014 Stam am Rhein, Switzerland Benarafa Charaf;
Invited seminar - Dept. Pediatrics, University Children’s Hospital, Tübingen Individual talk Functions of serpinb1 in neutrophil homeostasis 08.07.2014 Tübingen, Germany Benarafa Charaf;
Seminar Series - University of North Carolina, Chapel Hill Individual talk Functions of serpinB1 in neutrophil homeostasis and pulmonary diseases 06.06.2014 Chapel Hill, North Carolina, United States of America Benarafa Charaf;
Neutrophil 2014 Poster Altered myelopoeisis and bacterial clearance in neutropenic serpinB1-/- mice following G-CSF therapy and cigarette smoke exposure 31.05.2014 Montreal, Canada Benarafa Charaf;
American Thoracic Society 2014 International Conference Poster Cigarette smoke exposure induces systemic inflammation with an acute effect on myelopoiesis and lung bacterial clearance 16.05.2014 San Diego, United States of America Basilico Paola;
48th Annual Scientific Meeting of the European Society for Clinical Investigation Poster Cigarette smoke exposure transiently increases myelopoiesis and bacterial clearance in wild-type and serpinB1-/- mice 30.04.2014 Utrecht, Netherlands Basilico Paola; Benarafa Charaf;
7th International Symposium on Serpin Biology, Structure and Function Talk given at a conference Systemic inflammation induced by cigarette smoke exposure transiently increases myelopoiesis and lung bacterial clearance in serpinB1-/- mice 29.03.2014 Leogang, Austria Basilico Paola;
7th International Symposium on Serpin Biology, Structure and Function Talk given at a conference Regulation of neutrophil survival by serpinB1 29.03.2014 Leogang, Austria Benarafa Charaf;
7th International Symposium on Serpin Biology, Structure and Function Talk given at a conference Cytoprotective regulation of neutrophil serine proteases by MNEI (serpinB1) and PI-6 (serpinB6) in lung epithelial cells 29.03.2014 Leogang, Austria Burgener Sabrina Sofia;
LS2 Meeting Poster Cytoprotective role of the intracellular serine protease inhibitors SerpinB1 and SerpinB6 in lung epithelial cells 04.02.2014 Lausanne, Switzerland Burgener Sabrina Sofia;
Meeting of the European Thoracic Surgery Group Talk given at a conference Proteases and serpins: at the intersection of lung inflammation, infection and cell death 17.01.2014 Bern, Switzerland Benarafa Charaf;


Communication with the public

Communication Title Media Place Year
Talks/events/exhibitions Génétique & Environnement: effets sur l’emphysème pulmonaire et la réponse aux infections Western Switzerland 2015

Awards

Title Year
Presidential Award of the Society for Leukocyte Biology http://leukocytebiology.org/Awards/SLB-Presidential-Award.aspx 2016

Associated projects

Number Title Start Funding scheme
127464 Role of serpinB1 in cellular homeostasis in the bone marrow and the lung 01.10.2009 Project funding (Div. I-III)
173137 Serpin regulation of leukocyte proteases in cellular homeostasis and inflammation 01.06.2017 Project funding (Div. I-III)
173137 Serpin regulation of leukocyte proteases in cellular homeostasis and inflammation 01.06.2017 Project funding (Div. I-III)

Abstract

Inflammatory responses are vital to preserve host integrity and survival following infection with pathogens. Genetic and environmental factors, however, contribute to modify, enhance or sustain these responses leading inflammatory diseases. Excessive acute amplification can lead to severe disease and lethality such as in acute respiratory distress syndrome and septic shock. Chronic inflammation is associated with sustained release of cytokines and enzymes leading to altered antimicrobial responses, reduced organ function and tissue destruction such as in cystic fibrosis (CF), asthma or chronic obstructive pulmonary disease (COPD) and emphysema. Serine proteases carried in neutrophil and mast cell granules are indiscriminate in their activity and have been shown to contribute to both protective and destructive inflammatory functions. These proteases directly kill pathogens and inactivate toxins but they also induce discrete proteolysis of host proteins with often detrimental consequences. Chronic release of these enzymes in the extracellular milieu, like in CF lung disease, COPD and arthritis, is thought to drive sustained tissue injury and inflammation. At these inflammatory sites, neutrophil death contributes to a large extent to excess protease release and fuels a cycle of that leads to increased neutrophil recruitment and tissue damage. Many studies have focused on neutrophil elastase, its targets and its regulation by plasma and secreted protease inhibitors such as a1-antitrypsin. We propose to investigate the role of the little studied, but increasingly recognized, family of intracellular (clade B) serpins in cellular homeostasis and inflammation.We have previously established that serpinB1, a potent intracellular inhibitor of neutrophil granule proteases (elastase, cathepsin G and proteinase-3), is required to prevent early neutrophil death both at inflammatory sites as well as in the bone marrow. Mice lacking serpinB1 have a severely reduced reserve of mature neutrophils in the bone marrow; they fail to clear Pseudomonas lung infection and have increased morbidity in influenza A virus infection. In these models, lack of serpinB1 is also associated with increased and sustained production of pro-inflammatory cytokines. We have recently identified that the cytoprotective role of serpinB1 depends on the inhibition of cathepsin G in a cell-intrinsic manner. Cathepsin G, but not neutrophil elastase, induced neutrophil death upon granule membrane permeabilization. In the first aim of this proposal, we will define the molecular and cellular signals that trigger the intracellular release of cathepsin G from granules leading to cell death in neutrophils in steady state and in inflammation. We will also investigate the function of serpinB1 and related serpins in mast cell homeostasis and granule-mediated death. In the second aim, we will investigate molecular mechanisms by which serpinB1 regulates neutrophil proteases in mouse models of inflammation. The proposed studies will generate novel findings on the fundamental cytoprotective functions of intracellular serpins and identify the relative importance of different granule proteases and serpins in amplifying and sustaining inflammation. These studies will be highly relevant for better targeting detrimental feedback loops involved in cell death and inflammation in infectious and non-infectious pathologies.
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