Neutrophil; Serpin; Mouse models; Cell death; Proteases; Immunity
Burgener Sabrina Sofia, Brügger Melanie, Leborgne Nathan Georges François, Sollberger Sophia, Basilico Paola, Kaufmann Thomas, Bird Phillip Ian, Benarafa Charaf (2021), Granule Leakage Induces Cell-Intrinsic, Granzyme B-Mediated Apoptosis in Mast Cells, in Frontiers in Cell and Developmental Biology
, 9, 1-12.
Germic Nina, Hosseini Aref, Stojkov Darko, Oberson Kevin, Claus Meike, Benarafa Charaf, Calzavarini Sara, Angelillo-Scherrer Anne, Arnold Isabelle C., Müller Anne, Riether Carsten, Yousefi Shida, Simon Hans-Uwe (2021), ATG5 promotes eosinopoiesis but inhibits eosinophil effector functions, in Blood
, 137(21), 2958-2969.
Hilty Markus, Wüthrich Tsering M., Godel Aurélie, Adelfio Roberto, Aebi Susanne, Burgener Sabrina S., Illgen-Wilcke Brunhilde, Benarafa Charaf (2020), Chronic cigarette smoke exposure and pneumococcal infection induce oropharyngeal microbiota dysbiosis and contribute to long-lasting lung damage in mice, in Microbial Genomics
, 6(12), 1-14.
Leborgne Nathan G. F., Taddeo Adriano, Freigang Stefan, Benarafa Charaf (2020), Serpinb1a Is Dispensable for the Development and Cytokine Response of Invariant Natural Killer T Cell Subsets, in Frontiers in Immunology
, 11, 1-9.
Ohayon Delphine, De Chiara Alessia, Dang Pham My-Chan, Thieblemont Nathalie, Chatfield Simon, Marzaioli Viviana, Burgener Sabrina Sofia, Mocek Julie, Candalh Céline, Pintard Coralie, Tacnet-Delorme Pascale, Renault Gilles, Lagoutte Isabelle, Favier Maryline, Walker Francine, Hurtado-Nedelec Margarita, Desplancq Dominique, Weiss Etienne, Benarafa Charaf, Housset Dominique, Marie Jean-Claude, Frachet Philippe, El-Benna Jamel, Witko-Sarsat Véronique (2019), Cytosolic PCNA interacts with p47phox and controls NADPH oxidase NOX2 activation in neutrophils, in Journal of Experimental Medicine
, 216(11), 2669-2687.
Burgener Sabrina Sofia, Leborgne Nathan Georges François, Snipas Scott J., Salvesen Guy S., Bird Phillip Ian, Benarafa Charaf (2019), Cathepsin G Inhibition by Serpinb1 and Serpinb6 Prevents Programmed Necrosis in Neutrophils and Monocytes and Reduces GSDMD-Driven Inflammation, in Cell Reports
, 27(12), 3646-3656.e5.
Yousefi Shida, Stojkov Darko, Germic Nina, Simon Dagmar, Wang Xiaoliang, Benarafa Charaf, Simon Hans‐Uwe (2019), Untangling “NETosis” from NETs, in European Journal of Immunology
, 49(2), 221-227.
Amini Poorya, Stojkov Darko, Felser Andrea, Jackson Christopher B., Courage Carolina, Schaller André, Gelman Laurent, Soriano Maria Eugenia, Nuoffer Jean-Marc, Scorrano Luca, Benarafa Charaf, Yousefi Shida, Simon Hans-Uwe (2018), Neutrophil extracellular trap formation requires OPA1-dependent glycolytic ATP production, in Nature Communications
, 9(1), 2958-2958.
Martin Katherine R., Pederzoli-Ribeil Magali, Pacreau Emeline, Burgener Sabrina S., Dahdah Albert, Candalh Céline, Lauret Evelyne, Foretz Marc, Mouthon Luc, Lucas Bruno, Thieblemont Nathalie, Benarafa Charaf, Launay Pierre, Witko-Sarsat Véronique (2017), Transgenic Mice Expressing Human Proteinase 3 Exhibit Sustained Neutrophil-Associated Peritonitis, in The Journal of Immunology
, 199(11), 3914-3924.
Serine proteases carried in cytoplasmic granules or lysosomes of granulocytes, monocytes, mast cells, cytotoxic lymphocytes and NK cells contribute to both protective and destructive inflammatory and immune responses. A plethora of functions have been attributed to these proteases such as killing pathogens, inactivating toxins, inducing death of infected or cancer cells, modulating cytokine activity and remodeling extracellular matrix proteins. Much progress has been made in the last decade in understanding the functions of intracellular serpins which inhibit granule proteases of immune cells. Importantly, we have established that Serpinb1 is an essential survival factor of neutrophils (PMNs) through the inhibition of their own granule protease CatG. CatG triggers a form of programmed cell death that has features of apoptosis through activation of caspases and of regulated necrosis with rapid loss of plasma membrane integrity and release of inflammatory mediators. Although we have made significant advances, the exact molecular mechanisms leading to cell death and inflammation in the absence of intracellular serpins remain to be identified. In this project, we will use a combination of targeted (hypothesis-driven) and untargeted (degradomics and phenotype screening) methods to address the knowledge gaps in the mechanisms of CatG-mediated cell death and in the functions of Serpinb1, Serpinb6 and Serpinb9 in vivo. In particular, we have taken the high risk, high reward approach of mouse genetics and developed unique models to directly address these important questions. The proposed project will address the following two aims: Our first specific aim is to identify the mechanisms of cell death mediated by cathepsin G in myeloid cells. We will focus on the biology of myeloid cells and further explore specific cell death pathways associated with PMN and monocyte PCD in vitro. We will use a comparative proteomic approach to identify the CatG-specific degradome in PMNs associated with cell death. The identified targets will be validated by genetic targeting, including CRISPR and Cas9 designer nuclease technology. The second specific aim is to investigate the global and individual functions of the 14 functional serpins encoded in the expanded mouse SerpinB6-SerpinB9-SerpinB1 gene cluster on mouse chromosome 13, for which we generated a complete, yet viable knock-out. We will investigate specific defects of innate and adaptive immune cell development, homeostasis in steady state and inflammatory conditions. We will particularly investigate the cell autonomous and non-cell autonomous functions of invariant NKT cell subsets, which appear to be negatively regulated by SerpinB1 and, potentially, also by other intracellular serpins of this cluster. Finally, we will perform a standardized, comprehensive phenotypic screening of mice lacking all clade B serpin genes on mouse chromosome 13. This explorative analysis will be followed by a second stage testing where identified defects will be functionally investigated for individual serpin genes and their known target proteases. Overall, this multipronged project based on animal genetics will identify and functionally validate novel proteolytic targets associated with programmed cell death and will reveal important pathophysiological mechanisms mediated by granule proteases of immune cells and their inhibitors, which will likely have implications for developmental, inflammatory, infectious and proliferative diseases.