Bacterial virulence; Infection; 3R principles; Non-mammalian and mammalian models; Library design; Relational database; Pathogenesis; Microfluidic-microscopy systems; Macrophages; Anti-infectiva
Garcia-Lopez Amparo, Tessaro Francesca, Jonker Hendrik R. A., Wacker Anna, Richter Christian, Comte Arnaud, Berntenis Nikolaos, Schmucki Roland, Hatje Klas, Petermann Olivier, Chiriano Gianpaolo, Perozzo Remo, Sciarra Daniel, Konieczny Piotr, Faustino Ignacio, Fournet Guy, Orozco Modesto, Artero Ruben, Metzger Friedrich, Ebeling Martin, Goekjian Peter, Joseph Benoît, Schwalbe Harald, Scapozza Leonardo (2018), Targeting RNA structure in SMN2 reverses spinal muscular atrophy molecular phenotypes, in Nature Communications
, 9(1), 2032-2032.
Trofimov Valentin, Kicka Sébastien, Mucaria Sabrina, Hanna Nabil, Ramon-Olayo Fernando, Del Peral Laura Vela-Gonzalez, Lelièvre Joël, Ballell Lluís, Scapozza Leonardo, Besra Gurdyal S., Cox Jonathan A. G., Soldati Thierry (2018), Antimycobacterial drug discovery using Mycobacteria-infected amoebae identifies anti-infectives and new molecular targets, in Scientific Reports
, 8(1), 3939-3939.
Ouertatani-Sakouhi Hajer, Kicka Sébastien, Chiriano Gianpaolo, Harrison Christopher F., Hilbi Hubert, Scapozza Leonardo, Soldati Thierry, Cosson Pierre (2017), Inhibitors of Mycobacterium marinum virulence identified in a Dictyostelium discoideum host model, in PLOS ONE
, 12(7), e0181121-e0181121.
Delincé Matthieu J., Bureau Jean-Baptiste, López-Jiménez Ana Teresa, Cosson Pierre, Soldati Thierry, McKinney John D. (2016), A microfluidic cell-trapping device for single-cell tracking of host–microbe interactions, in Lab on a Chip
, 16(17), 3276-3285.
Delincé Matthieu J, Bureau Jean-Baptiste, López-Jiménez Ana Teresa, Cosson Pierre, Soldati Thierry, McKinney John D (2016), A microfluidic cell-trapping device for single-cell tracking of host-microbe interactions., in Lab on a chip
, 16(17), 3276-85.
Slepikas Liudas, Chiriano Gianpaolo, Perozzo Remo, Tardy Sébastien, Kranjc Agata, Patthey-Vuadens Ophélie, Ouertatani-Sakouhi Hajer, Kicka Sébastien, Harrison Christopher F, Scrignari Tiziana, Perron Karl, Hilbi Hubert, Soldati Thierry, Cosson Pierre, Tarasevicius Eduardas, Scapozza Leonardo (2016), In Silico Driven Design and Synthesis of Rhodanine Derivatives as Novel Antibacterials Targeting the Enoyl Reductase InhA., in Journal of medicinal chemistry
Harrison Christopher F, Kicka Sébastien, Kranjc Agata, Finsel Ivo, Chiriano Gianpaolo, Ouertatani-Sakouhi Hajer, Soldati Thierry, Scapozza Leonardo, Hilbi Hubert (2015), Adrenergic antagonists restrict replication of Legionella., in Microbiology (Reading, England)
, 161(7), 1392-406.
Harrison Christopher F, Chiriano Gianpaolo, Finsel Ivo, Manske Christian, Hoffmann Christine, Steiner Bernhard, Kranjc Agata, Patthey-Vuadens Ophelie, Kicka Sébastien, Trofimov Valentin, Ouertatani-Sakouhi Hajer, Soldati Thierry, Scapozza Leonardo, Hilbi Hubert (2015), Amoebae-Based Screening Reveals a Novel Family of Compounds Restricting Intracellular Legionella pneumophila., in ACS infectious diseases
, 1(7), 327-38.
Finsel Ivo, Hilbi Hubert (2015), Formation of a pathogen vacuole according to Legionella pneumophila: how to kill one bird with many stones., in Cellular microbiology
, 17(7), 935-50.
Kicka Sébastien, Trofimov Valentin, Harrison Christopher, Ouertatani-Sakouhi Hajer, McKinney John, Scapozza Leonardo, Hilbi Hubert, Cosson Pierre, Soldati Thierry (2014), Establishment and validation of whole-cell based fluorescence assays to identify anti-mycobacterial compounds using the Acanthamoeba castellanii-Mycobacterium marinum host-pathogen system., in PloS one
, 9(1), 87834-87834.
Hoffmann Christine, Harrison Christopher F, Hilbi Hubert (2014), The natural alternative: protozoa as cellular models for Legionella infection., in Cellular microbiology
, 16(1), 15-26.
Rothmeier Eva, Pfaffinger Gudrun, Hoffmann Christine, Harrison Christopher F., Grabmayr Heinrich, Repnik Urška, Hannemann Mandy, Wölke Stefan, Bausch Andreas R., Griffiths Gareth, Müller-Taubenberger Annette, Itzen Aymelt, Hilbi Hubert (2013), Activation of Ran GTPase by a Legionella Effector Promotes Microtubule Polymerization, Pathogen Vacuole Motility and Infection, in PLoS Pathogens
, 9(9), e1003598.
Dias Marco, Blanc Cédric, Thazar-Poulot Nelcy, Ben Larbi Sabrina, Cosson Pierre, Letourneur François (2013), Dictyostelium ACAP-A is an ArfGAP involved in cytokinesis, cell migration and actin cytoskeleton dynamics., in Journal of cell science
, 126(Pt 3), 756-66.
Wakamoto Yuichi, Dhar Neeraj, Chait Remy, Schneider Katrin, Signorino-Gelo François, Leibler Stanislas, McKinney John D (2013), Dynamic persistence of antibiotic-stressed mycobacteria., in Science (New York, N.Y.)
, 339(6115), 91-5.
Harrison Christopher F., Kicka Sébastien, Trofimov Valentin, Berschl Kathrin, Ouertatani-Sakouhi Hajer, Ackermann Nikolaus, Hedberg Christian, Cosson Pierre, Soldati Thierry, Hilbi Hubert (2013), Exploring Anti-Bacterial Compounds against Intracellular Legionella, in PLoS ONE
, 8(9), e74813.
Arafah Sonia, Kicka Sébastien, Trofimov Valentin, Hagedorn Monica, Andreu Nuria, Wiles Siouxsie, Robertson Brian, Soldati Thierry (2013), Setting up and monitoring an infection of Dictyostelium discoideum with mycobacteria., in Methods in molecular biology (Clifton, N.J.)
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Santi Isabella, Dhar Neeraj, Bousbaine Djenet, Wakamoto Yuichi, McKinney John D (2013), Single-cell dynamics of the chromosome replication and cell division cycles in mycobacteria., in Nature communications
, 4, 2470-2470.
Finsel Ivo, Ragaz Curdin, Hoffmann Christine, Harrison Christopher F, Weber Stephen, van Rahden Vanessa A, Johannes Ludger, Hilbi Hubert (2013), The Legionella effector RidL inhibits retrograde trafficking to promote intracellular replication., in Cell host & microbe
, 14(1), 38-50.
Hilbi Hubert, Hoffmann Christine, Harrison Christopher F (2011), Legionella spp. outdoors: colonization, communication and persistence., in Environmental microbiology reports
, 3(3), 286-96.
IDENTIFICATION AND CHARACTERIZATION OF NOVEL ANTIBACTERIAL COMPOUNDS USING PROTOZOAN HOSTSSUMMARYThe general aim of this project is to discover and characterize novel antibacterial compounds with the ultimate goal to be used to prevent or treat bacterial infections. We will focus primarily on compounds inhibiting bacterial virulence, and secondarily on compounds boosting host defence or acting as antibiotics. The endpoint of the project is the identification of antibacterial compounds with the characteristics of validated hits, namely concentration-dependent antibacterial activity, an IC50 < 10 uM, 2 log bacteria-host (protozoan and mammalian) selectivity and a first structure-activity relationship (SAR).Bacterial infections are a major public health problem worldwide. The threat is aggravated by the evolution of drug resistance, a paucity of new antibiotics, and the emergence of previously unrecognized infectious agents. For these reasons, the identification of novel antibacterial drugs is a pressing issue. The screening of small molecule chemical libraries is the strategy of choice to identify novel compounds. A detailed understanding of bacterial virulence and the development of powerful screening tools are prerequisites to identify this new type of drugs.In view of the overall failure of target-based screening campaigns, it is essential to develop novel phenotype-based screens such as the ones proposed here based on pathogen-host interactions. Academia should play a leading role in exploring this research avenue. Our project focuses on the first steps of drug-discovery: the development of adequate screening tests, the isolation of hits and the early characterization of structure-activity relationships. For the primary screens, we will use amoebae as non-mammalian infection models because they are easy to handle, robust, and cost-effective. Dictyostelium discoideum and/or Acanthamoeba castellanii will be used as hosts in combination with the following human relevant pathogens: i) Klebsiella pneumoniae, Pseudomonas aeruginosa (Subproject B), ii) Legionella pneumophila (Subproject C), iii) Mycobacterium spp. (Subproject D). For the validation of the hits resulting from the protozoan screens in a mammalian system, a macrophage-based microfluidic-microscopy system will be developed to study the interaction of immune phagocytes with the above-mentioned pathogens at the single cell level (Subproject E). The experimental screening will make use of a highly diverse low size (5000 compounds) chemical library, which targets pathways essential for bacterial virulence and host resistance, and which covers the chemical and biological space of the compounds available for screening. The library will be designed starting from 8 millions compounds using ligand-based virtual screening (Subproject A), combining the knowledge on ligands known from the literature to interact with relevant host-pathogen pathways and the information from the unique scaffolds covering the chemical space. The compounds annotated with molecular descriptors and the results from the phenotypic screening will be deposited into an on-line database (Shared Results Database) to ensure the flow of the results within the different partners of this Sinergia project and to allow data mining and SAR studies. Three of the participants of this application have long-standing experience with non-mammalian host models and one with mammalian phagocytes to study the virulence of different bacterial pathogens. One participant is a specialist in pharmaceutical chemistry with long-standing experience in virtual screening and library design. This Sinergia team represents the necessary expertise in experimental microbiology and pharmaceutical chemistry, and thus, is in an ideal position to explore this novel approach to develop new anti-bacterial drugs. The long-term vision is that compounds identified in this project will be taken forward into a lead optimization phase that may eventually produce novel clinically useful anti-infective compounds. Moreover, the compounds identified will serve as valuable tools to study cellular microbiology of human pathogens.The specific aims of this proposal are to1. Develop robust and simple assays to: i) screen for new antibacterial compounds using amoebae hosts and representative bacterial pathogens, and ii) perform real-time analysis of bacteria-phagocyte interactions. 2. Design and screen a highly diverse low size chemical library to identify novel antibacterial compounds with hit characteristics in protozoan systems, which are validated in mammalian systems.3. Perform data mining and characterize candidate antibacterial compounds mechanistically. Expected valueWe expect to provide a proof of concept on a series of points. First, we will demonstrate that phenotypic screening in a simple system allows cost-effective and rapid identification of primary antibacterial compounds. Second, we expect to efficiently filter out the usual frequent hitters that can otherwise only be eliminated in laborious cellular and animal tests, such as membrane-active and cytotoxic compounds. Third, we will explore the phylogenetic conservation of bacterial virulence and host resistance mechanisms and at the same time advance the cause of 3R-compliant research aiming at reducing the need for animal experiments. Fourth, we anticipate identifying not only antibacterial compounds but also “boosters” of the cell-intrinsic defence mechanisms. Fifth, we expect to identify compounds that will advance the cellular and molecular understanding of host-pathogen interactions.