|INBIONET|Infect-ERA| Joint Conference September 14-15, 2016 Queen’s University Belfast
PROGRAMME & ABSTRACT BOOK www.qub.ac.uk/INBIONETInfect-ERAConference
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Contents Welcome Note
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General Information
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Programme
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Abstracts (Infect-ERA )
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Welcome Note Dear participants, On behalf of INBIONET and Infect-ERA we are pleased to welcome you to Queen’s University Belfast for this joint INBIONET/Infect-ERA Conference. This event will be the final conference of the EU funded INBIONET Programme, final seminar for the first joint call of Infect-ERA and the midterm seminar for the second call of the Infect-ERA Programme. This conference is the first time that these two initiatives will meet and, as such, it provides an ideal platform for the 28 represented projects (12 INBIONET projects, 8 projects Infect-ERA JTC1 and 8 projects Infect-ERA JTC2) to present and discuss their results and to initiate new collaborations and new ideas.
We hope you will enjoy this event, meeting your colleagues, having fruitful discussions and making new connections.
Professor José Bengoechea Queen’s University Belfast (INBIONET) ~and~ Martine Batoux The National Agency of Research Projects for Science (Infect-ERA)
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General Information Wi-Fi Free Wi-Fi is available for the duration of the conference by connecting to _The Cloud network at QUB. Visitors from other academic institutions should use “eduroam�. You can get help from The Cloud by visiting their Wi-Fi support pages.
Meals Coffee and tea will be available at refreshment breaks throughout the conference. Lunch will be served in the Canada Room at 1.00pm on both days and Conference Dinner will be served in The Great Hall at 8.00pm on 14th September (welcome reception from 7.30pm). Poster Boards Posters will be available for viewing in the Canada Room/Council Chamber throughout the conference. Poster Presentation will take place from 6.00pm on Wednesday 14th September. Contact In case of important travel issues, e.g. an unavoidable delay in arrival that may impact the start of your presentation or an unpredictable cancellation of attendance on short notice, you may contact the registration desk on +44 [0]28 9097 1643.
Smoking policy Smoking is strictly prohibited within the buildings and grounds of the university except in specifically designated smoking areas. PLEASE NOTE Mobile phones must be switched off during lectures.
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Wednesday 14 September
8.00 - 8.30
Registration
Registration desk Emeleus Lecture Theatre
8.30 - 8.40
Welcome address
Professor Patrick Johnston Vice-Chancellor Queen’s University Belfast
8.40 - 9.00
Opening address
Professor José Bengoechea (Queen’s University Belfast) and Martine Batoux (The National Agency of Research Projects for Science)
9.00-10.30
Session 1
Chair: Professor José Bengoechea Queen’s University Belfast
9.00 - 9.30
Systematic identification of antifungal drug targets by a metabolic network approach (AspMetNet)
Professor Sven Krappmann University Hospital Erlangen, Friedrich-Alexander-University Erlangen-Nürnberg
9.30 - 10.00
Co-infection as a cause of ovarian cancer (CINOCA)
Professor Thomas F. Meyer Max Planck Institute for Infection Biology
10.00 - 10.30
Combining high-throughput and single-cell analyses to study RNA regulators important for the early steps of Campylobacter infection (CampyRNA)
Dr. Miguel Mano University of Coimbra
10.30 - 11.00
Refreshments
Canada Room
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11.00-13.00
Session 2
Chair: Professor José Bengoechea Queen’s University Belfast
11.00 - 11.30
Subversive pro- and antiinflammation trade-offs promote infection by Listeria monocytogenes (PROANTILIS)
Professor Trinad Chakraborty Justus-Liebig University
11.30 - 12.00
Anti-Bacterial Immune Regulation (Abir)
Dr. Jara Joedicke The German Rheumatism Research Centre Berlin, an institute of the Leibniz Association
12.00 - 12.30
Dissection of the interplay between Klebsiella and alveolar macrophages
Sylviane Yoba Queen’s University Belfast
12.30 - 13.00
Early Determinants of DNAVirus Lytic or Latent Infection (eDEVILLI)
Professor Luka Cicin-Sain Helmholtz Centre For Infection Research
13.00 - 14.00
Lunch
Canada Room
14.00 - 15.30
Session 3
Chair: Professor Pavel Kovarik University of Vienna
14.00 - 14.30
IL-1 cytokines in S. pyogenes infection: new view of known players
Marton Janos University of Vienna
14.30 - 15.00
Bacterial interaction with the microvasculature, a target for therapeutic intervention during septicaemia (BactInfectERA)
Dr. Sandrine Bourdoulous The National Health and Medical Research Institute, France
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15.00 - 15.30
Evaluating antiviral properties of viral RNA/DNA Associated proteins across Species Evolution (ERASE)
Dr. Matthias Habja Max Planck Institute of Biochemistry
15.30—6.00
Refreshments
Canada Room
16.00 - 19.00
Session 4
Chair: Professor Pavel Kovarik University of Vienna
16.00 - 16.30
Blockage of IFNgamma signaling by Helicobacter pylori in gastric epithelial cells
Dr. Pau Morey Max Planck Institute for Infection Biology
16.30 - 17.00
TLR2-dependent type I interferon induction by bacteria in human monocytes
Jana Musilova Trinity College Dublin
17.00 - 17.30
Commensalism versus disease? Asymptomatic carriage or urosepsis (The nice bug)
Professor Catharina Svanborg Lund University
17.30 - 18.00
KEYNOTE: Principles and process of evaluation of an article
Professor Erzsébet Nagy Editor of the “anaerobe” journal shares experiences in editorial policy
18.00 - 19.00
Poster Presentation (Infect-ERA)
Canada Room/Council Chamber
19.30 - 20.00
Welcome reception
Adjacent to The Great Hall Queen’s University Belfast
20.00
Conference Dinner
The Great Hall Queen’s University Belfast
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Wednesday 14 SeptemThursday 15 September ber 8.30 - 10.30
Session 1
Chair: Professor Andrew Bowie Trinity College Dublin
8.30 - 9.00
Identification of host factors involved in Hepatitis C Virus assembly and characterization of their potential role in vivo (HCV-ASSEMBLY)
Dr. Jean Dubuisson The National Health and Medical Research Institute, France - Centre for Infection and Immunity of Lille
9.00 - 9.30
From fungal commensalism to pathogenicity: dissection of the colonization-to-infection shift of Candida albicans (FunComPath)
Professor Bernhard Hube Hans Knöll Institute
9.30 - 10.00
Pathophysiology of Klebsiella pneumoniae and Klebsiella rhinoscleromatis infections
Dr. Régis Tournebize Institut Pasteur
10.00 - 10.30
Haploid embryonic stem cell screen to identify host factors essential for highly pathogenic RNA viruses (ESCential)
Professor Ali Mirazimi National Veterinary Institute, Sweden
10.30 - 11.00
Refreshments
Canada Room
11.00 - 13.00
Session 2
Chair: Professor Andrew Bowie Trinity College Dublin
11.00 - 11.30
KEYNOTE: New regulators of innate immune signalling
Professor Paul Moynagh Queen’s University Belfast and The National University of Ireland Maynooth
11.30 - 12.00
Eukaryotic genes in vacuolar pathogens and symbionts - Implications for virulence, metabolism and ecology (EUGENPATH)
Professor Hubert Hilbi Ludwig-Maximilians University Munich
12.00 - 12.30
Investigation of innate immune cell recognition of intestinal helminths
Beatrice Volpe École Polytechnique Fédérale de Lausanne
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12.30 - 13.00
Entamoeba histolyticabacterium interaction and the role of this interaction in intestinal pathogenesis (AMOEBAC)
Professor Serge Ankri Technion - Israel Institute of Technology
13.00 - 14.00
Lunch
Canada Room
14.00 - 15.30
Session 3
Chair: Dr. Régis Tournebize Institut Pasteur
14.00 - 14.30
Analysis of Klebsielladependent control of ubiquitin and ubiquitin-like dependent processes
Kora Przybyszewska Queen’s University Belfast
14.30 - 15.00
Dissecting the bacterial recognition machinery in human cells using haploid genetics and CRISPR-mediated genome engineering (Haplo-Infect)
Dr Ivo Gomperts Boneca Institut Pasteur
15.00 - 15.30
Deciphering the role of type I interferon signaling in Klebsiella pneumoniae infection
Filipe Vasconcelos Queen’s University Belfast
15.30 - 16.00
Refreshments
Canada Room
16.00 - 17.45
Session 4
Chair: Dr. Régis Tournebize Institut Pasteur
16.00 - 16.30
Developing a PP7-based system to capture proteins involved in viral RNA recognition by RLRs
Francisco Dominguez University of St Andrews
16.30 - 17.00
Host factors in hepatitis B virus cccDNA formation as novel antiviral targets and biomarkers identification, preclinical evaluation and impact for liver disease (hepBccc)
Professor Michael Nassal University Hospital Freiburg
17.00 - 17.30
Host and pathogen modulation of innate immune signalling
Professor Andrew Bowie Trinity College Dublin
17.30 - 17.45
Closing Remarks
Professor José Bengoechea Queen’s University Belfast 11
Poster Abstracts—Infect-ERA projects POSTER 1 Joint Transnational Call 1
AspMetNet - Identification and characterization of antifungal drug targets in Aspergillus fumigatus AFFILIATIONS Anna-Maria Dietl1, Jorge Amich2, Sixto Leal3, Ulrike Binder4, Nir Osherov6, Hubertus Haas1 1
Div. of Molecular Biology/Biocenter, Medical University of Innsbruck; 2IZKF Forschergruppe für Experimentelle Stammzelltransplantation, Würzburg; 3Dept. of Ophthalmology, Case Western Reserve University, Cleveland, Ohio; 4 Div. of Hygiene & Medical Microbiology, Medical University of Innsbruck; 6Dept. of Human Microbiology, Sackler School of Medicine, Tel Aviv ABSTRACT Aspergillus fumigatus is the most prevalent airborne fungal pathogen causing invasive fungal infections in immunosuppressed individuals. Limitations in antifungal therapy arise from non-specific symptoms of infection, poor diagnostics and comparatively few options for treatment. The aim of this study is to explore the metabolism of A. fumigatus on a comprehensive scale as essential virulence determinant to generate a collection of A. fumigatus strains with a focus on primary metabolism to target fungal pathways that are absent in mammals. Based on the annotated genome of A. fumigatus, metabolic network reconstruction served to identify fungal-specific pathways and key reactions. Predictions for unique enzymes resulted in a candidate list of genes, the inactivation of which is likely to result in an auxotrophic phenotype. We identified four essential biosynthetic pathways in A. fumigatus: (i) biosynthesis of the amino acid histidine, (ii) biosynthesis of the cofactor siroheme, which is essential for sulphate and nitrate assimilation, (iii) biosynthesis of the vitamin riboflavin and (iiii) biosynthesis of the vitamin pantothenic acid. The virulence potential of the generated auxotrophic mutant strains was then analyzed in various host niches. PROJECT PARTNER Prof. Hubertus Haas - Medical University of Innsbruck (AT) COORDINATOR Prof. Sven Krappmann - University Hospital Erlangen, Friedrich-Alexander-University Erlangen-Nürnberg (DE)
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POSTER 2
Joint Transnational Call 1
Eugenpath - Bipartite metabolism in the intracellular pathogenic bacterium Legionella pneumophila AFFILIATIONS Ina Häuslein (Technical University of Munich), Christian Manske (Max von PettenkoferInstitute), Werner Goebel (Max von Pettenkofer-Institute), Wolfgang Eisenreich (Technical University of Munich) and Hubert Hilbi (University of Zßrich) ABSTRACT Intracellular pathogens are responsible for several life-threatening respiratory diseases including Legionnaires' disease caused by Legionella pneumophila The pathogen replicates in a specific vacuole (LCV) of host cells e.g. macrophages where it resists degradation by the host involving control by multiple factors. There is now increasing evidence that the adaptation process also crucially depends on modulating the basic metabolic pathways and fluxes of the pathogens. To analyze these complex metabolic processes in Legionella pneumophila, we have exploited stable isotope labeling experiments for a detailed analysis of substrate usage and downstream metabolic pathways leading to protein-derived amino acids, cell wall constituents and some low molecular weight metabolites. 13C-Enrichments and positional isotope distributions in multiple metabolic products were analyzed by GC/MS and/or NMR spectroscopy to provide global 13C-enrichments and positional 13C-distributions. On this basis, relative rates of substrate usage, metabolic pathways and fluxes were obtained. The data reflect that L. pneumophila simultaneously metabolizes serine, glucose and glycerol, however at different rates depending on the growth phase. The metabolic network is characterized by a bipartite topology, where one set of substrates is used for energy generation and a second set is used for anabolic purposes, i.e. cell wall or protein biosynthesis. Specifically, L. pneumophila uses serine predominantly for energy generation in the TCA cycle while glucose and glycerol are used in gluconeogenetic reactions and the pentose phosphate pathway under in vitro and in vivo studies. This principle of a bipartite metabolism could benefit survival of the intracellular pathogen. Similar results were obtained with Listeria monocytogenes, Chlamydia trachomatis and Coxiella burnetii. Thus, bipartite metabolism could therefore represent a general concept of intercellular life style. PROJECT PARTNER Prof. Hubert Hilbi - Ludwig-Maximilians University Munich (DE) COORDINATOR Prof. Hubert Hilbi - Ludwig-Maximilians University Munich (DE)
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POSTER 3
Joint Transnational Call 1
hepBCCC - A targeted RNAi screen using a high-throughput infectious model system uncovers Glypican GPC5 as a host factor for hepatitis B and D virus entry AFFILIATIONS Eloi R. Verrier1,2, Che C. Colpitts1,2, Charlotte Bach1,2, Laura Heydmann1,2, Amélie Weiss3, Mickaël Renaud3, Sarah Durand1,2, François Habersetzer4, Georges Abou-Jaoudé5, David Durantel6,7, Maria M. Lopez Ledesma8, David J. Felmlee1,2, Magali Soumillon9, Tom Croonenborghs9, Nathalie Pochet9, Michael Nassal10, Catherine Schuster1,2, Laurent Brino3, Camille Sureau5, Mirjam B. Zeisel1,2, Thomas F. Baumert1,2,4 1
Inserm, U1110, Institut de Recherche sur les Maladies Virales et Hépatiques, 67000, Strasbourg, France; 2Université de Strasbourg, 67000 Strasbourg, France; 3IGBMC, Plateforme Criblage Haut-débit, 67404 Illkirch, France; 4Pôle Hépato-digestif, Institut Hospitalo-Universitaire, 67000 Strasbourg, France. 5INTS, Laboratoire de Virologie Moléculaire, 75015 Paris, France. 6Inserm U1052, Centre de Recherche en Cancérologie de Lyon, Lyon, France. 7Université de Lyon, Lyon, France. 8Catedra de Virologıa, Facultad de Farmacia y Bioquımica, Universidad de Buenos Aires, Buenos Aires, Argentina. 9Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, MA, USA. 10 Department of Internal Medicine II/Molecular Biology, University Hospital Freiburg, Freiburg, Germany. ABSTRACT Chronic hepatitis B and D infections are major causes of liver disease and hepatocellular carcinoma worldwide. Efficient therapeutic approaches for cure are absent. Sharing the same envelope proteins, hepatitis B virus (HBV) and hepatitis D virus (HDV) use the sodium taurocholate co-transporting polypeptide (NTCP, a bile acid transporter) as a receptor to enter the hepatocytes. However, the detailed mechanisms of the viral entry process are still poorly understood. Here, we established a high-throughput infectious cell culture model enabling functional genomics of HDV entry and infection. Using a targeted RNAi entry screen we identified glypican 5 (GPC5) as a common host cell entry factor for HBV and HDV. These findings advance our understanding of virus cell entry and open new avenues for curative therapies. Since glypicans have been shown to play a role in the control of cell division and growth regulation, virus-GPC5 interactions may also play a role for pathogenesis of virus-induced liver disease and cancer. PROJECT PARTNER Prof. Thomas Baumert - University of Strasbourg (FR) COORDINATOR Prof. Nassal Michael - University Hospital Freiburg (DE) 14
POSTER 4
Joint Transnational Call 1
PROANTILIS - Identification of RNAs and ribosomal proteins associated to SecA2 in Listeria monocytogenes AFFILIATIONS Teubner, Lisa; Abu Mraheil, Mobarak; Chakraborty, Trinad; all from Institute for Medical Microbiology JLU Giessen, Germany ABSTRACT Listeria monocytogenes (Lm) is a food-borne pathogen that is the cause of listeriosis in humans. Recently, it was shown that the recognition of listerial secreted RNAs is mediated by the cytosolic receptors RIG-I and MDA5 which lead to the induction of IFN-β in macrophages. The secretion pathway for RNA in Lm is not yet well-understood. The general secretion (“Sec”) pathway is considered to be the main system for protein secretion in Lm. Lm also encodes for an additional SecA protein, the SecA2. Mutants lacking SecA2 (ΔsecA2) have reduced amounts of secreted nucleic acids and lowered IFN-β induction levels as compared to the wildtype, suggesting a role of SecA2 in the active secretion of nucleic acids in Lm. Strep-tagged SecA2 protein was isolated from complemented ΔsecA2 cells and associated RNAs from purified SecA2 protein were sequenced by RNA-Seq. The sequence data reveal the existence of several small non-coding RNAs (sRNA), which are able individually to induce a higher IFN-β expression in transfected macrophages. Moreover, mass spectrometry analyses have shown an enrichment of ribosomal proteins in a purified SecA2 protein solution. Co-sedimentation experiments confirmed the binding of SecA2 to listerial ribosomes. Further investigations are needed to support our hypothesis that SecA2 may assist a cotranslational initiation of secretion processes in Lm as part of a putative complex composed of ribosomes and RNAs. PROJECT PARTNER Prof. Trinad Chakraborty - Justus-Liebig University (DE) COORDINATOR Prof. Trinad Chakraborty - Justus-Liebig University (DE)
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POSTER 5
Joint Transnational Call 2
BactInfectERA - Characterization of the bacterial and host genes involved in meningococcal disease initiation and progression AFFILIATIONS Nussbaumer, Thomasa; Join-Lambert Olivierb,c,d,e; Le Guennec Loïcf,g,h; Chosidow Anaïsf,g,h ; Gouge Annef,g,h; Segard-Bouzinba Haniaaf,g,h Izac Brigittef,g,h; Letourneur Franckf,g,h; Pelissier Philippei; Morand Philippef,g,h,j; Coureuil Mathieub,c,d; Nassif Xavier b,c,d,e; Hidalgo Andrésk,l; Rattei Thomasa and Bourdoulous Sandrinef,g,h CUBE - Division of Computational Systems Biology, Vienna, Austria a; Inserm, unité U1151, Institut-Necker-Enfants-Malades, Paris, Paris, Franceb; CNRS, UMR 8253, Paris, Francec ; Université Paris Descartes, Sorbonne Paris Cité, Faculté de Médecine, Paris, Franced ; Assistance Publique – Hôpitaux de Paris, Hôpital Necker Enfants Malades, Paris, Francee ; Inserm, U1016, Institut Cochin, Paris, Francef ; CNRS, UMR8104, Paris, Franceg ; Université Paris Descartes, Sorbonne Paris Cité, Franceh ; Service de Chirurgie Reconstructrice et Plastique, Fondation Hôpital Saint Joseph, Parisi; Assistance Publique – Hôpitaux de Paris, Hôpital Cochin, Paris, Francej; Fundación Centro Nacional de Investigaciones Cardiovasculares , Madrid , Spaink; Institute for Cardiovascular Prevention, Ludwig-Maximilians University , Munich , Germanyl ABSTRACT Neisseria meningitidis (meningococcus) is an invasive bacterial pathogen responsible for rapidly progressing fatal septic shock and meningitis. It is an obligate human Gram-negative bacterium that normally resides in the nasopharyngeal mucosa of ~10% of the human population without affecting the host. Pathology is initiated when meningococci translocate from the throat into the bloodstream. Once in the blood flow, specific interaction of this bacterium with human endothelia enables bacterial access to secondary anatomical sites, which promotes much of the morbidity and mortality associated with neisserial infections. Major contribution in the understanding of this intricate host-pathogen interaction were made in vitro by identifying key bacterial factors and host cell receptors required for the initial adhesion of N. meningitidis to human endothelial cells and the triggering of host signalling events leading to endothelial cells disruption. However, the mechanisms and consequences in vivo of such colonization on vascular remodelling and alteration now remain to be explored. A recent in vivo model using severe combined immunodeficient mice grafted with human skin, has been developed, in which circulating meningococci specifically colonize the human endothelia contained within the graft and induce vascular lesions similar to those described in patients. Here, we take advantage of this novel and pertinent in vivo model of meningococcal infection and of novel next-generation deep-sequencing platforms to conduct an unbiased and global characterization of the associated gene expression changes in both the pathogen and the host. Among the differentially expressed genes, we find important pathogenic virulence factors (e.g. Maf family of polymorphic toxins, Type IV 16
pili). Whereas in the host, colonization was associated with transcriptional reorientation of the endothelial cells toward inflammatory responses (NF-κB pathway, cytokines), adhesion molecule expression (ICAM1, VCAM1, E selectin), thrombosis and fibrinolysis (PLAT, PLAU, PLAUR, ...) or matrix degradation (MMP, ADAMTS). The induction of selective pathways (NF-κB) and genes (IL-8) were analyzed confirming these findings. Of most interest, several other genes involved in maintaining vascular homeostasis, endothelial junction integrity were regulated and will be investigated further. Taking advantage of the complementary nature of the data (host & pathogen) and connecting them to the knowledge from public sequence and pathway databases, now we use bioinformatics and mathematical methods to identify strain-specific genes involved in pathogenicity, establish Host-Pathogen proteinprotein interaction networks and perform a modelling of bacterial metabolic networks.
PROJECT PARTNER Prof. Thomas Rattei - University of Vienna (AT) COORDINATOR Dr. Sandrine Bourdoulous - INSERM (FR)
POSTER 6 Joint Transnational Call 2
BactInfectERA - Tn-seq identification of meningococcal factors required for in vitro and in vivo colonization of human cells AFFILIATIONS Capel Ea,b; Zomer ALc; Barnier JPa,b,d; Nussbaumer Te; Bole Cf; Izac Bb,g; Frapy Ea,b; Meyer Ja,b; Euphrasie Da,b; Bouzinba-Ségard Hb,g; Bille Ea,b,d; Jamet Aa,b,d,h; Cavau Aa,b; Letourneur Fb,g; Bourdoulous Sb,g; Rattei Te; Join-Lambert Oa,b,d; Nassif Xa,b,d and Coureuil Ma,b. a
Institut Necker Enfants-Malades, INSERM U1151, Equipe 11, Paris, France; bUniversité Paris Descartes; Sorbonne Paris Cité, Faculté de Médecine, Paris, France; cDepartment of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, The Netherlands; dAssistance Publique – Hôpitaux de Paris, Hôpital Necker Enfants Malades, Paris, France; eCUBE - Division of Computational Systems Biology, Dept. of Microbiology and Ecosystem Science, University of Vienna, Austria; fPlateforme génomique de l'Institut Imagine, Hôpital Necker, Paris, France; gINSERM U1016, Institut Cochin, CNRS UMR8104, Paris, France; hUnidade de Microbiologia Molecular e Infecção, Instituto de Medicina Molecular, Lisbon, Portugal. ABSTRACT Neisseria meningitidis is a leading cause of bacterial meningitis and septicemia affecting infants and adults worldwide. N. meningitidis is also a common inhabitant of the human nasopharynx and, as such, is highly adapted to its niche. During bacteremia, N. meningitidis 17
gains access to the blood compartment where it adheres to endothelial cells of blood vessels causing dramatic vascular damages. Colonization of the nasopharyngeal niche and communication with the different human cell types is a major aspect of the N. meningitidis life cycle that is poorly understood. Here, highly saturated random transposon insertion libraries of N. meningitidis were engineered and the fitness of mutations during (i) routine growth, (ii) in vitro colonization of endothelial or epithelial cells in a flow device, and (iii) in vivo colonization of human vessels in a human skin grafted mice model was assessed, using Tn-seq analysis. This allowed the identification of genes essential for bacterial growth and genes specifically required for host cell colonization. In addition, after having identified the small non-coding RNAs (sRNA) located in intergenic regions, the phenotypes associated with mutation in those sRNAs were defined. 383 genes and 8 intergenic regions containing sRNA candidates were identified as essential for growth while 288 genes and 33 intergenic regions containing sRNA candidates were specifically required for host cell colonization. Besides the role of type IV pili for colonization, we observed a metabolic reorientation towards biosynthesis of cellular components, which is consistent with greater proliferation, and identified genes important for colonization of blood vessels in vivo. PROJECT PARTNER Dr. Sandrine Bourdoulous - INSERM (FR) COORDINATOR Dr. Sandrine Bourdoulous - INSERM (FR)
POSTER 7 Joint Transnational Call 2
CampyRNA – Combining high-throughput and single-cell analyses to study RNA regulators important in the early steps of Campylobacter infection AFFILIATIONS Mona Alzheimer1, Carmen Aguilar2, Ana Rita Cruz3, Sarah L. Svensson1, Kristine Schauer4, Miguel Mano3, Ana Eulalio2, Cynthia M. Sharma1,2 1
Research Center for Infectious Diseases (ZINF), University of Würzburg, Würzburg, Germany,2 Institute of Molecular Infection Biology (IMIB), University of Würzburg, Würzburg, Germany,3 Center for Neuroscience and Cell Biology (CNC), University of Coimbra, Coimbra, Portugal,4 Centre National de la Recherche Scientifique (CNRS) / Institut Curie, Paris, France
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ABSTRACT Bacterial infections are characterized by an active interplay between the virulence determinants of the pathogen and the host response. This process is accompanied by major gene expression changes of both pathogen and host. Non-coding RNAs (ncRNAs) have been increasingly recognized as global regulators of gene expression in both eukaryotes and prokaryotes. For example, host microRNAs (miRNAs) can act as central post-transcriptional regulators of mRNAs and have been shown to play crucial roles in controlling immune response pathways during infection. In bacteria, small RNAs (sRNAs) act as antisense RNAs or directly modulate protein activity and control stress response and virulence factor expression in many human pathogens. In our project, we aim to globally investigate the role of such ncRNAs during the early steps of Campylobacter jejuni infections. The zoonotic pathogen C. jejuni is currently the most common cause of bacterial food-borne gastroenteritis and can also lead to several secondary autoimmune disorders. So far, its mechanisms of pathogenesis and colonization of different hosts are still poorly understood. We are focused on the interplay between C. jejuni and human cells, with a special interest in the identification and characterization of host and pathogen ncRNAs and their targets, as well as in the mechanisms whereby they control and restrict infection, particularly during adhesion and invasion of epithelial cells. For this purpose, we are combining several genome-wide approaches, such as RNA-seq, Tnseq, and high-content screening, with single-cell microscopy techniques based on micropatterned cells. Based on these global approaches we will select bacterial sRNAs and host miRNAs for further functional characterization. We have established experimental conditions to study the early steps of C. jejuni infection. Moreover, we have constructed a high-density transposon mutant library for C. jejuni. Infection of human epithelial cells with this C. jejuni mutant library and analysis of mutant pools before and after infection by deep sequencing have revealed several mutants defective for adherence and invasion with transposon insertions in previously unidentified factors that may aid bacterial fitness during the course of infection. Moreover, we have performed a pilot screening with a subset of human miRNA mimics, which revealed miRNAs controlling the early steps of interaction of C. jejuni with host cells. Ongoing work encompasses the screening of a genome-wide library of miRNA mimics (2,588 mature miRNAs). Further studies will validate the role of these ncRNAs as well as their potential targets. The identification of potential novel host and pathogen factors that influence the course of infection may provide the opportunity for alternative approaches for antimicrobial therapy. Finally, these global approaches will also allow for investigation of host-microbial interactions of additional human pathogens. PROJECT PARTNER Dr. Cynthia Mira Sharma - Research Center for Infectious Diseases, University of WĂźrzburg (DE) COORDINATOR Dr. Cynthia Mira Sharma - Research Center for Infectious Diseases, University of WĂźrzburg (DE) 19
POSTER 8
Joint Transnational Call 2
FunComPath – Investigating the interactions between Candida albicans and lactobacilli using a commensal in vitro gut model AFFILIATIONS Antonia Dräger1,2, Katja Graf1, Bernhard Hube1,2,3 1
Department of Microbial Pathogenicity Mechanisms, HKI, Jena, Germany, Center for Sepsis Control and Care, University Hospital Jena, Germany, 3 Friedrich Schiller University, Jena, Germany 2
ABSTRACT Candida albicans is both, a harmless commensal on mucosal surfaces and the most common cause of life-threatening nosocomial fungal infections. C. albicans infections are generally caused by endogenous strains which colonize mucosal surfaces. In both stages, as a commensal and as a pathogen, C. albicans not only interacts with the host but also with other members of the microbial flora. Thereby the removal or imbalance of the bacterial microbiota is a significant predisposing factor for disseminated Candida infections. Our aim is to investigate the commensal-to-pathogen shift of C. albicans and the role of non-pathogenic bacteria during or in preventing this shift. Therefore we established a two cell line based commensal in vitro gut model. Using this model we were able to show a time - and dose dependent protective effect of different lactobacilli strains against C. albicans induced cell damage. This protective effect required viable lactobacilli and was not related to a competition for adhesion sites. Moreover, protection required the presence of host cells. Next, similar to the clinical situation, an antibiotic treatment will be used to remove the protective lactobacilli and thereby induce a shift from the commensal to a pathogenic state of C. albicans.Transcriptional profiling will help us to identify genes which are involved in the commensal-to-pathogen shift of C. albicans and will help to elucidate the mechanisms behind the protective effect of lactobacilli. All in all, the results of this project will contribute to a better understanding of the interactions of different microorganisms within the human host and the impact of these interactions on disease development. This could lead to novel prophylactic and therapeutic approaches. PROJECT PARTNER Prof. Bernhard Hube - Hans Knöll Institute (DE) COORDINATOR Prof. Bernhard Hube - Hans Knöll Institute (DE)
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POSTER 9 Joint Transnational Call 2
The nice bug – Specific IRF-7 agonism in UPEC strains drives human disease - Fimbriae reprogram host gene expression AFFILIATIONS Ines Ambite1, Jenny Gronberg-Hernandez1, Christoph Stork2, Bela Koves1, Jaroslaw Zdziarski3, Ulrich Dobrindt2,3, Bjorn Wullt1, Catharina Svanborg1 1
Dept of Microbiology, Immunology and Glycobiology, Inst of Laboratory Medicine, Lund University, Sweden. 2 Inst for Hygiene, University of Munster, Germany. 3 Inst for Molecular Biology of Infectious Diseases, University of Wurzburg, Germany. ABSTRACT Pathogens need multiple virulence factors for tissue attack and may evolve into commensals, if virulence genes are attenuated. Here we describe the surprising observation that virulence can be regained, solely by the acquisition of a functional pap gene cluster. The identified mechanism is reprogramming of host gene expression, in susceptible human hosts. By acting as specific agonists for the transcription factor IRF-7, P fimbriae triggered an inflammatory host response that resembled acute pyelonephritis. The effects of type 1 fimbriae were radically different, with attenuation of gene expression. It is conceivable that uropathogenic E. coli have developed specific IRF-7 agonism to drive hyper inflammatory responses and promote efficient dissemination. This strong effect of a functional pap gene cluster suggests that certain virulence factors are potent enough to trigger a disease response, without a pathogenic strain background. Such virulence factors are interesting targets in attempts to reduce pathology caused by infection. PROJECT PARTNER Prof. Catharina Svanborg - Lund University (SE)
COORDINATOR Prof. Catharina Svanborg - Lund University (SE)
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Notes
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THANKS TO OUR SPONSORS
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