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1. Insights in the molecular mechanisms of an azole stress adapted laboratory-generated Aspergillus fumigatus strain

Author

Patricia Flamant, Isabelle Mouyna, Samantha Gozel, Alix T. Coste, Dominique Sanglard, Frédéric Lamoth, Josie E. Parker, Daniel Bachmann, Marion Aruanno, Lausanne University Hospital, Université de Lausanne = University of Lausanne (UNIL), Aspergillus, Institut Pasteur [Paris] (IP), Swansea University, and This work was supported by the Swiss National Science Foundation (SNSF Ambizione-Score, grant number: PZ00P3 161140) and by the Santos-Suarez foundation.

Subjects
Azoles, Antifungal Agents, ATP-binding cassette transporter, Aspergillus fumigatus, chemistry.chemical_compound, MESH: HMGB1 Protein, Cytochrome P-450 Enzyme System, MESH: Reverse Transcriptase Polymerase Chain Reaction, MESH: Azoles, MESH: Ku Autoantigen, HMGB1 Protein, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, chemistry.chemical_classification, 0303 health sciences, Ergosterol, biology, MESH: Real-Time Polymerase Chain Reaction, Reverse Transcriptase Polymerase Chain Reaction, MESH: Voriconazole, General Medicine, isavuconazole milbemycine, Sterols, Infectious Diseases, MESH: CCAAT-Binding Factor, MESH: Cytochrome P-450 Enzyme System, MESH: ATP-Binding Cassette Transporters, MESH: Fungal Proteins, Efflux, MESH: Aspergillus fumigatus, efflux pumps, transporters, Real-Time Polymerase Chain Reaction, Gas Chromatography-Mass Spectrometry, MESH: Drug Resistance, Fungal, Microbiology, resistance, Fungal Proteins, 03 medical and health sciences, Drug Resistance, Fungal, Ku Autoantigen, Transcription factor, 030304 developmental biology, 030306 microbiology, MESH: Transcriptome, Cell Membrane, Transporter, MESH: Antifungal Agents, biology.organism_classification, posaconazole, Major facilitator superfamily, MESH: Gas Chromatography-Mass Spectrometry, CCAAT-Binding Factor, chemistry, MESH: Sterols, Azole, ATP-Binding Cassette Transporters, Voriconazole, erogsterol, Transcriptome, MESH: Cell Membrane
Abstract

Aspergillus fumigatus is the main cause of invasive aspergillosis, for which azole drugs are the first-line therapy. Emergence of pan-azole resistance among A. fumigatus is concerning and has been mainly attributed to mutations in the target gene (cyp51A). However, azole resistance may also result from other mutations (hmg1, hapE) or other adaptive mechanisms. We performed microevolution experiment exposing an A. fumigatus azole-susceptible strain (Ku80) to sub-minimal inhibitory concentration of voriconazole to analyze emergence of azole resistance. We obtained a strain with pan-azole resistance (Ku80R), which was partially reversible after drug relief, and without mutations in cyp51A, hmg1, and hapE. Transcriptomic analyses revealed overexpression of the transcription factor asg1, several ATP-binding cassette (ABC) and major facilitator superfamily transporters and genes of the ergosterol biosynthesis pathway in Ku80R. Sterol analysis showed a significant decrease of the ergosterol mass under voriconazole exposure in Ku80, but not in Ku80R. However, the proportion of the sterol compounds was similar between both strains. To further assess the role of transporters, we used the ABC transporter inhibitor milbemycine oxime (MLB). MLB inhibited transporter activity in both Ku80 and Ku80R and demonstrated some potentiating effect on azole activity. Criteria for synergism were reached for MLB and posaconazole against Ku80. Finally, deletion of asg1 revealed some role of this transcription factor in controlling drug transporter expression, but had no impact on azole susceptibility. This work provides further insight in mechanisms of azole stress adaptation and suggests that drug transporters inhibition may represent a novel therapeutic target. Lay Summary A pan-azole-resistant strain was generated in vitro, in which drug transporter overexpression was a major trait. Analyses suggested a role of the transporter inhibitor milbemycin oxime in inhibiting drug transporters and potentiating azole activity.

Published
2021
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2. Emergence of azole resistant-Aspergillus fumigatus infections during STAT3-deficiency

Author

Marie-Elisabeth Bougnoux, Claire Givel, Fanny Lanternier, Florence Persat, Olivier Lortholary, Cécile Gautier, Dea Garcia-Hermoso, A. Duréault, Louis-Jean Couderc, Agathe Senechal, François Danion, Centre d'infectiologie Necker-Pasteur [CHU Necker], Institut Pasteur [Paris] (IP)-CHU Necker - Enfants Malades [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Aspergillus, Institut Pasteur [Paris] (IP), Imagine - Institut des maladies génétiques (IHU) (Imagine - U1163), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Centre National de Référence Mycoses Invasives et Antifongiques - National Reference Center Invasive Mycoses & Antifungals (CNRMA), Services de Pneumologie, Exploration Fonctionnelle Respiratoire et Cardiologie (Hôpital Louis Pradel), Hospices Civils de Lyon (HCL), Université de Lyon, Institut des Agents Infectieux [Lyon] (IAI), Unité de Parasitologie-Mycologie, Service de Microbiologie [Hôpital Necker-Enfants-Malades, Paris], Assistance Publique - Hôpitaux de Paris, Université Paris Cité (UPCité), Biologie et Pathogénicité fongiques - Fungal Biology and Pathogenicity (BPF), Institut Pasteur [Paris] (IP)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Service de pneumologie [Hôpital Foch], Hôpital Foch [Suresnes], UPRES EA220, Université de Versailles Saint-Quentin-en-Yvelines - UFR Sciences de la santé Simone Veil (UVSQ Santé), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), This work was supported by the French National Reference Center for Primary Immunodeficiencies (CEREDIH, Hôpital Necker-Enfants malades, Assistance Publique-Hôpitaux de Paris, Paris, France). F.D. and F.L. received a research grant from CEREDIH (Centre de Référence des Déficits Immunitaires Héréditaires, France)., Institut Pasteur [Paris]-CHU Necker - Enfants Malades [AP-HP], Institut Pasteur [Paris], Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Paris (UP), Université de Paris (UP), and Institut Pasteur [Paris]-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)

Subjects
Aspergillosis, MLP, Aspergillus fumigatus, STAT3, MESH: Genotype, chemistry.chemical_compound, CMC, Amphotericin B, MESH: Child, MESH: Azoles, microsatellite length polymorphism, aspergillosis, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, chemistry.chemical_classification, 0303 health sciences, MESH: Microbial Sensitivity Tests, biology, MESH: STAT3 Transcription Factor, General Medicine, STAT3-deficiency. Abbreviations: ABPA, CCPA, amphotericin B, 3. Good health, MESH: Caspofungin, MESH: Young Adult, signal transducer and activator of transcription 3, MESH: Communicable Diseases, MESH: Fungal Proteins, MESH: Pulmonary Aspergillosis, MESH: Aspergillus fumigatus, medicine.drug, Microbiology (medical), STAT3-deficiency, chronic cavitary pulmonary aspergillosis, azoles, chronic mucocutaneous candidiasis, Microbiology, MESH: Drug Resistance, Fungal, resistance, 03 medical and health sciences, hyper-IgE syndrome, MESH: Amphotericin B, medicine, allergic bronchopulmonary aspergillosis, 030304 developmental biology, invasive aspergillosis, MESH: Humans, 030306 microbiology, business.industry, IA, Retrospective cohort study, AmB, MESH: Adult, MESH: Retrospective Studies, biology.organism_classification, medicine.disease, MESH: Antifungal Agents, MESH: Male, MESH: France, chemistry, Primary immunodeficiency, Azole, Caspofungin, business
Abstract

Introduction. Signal transducer and activator of transcription 3 (STAT3) deficiency is a rare primary immunodeficiency associated with increased susceptibility to bacterial and fungal infections, notably pulmonary aspergillosis. Aim. We describe the emergence of azole-resistant Aspergillus fumigatus infections in STAT3-deficient patients. Methodology. During a retrospective study of 13 pulmonary aspergillosis cases in STAT3-deficient patients conducted in France, we identified patients infected with azole-resistant A. fumigatus isolates. Results. Two out of the 13 STAT3-deficient patients with aspergillosis had azole-resistant A. fumigatus infection, indicating an unexpectedly high prevalence of resistance. The first patient with STAT3 deficiency presented several flares of allergic bronchopulmonary aspergillosis-like episodes. He was chronically infected with two azole-resistant A. fumigatus isolates (TR34/L98). Despite prolonged antifungal treatment, including caspofungin and amphotericin B, the patient was not able to clear the azole-resistant A. fumigatus. The second patient had chronic cavitary pulmonary aspergillosis (CCPA). The A. fumigatus isolate was initially azole susceptible but harboured three F46Y, M172V and E427K point mutations. Despite prolonged antifungal therapies, lesions worsened and the isolate became resistant to all azoles. Surgery and caspofungin treatments were then required to cure CCPA. Resistance was probably acquired from the environment (TR34/L98) in the first case whereas resistance developed under antifungal treatments in the second case. These infections required long-term antifungal treatments and surgery. Conclusions. The emergence of azole-resistant A. fumigatus infections in STAT3-deficiency dramatically impacts both curative and prophylactic antifungal strategies. Physicians following patients with primary immune-deficiencies should be aware of this emerging problem as it complicates management of the patient.

Published
2020
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3. Short-term evolution strategies for host adaptation and drug escape in human fungal pathogens

Author

Chapman N. Beekman, Iuliana V. Ene, Brown University, and This work was supported by an NIH/NIAID R21AI139592 and an NIH/NIGMS IDeA award P20GM109035 to IVE.

Subjects
Antifungal Agents, [SDV]Life Sciences [q-bio], Yeast and Fungal Models, Drug resistance, Pathology and Laboratory Medicine, Pearls, Medicine and Health Sciences, Biology (General), Candida albicans, MESH: Evolution, Molecular, media_common, Candida, Genetics, Fungal Pathogens, 0303 health sciences, biology, Antimicrobials, Fungal genetics, Microevolution, Eukaryota, Drugs, Genomics, Adaptation, Physiological, 3. Good health, Experimental Organism Systems, Medical Microbiology, Host-Pathogen Interactions, Host adaptation, Pathogens, Drug, MESH: Fungi, Evolutionary Processes, QH301-705.5, media_common.quotation_subject, Immunology, Mycology, Research and Analysis Methods, Microbiology, MESH: Drug Resistance, Fungal, Evolution, Molecular, 03 medical and health sciences, MESH: Mycoses, Drug Resistance, Fungal, Virology, Microbial Control, Humans, Candida Albicans, Point Mutation, Fungal Genetics, Molecular Biology, Microbial Pathogens, Fungal Genomics, 030304 developmental biology, Pharmacology, Antifungals, Evolutionary Biology, MESH: Humans, 030306 microbiology, MESH: Host-Pathogen Interactions, Fungi, Organisms, Biology and Life Sciences, RC581-607, biology.organism_classification, MESH: Antifungal Agents, MESH: Adaptation, Physiological, Yeast, Mycoses, Mutation, Animal Studies, Parasitology, Fungal genome, Antimicrobial Resistance, Adaptation, Immunologic diseases. Allergy
Abstract

International audience; Research on human fungal pathogens has historically taken a backseat to other infectious diseases, perhaps due to a common misperception that fungi largely cause superficial infections [1]. In reality, fungi can be life-threatening to those who become immunocompromised during medical procedures or through conditions such as HIV and diabetes. Invasive fungal infections are estimated to kill over 1 million people every year, with mortality rates reaching 50% [2]. Significant challenges to the treatment of fungal infections include the limited availability of antifungals and the innate ability of fungi to rapidly evolve and adapt to fluctuating conditions. This adaptive ability is partially driven by extensive genomic plasticity, with many species acquiring diverse ploidy states, chromosomal rearrangements, and point mutations during host colonization [3-8]. Genetic plasticity enables rapid increases in virulence and antifungal drug resistance, which often translate to poor disease outcomes. Short-term evolution (microevolution) strategies in fungal pathogens are therefore essential for environmental adaptation in the mammalian host, and their study can inform adaptive mechanisms in other eukaryotes.

Published
2020
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4. Tracing the Evolutionary History and Global Expansion of Candida auris Using Population Genomic Analyses

Author

Ana Belén Araúz, Elizabeth L. Berkow, Oliver Kurzai, Christopher H. Heath, Rodney Adam, Sahar Althawadi, Xiao Li, Shawn R. Lockhart, Patricia Escandón, Ronen Ben-Ami, Anastasia P. Litvintseva, Lalitha Gade, Alexandre Alanio, Revathi Gunturu, Kaitlin Forsberg, Ana Alastruey-Izquierdo, Nancy A. Chow, Christina A. Cuomo, Amrita Bharat, Marie Desnos-Ollivier, Dianne Gardam, Belinda Calvo, Ronny Martin, José F. Muñoz, Rory M. Welsh, Centers for Disease Control and Prevention (CDC), Broad Institute [Cambridge], Massachusetts Institute of Technology (MIT)-Harvard University [Cambridge], Aga Khan University Hospital (AKUH), Nairobi, Mycologie moléculaire - Molecular Mycology, Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Centre National de Référence Mycoses Invasives et Antifongiques - National Reference Center Invasive Mycoses & Antifungals (CNRMA), Institut Pasteur [Paris], Laboratoire de Parasitologie-Mycologie [CHU Saint Louis, Paris], Groupe Hospitalier Saint Louis - Lariboisière - Fernand Widal [Paris], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Université de Paris (UP), Instituto de Salud Carlos III [Madrid] (ISC), King Faisal Specialist Hospital and Research Centre, Riyadh, Hospital Santo Tomás, Tel Aviv Sourasky Medical Center [Te Aviv], Tel Aviv University [Tel Aviv], Public Health Agency of Canada (PHAC), Universidad del Zulia (LUZ), Instituto Nacional de Salud [Bogota], Fiona Stanley Hospital [Murdoch], Royal Perth Hospital, The University of Western Australia (UWA), Leibniz Institute for Natural Product Research and Infection Biology (Hans Knoell Institute), University of Würzburg = Universität Würzburg, This project has been funded in part with federal funds from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services, under award U19AI110818 to the Broad Institute. C.A.C. is a CIFAR fellow in the Fungal Kingdom Program. This work was also made possible through support from the Advanced Molecular Detection (AMD) initiative at CDC., National Institute of Allergy and Infectious Diseases (United States), Harvard University-Massachusetts Institute of Technology (MIT), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP), Université Paris Cité (UPCité), King Faisal Specialist Hospital and Research Centre (KFSH & RC), and Tel Aviv University (TAU)

Subjects
Azoles, Antifungal Agents, Population genetics, Emerging species, Echinocandins, MESH: Azoles, Molecular clock, Clade, MESH: Phylogeny, Fluconazole, Phylogeny, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, Candida, genome analysis, emerging species, 0303 health sciences, education.field_of_study, Molecular Epidemiology, Candida auris, antifungal resistance, Biological Evolution, QR1-502, 3. Good health, Phylogeography, MESH: Phylogeography, MESH: Genome, Fungal, MESH: Fluconazole, Genome, Fungal, MESH: Metagenomics, MESH: Whole Genome Sequencing, Research Article, MESH: Mutation, Population, Genes, Fungal, MESH: Genetics, Population, Context (language use), MESH: Biological Evolution, Ecological and Evolutionary Science, Biology, Microbiology, MESH: Drug Resistance, Fungal, 03 medical and health sciences, Phylogenetics, Drug Resistance, Fungal, MESH: Candida, Virology, Humans, MESH: Molecular Epidemiology, Candidiasis, Invasive, education, 030304 developmental biology, MESH: Humans, Molecular epidemiology, Whole Genome Sequencing, 030306 microbiology, MESH: Echinocandins, population genetics, Antifungal resistance, Genome analysis, MESH: Antifungal Agents, MESH: Candidiasis, Invasive, Genetics, Population, Evolutionary biology, Mutation, Metagenomics, MESH: Genes, Fungal
Abstract

In less than a decade, C. auris has emerged in health care settings worldwide; this species is capable of colonizing skin and causing outbreaks of invasive candidiasis. In contrast to other Candida species, C. auris is unique in its ability to spread via nosocomial transmission and its high rates of drug resistance. As part of the public health response, whole-genome sequencing has played a major role in characterizing transmission dynamics and detecting new C. auris introductions. Through a global collaboration, we assessed genome evolution of isolates of C. auris from 19 countries. Here, we described estimated timing of the expansion of each C. auris clade and of fluconazole resistance, characterized discrete phylogeographic population structure of each clade, and compared genome data to sensitivity measurements to describe how antifungal resistance mechanisms vary across the population. These efforts are critical for a sustained, robust public health response that effectively utilizes molecular epidemiology., Candida auris has emerged globally as a multidrug-resistant yeast that can spread via nosocomial transmission. An initial phylogenetic study of isolates from Japan, India, Pakistan, South Africa, and Venezuela revealed four populations (clades I, II, III, and IV) corresponding to these geographic regions. Since this description, C. auris has been reported in more than 30 additional countries. To trace this global emergence, we compared the genomes of 304 C. auris isolates from 19 countries on six continents. We found that four predominant clades persist across wide geographic locations. We observed phylogeographic mixing in most clades; clade IV, with isolates mainly from South America, demonstrated the strongest phylogeographic substructure. C. auris isolates from two clades with opposite mating types were detected contemporaneously in a single health care facility in Kenya. We estimated a Bayesian molecular clock phylogeny and dated the origin of each clade within the last 360 years; outbreak-causing clusters from clades I, III, and IV originated 36 to 38 years ago. We observed high rates of antifungal resistance in clade I, including four isolates resistant to all three major classes of antifungals. Mutations that contribute to resistance varied between the clades, with Y132F in ERG11 as the most widespread mutation associated with azole resistance and S639P in FKS1 for echinocandin resistance. Copy number variants in ERG11 predominantly appeared in clade III and were associated with fluconazole resistance. These results provide a global context for the phylogeography, population structure, and mechanisms associated with antifungal resistance in C. auris.

Published
2020
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5. Tunisian population of Mycosphaerella graminicola is still sensitive to strobilurin fungicides

Author

M, Naouari, A, Siah, M, Elgazzah, Ph, Reignault, P, Halama, Prévot-Liger, Perrine, Unité de Chimie Environnementale et Interactions sur le Vivant (UCEIV), and Université du Littoral Côte d'Opale (ULCO)

Subjects
Tunisia, [SDV]Life Sciences [q-bio], food and beverages, MESH: Triticum, MESH: Ascomycota, MESH: Fungicides, Industrial, Fungicides, Industrial, MESH: Drug Resistance, Fungal, [SDV] Life Sciences [q-bio], MESH: Plant Diseases, Ascomycota, Drug Resistance, Fungal, MESH: Tunisia, Triticum, Plant Diseases
Abstract

International audience; Septoria tritici blotch caused by the fungal pathogen Mycosphaerella graminicola (anamorph: Zymoseptoria tritici) is one of the most frequently occurring diseases on both bread and durum wheat crops worldwide. One hundred and sixty four durum wheat-adapted isolates of this fungus were sampled during the 2012 growing season from five distinct geographical locations of Tunisia (Bizerte, Béja, Kef, Jendouba and Siliana) in order to examine the status of strobilurin resistance of M. graminicola in this country. Resistance was assessed by screening the G143A substitution (Cytochrome b) which confers resistance to this class of fungicides. We used a PCR-based mismatch mutation assay allowing the amplification of either G143 (sensitive) or A143 (resistant) allele. All isolates were found to contain the sensitive wild-type G143 allele and therefore to be sensitive. Our study confirms recent reports on M. graminicola in Tunisia and shows that the Tunisian population of the fungus remains fully sensitive to strobilurins. A durability-oriented management of strobilurin applications in Tunisia is thereby recommended to prevent the development and widespread of the corresponding resistance such as in Europe, where pathogen populations are nowadays fully resistant to strobilurins.

Published
2020

6. Predisposing factors and outcome of uncommon yeast species-related fungaemia based on an exhaustive surveillance programme (2002–14)

Author

Charlotte Renaudat, Stéphane Bretagne, Olivier Lortholary, Françoise Dromer, Karine Sitbon, Marie Desnos-Ollivier, Service de parasitologie [Mondor], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Mycologie moléculaire, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Centre National de Référence Mycoses Invasives et Antifongiques - National Reference Center Invasive Mycoses & Antifungals (CNRMA), Institut Pasteur [Paris], Université Paris Descartes - Paris 5 (UPD5), The YEASTS programme was supported in part by Institut de Veille Sanitaire and Institut Pasteur., The authors thank all the physicians and technicians in the 27 hospitals who made this study possible, The following investigators participated in the YEASTS programme of the French Mycosis Study Group. For data collection in each participating centre: Claire Bouges-Michel, Sophie Brun (hôpital Avicenne, Bobigny), Isabelle Poilane (hôpital Jean Verdier, Bondy), Jean Dunand, Nawel Ait Ammar (hôpital Ambroise Paré, Boulogne), Guy Galeazzi (hôpital Louis Mourier, Colombes), Stéphane Bretagne, Françoise Botterel, Françoise Foulet, Nawel Ait Ammar (hôpital Henri Mondor, Créteil), Nathalie Fauchet (Centre Intercommunal, Créteil), Elisabeth Forget (hôpital Beaujon, Clichy), Christine Lawrence, Anne-Laure Roux (hôpital Raymond Poincaré, Garches), Adela Enache-Angoulvant, Christine Bonnal, Françoise Botterel (hôpital du Kremlin Bicêtre, Kremlin-Bicêtre), Odile Eloy (Centre Hospitalier, Le Chesnay), Marie-France David, Najiby Kassis, Lilia Mihaila (hôpital Paul Brousse, Villejuif), Elisabeth Chachaty (Institut Gustave Roussy, Villejuif), and in Paris: Christian Chochillon (hôpital Bichat), André Paugam, Marie-Thérèse Baixench, Florence Lesle, Nada Kherbeck-Hassoun (hôpital Cochin), Marie-Christine Escande (Institut Curie), Marie-Elisabeth Bougnoux, Yvon Sterckers, Svetlana Challier (hôpital Necker), Eric Dannaoui, Véronique Lavarde (hôpital Européen Georges Pompidou), Annick Datry, Bader Lmimouni, Sophie Brun, Arnaud Fekkar (hôpital de la Pitié-Salpétrière), Jean-Louis Poirot, Juliette Guitard (hôpital Saint Antoine), Alexandre Alanio, Stéphane Bretagne, Claire Lacroix (hôpital Saint Louis), Didier Moissenet (hôpital Trousseau), Muriel Cornet (Hôtel Dieu), Michel Develoux (hôpital Tenon), Stéphane Bonacorsi, Patricia Mariani, Catherine Doit (hôpital Robert Debré), and for mycological expertise at the CNRMA: Catherine Blanc, Anne Boullié, Dea Garcia-Hermoso, Damien Hoinard and Dorothée Raoux-Barbot. Funding, Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris] (IP)

Subjects
0301 basic medicine, Male, Antifungal Agents, Candida parapsilosis, Candida tropicalis, MESH: Aged, 80 and over, Candida albicans, Pharmacology (medical), Prospective Studies, MESH: Phylogeny, Fungemia, Phylogeny, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, Aged, 80 and over, MESH: Aged, MESH: Microbial Sensitivity Tests, MESH: Middle Aged, biology, Candidiasis, Middle Aged, MESH: Candidiasis, 3. Good health, Causality, Infectious Diseases, MESH: Young Adult, Epidemiological Monitoring, Female, France, medicine.drug, Microbiology (medical), Adult, Adolescent, 030106 microbiology, Microbial Sensitivity Tests, MESH: Ascomycota, MESH: Causality, Microbiology, MESH: Drug Resistance, Fungal, 03 medical and health sciences, Young Adult, Ascomycota, Drug Resistance, Fungal, MESH: Fungemia, Candida krusei, Trichosporon, medicine, Humans, Aged, Pharmacology, MESH: Adolescent, MESH: Humans, Candida glabrata, business.industry, Basidiomycota, MESH: Candida albicans, MESH: Adult, biology.organism_classification, medicine.disease, MESH: Antifungal Agents, MESH: Male, MESH: Prospective Studies, MESH: France, MESH: Basidiomycota, MESH: Epidemiological Monitoring, business, MESH: Female, Fluconazole
Abstract

International audience; Objectives:Using registry data to compare fungaemia caused by uncommon yeast species (UYS; i.e. other than Candida albicans , Candida glabrata , Candida parapsilosis , Candida tropicalis and Candida krusei ) and C. albicans -related fungaemia can reveal specific predisposing factors of UYS with potential impact on treatment strategies.Methods:We analysed 338 episodes of UYS fungaemia prospectively collected from 27 hospitals (Paris, France; 1 October 2002-31 December 2014) and compared these with 1998 single episodes of C. albicans fungaemia using univariate and multivariate analyses.Results:The proportion of UYS fungaemia was stable over time. Thirty-five different species were identified (27 ascomycetes, 8 basidiomycetes), 11 had caspofungin MIC 50 >0.25 mg/L and 15 fluconazole MIC 50 >4 mg/L. Haematological malignancies [OR=2.39 (95% CI 1.79-3.18)] and prior exposure to antifungal drugs [OR=1.87 (1.30-2.69)] were independent predisposing factors for UYS infections upon multivariate analysis. However, when considering the genus/species complex level, only infections due to Candida kefyr -related species [OR=4.01 (2.42-6.64)] and to Trichosporon spp. [OR=5.38 (1.72-16.81)] remained associated with haematological malignancies, those due to the GEOTRICHUM group with acute leukaemia [OR=61.29 (19.23-195.36)], and infections with Trichosporon spp. or the GEOTRICHUM group with prior exposure to caspofungin [OR=15.67 (3.62-67.80) and OR=13.17 (3.33-52.03), respectively] but not to fluconazole. The global mortality at day 30 for UYS was similar to that for C. albicans (35.4%, and 39.9%, respectively), but very divergent results were observed according to the specific UYS.Conclusions:UYS encompass a high diversity of species, each with its own behaviour and predisposing factors for human infections. This variety makes it important to rapidly identify an isolate to the species level in order to optimize antifungal treatment.

Published
2017
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7. Cell Wall Changes in Amphotericin B-Resistant Strains from Candida tropicalis and Relationship with the Immune Responses Elicited by the Host

Author

Jesús Pla, Ana Cecilia Mesa-Arango, Mihai G. Netea, Elvira Román, Cristina Rueda, Jessica Quintin, Oscar Zaragoza, María C. Terrón, Daniel Luque, Instituto de Salud Carlos III [Madrid] (ISC), Universidad de Antioquia = University of Antioquia [Medellín, Colombia], Universidad Complutense de Madrid = Complutense University of Madrid [Madrid] (UCM), Radboud University Medical Center [Nijmegen], and A. C. Mesa-Arango has been supported by fellowships from Fundación Carolina and Instituto de Salud Carlos III. C. Rueda has a Sara Borrell contract from the Fondo de Investigaciones Sanitarias (reference no. CD11/00110).

Subjects
0301 basic medicine, Antifungal Agents, beta-Glucans, [SDV]Life Sciences [q-bio], lnfectious Diseases and Global Health Radboud Institute for Molecular Life Sciences [Radboudumc 4], Chitin, Drug resistance, Moths, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, MESH: Candida tropicalis, MESH: Chitin, Candida tropicalis, MESH: Melanins, Cell Wall, Amphotericin B, Pharmacology (medical), MESH: Animals, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, MESH: Microbial Sensitivity Tests, MESH: beta-Glucans, Kinase, MESH: Moths, 3. Good health, MESH: Leukocytes, Mononuclear, Infectious Diseases, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Larva, Host-Pathogen Interactions, [SDV.IMM]Life Sciences [q-bio]/Immunology, MESH: Immunity, Innate, Mitogen-Activated Protein Kinases, medicine.drug, 030106 microbiology, Microbial Sensitivity Tests, Biology, Microbiology, Proinflammatory cytokine, MESH: Drug Resistance, Fungal, Cell wall, 03 medical and health sciences, Immune system, MESH: Cell Wall, Drug Resistance, Fungal, Mechanisms of Resistance, Immunity, MESH: Amphotericin B, medicine, Animals, Humans, Melanins, Pharmacology, MESH: Humans, MESH: Host-Pathogen Interactions, Congo Red, biology.organism_classification, MESH: Antifungal Agents, MESH: Mitogen-Activated Protein Kinases, Immunity, Innate, MESH: Congo Red, Leukocytes, Mononuclear, MESH: Larva
Abstract

We have morphologically characterized Candida tropicalis isolates resistant to amphotericin B (AmB). These isolates present an enlarged cell wall compared to isolates of regular susceptibility. This correlated with higher levels of β-1,3-glucan in the cell wall but not with detectable changes in chitin content. In line with this, AmB-resistant strains showed reduced susceptibility to Congo red. Moreover, mitogen-activated protein kinases (MAPKs) involved in cell integrity were already activated during regular growth in these strains. Finally, we investigated the response elicited by human blood cells and found that AmB-resistant strains induced a stronger proinflammatory response than susceptible strains. In agreement, AmB-resistant strains also induced stronger melanization of Galleria mellonella larvae, indicating that the effect of alterations of the cell wall on the immune response is conserved in different types of hosts. Our results suggest that resistance to AmB is associated with pleiotropic mechanisms that might have important consequences, not only for the efficacy of the treatment but also for the immune response elicited by the host.

Published
2016
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8. High prevalence of triazole resistance in Aspergillus fumigatus, especially mediated by TR/L98H, in a French cohort of patients with cystic fibrosis

Author

Emilie Sacchetto, Alain Haloun, Guillaume Ghislain Aubin, Patrice Le Pape, Dea Garcia-Hermoso, Isabelle Danner-Boucher, Florent Morio, Michel Miegeville, Stéphane Bretagne, Université de Nantes (UN), Centre hospitalier universitaire de Nantes (CHU Nantes), Centre National de Référence des Mycoses invasives et antifongiques - Mycologie moléculaire (CNRMA), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), This study was supported by internal funding., Cibles et Médicaments des Infections et de l'Immunité (IICiMed), Université de Nantes - UFR des Sciences Pharmaceutiques et Biologiques, Université de Nantes (UN)-Université de Nantes (UN), and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Male, MESH: Sequence Analysis, DNA, Posaconazole, Antifungal Agents, Cystic Fibrosis, MESH: Tubulin, Cystic fibrosis, Aspergillus fumigatus, Tubulin, MESH: Child, Prevalence, Agar, Pharmacology (medical), CFTR, Child, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, MESH: Microbial Sensitivity Tests, 0303 health sciences, multi-azole resistance, MESH: Middle Aged, biology, cyp51A mutations, MESH: Voriconazole, Middle Aged, 3. Good health, Infectious Diseases, MESH: Young Adult, Child, Preschool, France, Itraconazole, MESH: Aspergillus fumigatus, medicine.drug, Adult, Microbiology (medical), food.ingredient, Adolescent, MESH: Cystic Fibrosis, Microbial Sensitivity Tests, In Vitro Techniques, A. fumigatus, MESH: Drug Resistance, Fungal, Microbiology, Agar plate, Young Adult, 03 medical and health sciences, food, Drug Resistance, Fungal, medicine, Aspergillosis, Humans, MESH: Aspergillosis, Gene, MESH: Prevalence, Retrospective Studies, 030304 developmental biology, MESH: Adolescent, MESH: In Vitro Techniques, Pharmacology, Voriconazole, MESH: Humans, 030306 microbiology, MESH: Child, Preschool, MESH: Adult, MESH: Retrospective Studies, Sequence Analysis, DNA, Triazoles, MESH: Antifungal Agents, medicine.disease, biology.organism_classification, MESH: Male, MESH: France, Pyrimidines, MESH: Triazoles, MESH: Pyrimidines, MESH: Itraconazole
Abstract

Part of this work has been presented at the Fifth Congress on Trends in Medical Mycology, Valencia, Spain, 2011 (Abstract no. 308); International audience; OBJECTIVES:Triazole resistance in Aspergillus fumigatus due to a single azole resistance mechanism (TR/L98H) is increasingly reported in European countries. Data from patients with cystic fibrosis (CF) are limited. Our study aimed to investigate the prevalence and molecular mechanisms of azole resistance in A. fumigatus in a cohort of patients with CF.METHODS:Eighty-five A. fumigatus isolates from 50 CF patients, collected between January 2010 and April 2011, were retrospectively analysed for azole resistance using agar plates containing 4 mg/L itraconazole. MICs of itraconazole, voriconazole and posaconazole were determined according to EUCAST methodology for each isolate able to grow on this medium. Species identification was performed by sequencing of the β-tubulin gene. Sequencing analysis of the cyp51A gene and its promoter region was conducted.RESULTS:Nine isolates (four patients, 8% prevalence) were able to grow on itraconazole-containing agar plates. Itraconazole resistance was confirmed by EUCAST methodology (MICs >2 mg/L). All isolates had mutations in the cyp51A gene at residues previously involved in azole resistance: L98H (n = 5), M220T (n = 4) and G54R (n = 1). One patient had three genetically distinct azole-resistant isolates identified during the study. The isolates with L98H that were recovered from three patients (6% prevalence) also had the 34 bp tandem repeat in the promoter region of cyp51A (TR/L98H) and displayed multiazole resistance.CONCLUSIONS:We report an 8% prevalence of itraconazole resistance in CF patients in our centre, mostly driven by TR/L98H (6%). Our data confirm that TR/L98H occurs in France and can be highly prevalent in CF patients.

Published
2012
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9. Prior Caspofungin Exposure in Patients with Hematological Malignancies Is a Risk Factor for Subsequent Fungemia Due to Decreased Susceptibility in Candida spp.: a Case-Control Study in Paris, France

Author

Karine Boukris-Sitbon, Olivier Lortholary, Marie Desnos-Ollivier, Françoise Dromer, Didier Guillemot, Elodie Blanchard, Centre de Référence Déficits Immunitaires Héréditaires (CEREDIH), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Necker - Enfants Malades [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Centre d'infectiologie Necker-Pasteur [CHU Necker], Institut Pasteur [Paris]-CHU Necker - Enfants Malades [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Mycologie moléculaire, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Centre National de Référence Mycologie et Antifongiques-Mycologie Moléculaire (CNRMA), Institut Pasteur [Paris], Thérapeutique Recombinante Expérimentale (TIMC-IMAG-TheREx), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Centre de Référence Déficits Immunitaires Héréditaires ( CEREDIH ), Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Necker - Enfants Malades [AP-HP], Institut Pasteur [Paris]-Centre National de la Recherche Scientifique ( CNRS ), Centre National de Référence Mycologie et Antifongiques-Mycologie Moléculaire ( CNRMA ), Thérapeutique Recombinante Expérimentale ( TIMC-IMAG-THEREX ), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications [Grenoble] ( TIMC-IMAG ), Université Joseph Fourier - Grenoble 1 ( UJF ) -Institut polytechnique de Grenoble - Grenoble Institute of Technology ( Grenoble INP ) -IMAG-Centre National de la Recherche Scientifique ( CNRS ) -Université Grenoble Alpes ( UGA ) -Université Joseph Fourier - Grenoble 1 ( UJF ) -Institut polytechnique de Grenoble - Grenoble Institute of Technology ( Grenoble INP ) -IMAG-Centre National de la Recherche Scientifique ( CNRS ) -Université Grenoble Alpes ( UGA ), Institut Pasteur [Paris] (IP)-CHU Necker - Enfants Malades [AP-HP], Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP), and Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Male, Antifungal Agents, Multivariate analysis, MESH : Aged, Drug resistance, Echinocandins, chemistry.chemical_compound, 0302 clinical medicine, Caspofungin, Risk Factors, MESH: Risk Factors, polycyclic compounds, MESH : Female, Pharmacology (medical), 030212 general & internal medicine, [ SDV.MP.MYC ] Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, Fungemia, Candida, MESH: Aged, 0303 health sciences, MESH: Middle Aged, Middle Aged, MESH : Risk Factors, MESH: Case-Control Studies, 3. Good health, Infectious Diseases, Hematologic Neoplasms, Female, France, MESH : Hematologic Neoplasms, MESH : Case-Control Studies, medicine.medical_specialty, MESH : Male, MESH : Antifungal Agents, Clinical Therapeutics, Biology, MESH: Drug Resistance, Fungal, Lipopeptides, 03 medical and health sciences, Drug Resistance, Fungal, MESH: Fungemia, MESH: Candida, Internal medicine, medicine, Humans, MESH : Middle Aged, Risk factor, MESH : France, Aged, MESH : Echinocandins, Pharmacology, MESH: Humans, 030306 microbiology, MESH: Echinocandins, MESH : Humans, Case-control study, MESH : Drug Resistance, Fungal, Odds ratio, bacterial infections and mycoses, MESH: Antifungal Agents, medicine.disease, MESH: Male, Confidence interval, MESH: France, MESH : Fungemia, chemistry, Case-Control Studies, Immunology, MESH: Female, MESH : Candida, MESH: Hematologic Neoplasms
Abstract

Infections due to caspofungin-resistant Candida isolates in patients exposed to caspofungin therapy are increasing. We report here a nested case-control study which aimed at identifying factors associated with bloodstream infections caused by Candida spp. having reduced susceptibility to caspofungin (CRSC) in adults suffering from hematological malignancies. In univariate and multivariate analyses, infections with CRSC were associated with caspofungin exposure in the previous 30 days (odds ratio [OR] = 5.25; 95% confidence interval [95% CI], 1.68–16.35) and with an age of ≤65 years (OR = 3.27; 95% CI, 1.26–8.50).

Published
2011
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10. Interaction of Candida albicans Biofilms with Antifungals: Transcriptional Response and Binding of Antifungals to Beta-Glucans

Author

Christophe d'Enfert, Tristan Rossignol, Govindsamy Vediyappan, Biologie et Pathogénicité fongiques (BPF), Institut National de la Recherche Agronomique (INRA)-Institut Pasteur [Paris] (IP), We are grateful to Caroline Proux and Jean-Yves Coppé at the PF2 Platform of Pasteur Genopole Ile-de-France for help with microarray experiments. Caspofungin was kindly provided by Merck & Co. Fluconazole was kindly provided by Pfizer Inc.This work was supported by a grant from the European Commission to C.D. (EURESFUN, LSHM-CT-2005-518199). G.V. was the recipient of a postdoctoral fellowship in the framework of the EURESFUN Consortium. T.R. was the recipient of a postdoctoral fellowship in the framework of the NPARI Consortium (LSHE-CT-2006-037692)., Biologie et Pathogénicité fongiques, Institut National de la Recherche Agronomique (INRA)-Institut Pasteur [Paris], and Institut Pasteur [Paris]-Institut National de la Recherche Agronomique (INRA)

Subjects
MESH: Extracellular Matrix Proteins, Antifungal Agents, beta-Glucans, Transcription, Genetic, MESH: Membrane Glycoproteins, Extracellular matrix, Echinocandins, chemistry.chemical_compound, MESH: Lipopeptides, Caspofungin, Gene Expression Regulation, Fungal, Candida albicans, Gene expression, Pharmacology (medical), Luciferases, Fluconazole, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, Extracellular Matrix Proteins, 0303 health sciences, Membrane Glycoproteins, MESH: beta-Glucans, Corpus albicans, Infectious Diseases, MESH: Fluconazole, MESH: Fungal Proteins, MESH: Gene Expression Regulation, Fungal, medicine.drug, MESH: Biofilms, Biology, MESH: Drug Resistance, Fungal, Microbiology, Fungal Proteins, MESH: Gene Expression Profiling, Lipopeptides, 03 medical and health sciences, Drug Resistance, Fungal, Amphotericin B, MESH: Amphotericin B, medicine, Mechanisms of Action: Physiological Effects, 030304 developmental biology, Pharmacology, 030306 microbiology, MESH: Transcription, Genetic, MESH: Candida albicans, MESH: Echinocandins, Gene Expression Profiling, Biofilm, biochemical phenomena, metabolism, and nutrition, MESH: Antifungal Agents, biology.organism_classification, In vitro, chemistry, Biofilms, MESH: Luciferases
Abstract

Candida albicans can form biofilms that exhibit elevated intrinsic resistance to various antifungal agents, in particular azoles and polyenes. The molecular mechanisms involved in the antifungal resistance of biofilms remain poorly understood. We have used transcript profiling to explore the early transcriptional responses of mature C. albicans biofilms exposed to various antifungal agents. Mature C. albicans biofilms grown under continuous flow were exposed for as long as 2 h to concentrations of fluconazole (FLU), amphotericin B (AMB), and caspofungin (CAS) that, while lethal for planktonic cells, were not lethal for biofilms. Interestingly, FLU-exposed biofilms showed no significant changes in gene expression over the course of the experiment. In AMB-exposed biofilms, 2.7% of the genes showed altered expression, while in CAS-exposed biofilms, 13.0% of the genes had their expression modified. In particular, exposure to CAS resulted in the upregulation of hypha-specific genes known to play a role in biofilm formation, such as ALS3 and HWP1 . There was little overlap between AMB- or CAS-responsive genes in biofilms and those that have been identified as AMB, FLU, or CAS responsive in C. albicans planktonic cultures. These results suggested that the resistance of C. albicans biofilms to azoles or polyenes was due not to the activation of specific mechanisms in response to exposure to these antifungals but rather to the intrinsic properties of the mature biofilms. In this regard, our study led us to observe that AMB physically bound C. albicans biofilms and beta-glucans, which have been proposed to be major constituents of the biofilm extracellular matrix and to prevent azoles from reaching biofilm cells. Thus, enhanced extracellular matrix or beta-glucan synthesis during biofilm growth might prevent antifungals, such as azoles and polyenes, from reaching biofilm cells, thus limiting their toxicity to these cells and the associated transcriptional responses.

Published
2010
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11. Functional characterization of the Saccharomyces cerevisiae ABC-transporter Yor1p overexpressed in plasma membranes

Author

Sylvain Blanquet, Myriam Lazard, Pierre Plateau, Ioana Grigoras, Laboratoire de Biochimie de l'Ecole polytechnique (BIOC), and École polytechnique (X)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Plasma membranes, Oligomycin, Antifungal Agents, Saccharomyces cerevisiae Proteins, ATPase, Saccharomyces cerevisiae, Biophysics, ATP-binding cassette transporter, ATPase activity, Biochemistry, MESH: Drug Resistance, Fungal, Substrate Specificity, chemistry.chemical_compound, MESH: Saccharomyces cerevisiae Proteins, ATP hydrolysis, Drug Resistance, Fungal, MESH: Adenosine Triphosphatases, Vanadate, Enzyme Inhibitors, Adenosine Triphosphatases, biology, Cell Membrane, Cell Biology, biology.organism_classification, MESH: Antifungal Agents, MESH: Saccharomyces cerevisiae, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], chemistry, Cyclic nucleotide-binding domain, MESH: Enzyme Inhibitors, biology.protein, MESH: ATP-Binding Cassette Transporters, MESH: Substrate Specificity, ATP-Binding Cassette Transporters, Electrophoresis, Polyacrylamide Gel, Yor1p, ABC transporter, Ethidium bromide, MESH: Cell Membrane, MESH: Electrophoresis, Polyacrylamide Gel
Abstract

International audience; Yor1p, a Saccharomyces cerevisiae plasma membrane ABC-transporter, is associated to oligomycin resistance and to rhodamine B transport. Here, by using the overexpressing strain Superyor [A. Decottignies, A.M. Grant, J.W. Nichols, H. de Wet, D.B. McIntosh, A. Goffeau, ATPase and multidrug transport activities of the overexpressed yeast ABC protein Yor1p, J. Biol. Chem. 273 (1998) 12612-12622], we show that Yor1p also confers resistance to rhodamine 6G and to doxorubicin. In addition, Yor1p protects cells, although weakly, against tetracycline, verapamil, eosin Y and ethidium bromide. The basal ATPase activity of the overexpressed form of Yor1p was studied in membrane preparations. This activity is quenched upon addition of micromolar amounts of vanadate. Vmax and Km values of approximately 0.8 s(-1) and 50+/-8 microM are measured. Mutations of essential residues in the nucleotide binding domain 2 reduces the activity to that measured with a Deltayor1 strain. ATP hydrolysis is strongly inhibited by the addition of potential substrates of the transporter. Covalent reaction of 8-azido-[alpha-(32)P]ATP with Yor1p is not sensitive to the presence of excess oligomycin. Thus, competition of the drug with ATP binding is unlikely. Finally, we inspect possible hypotheses accounting for substrate inhibition, rather than stimulation, of ATP hydrolysis by the membrane preparation.

Published
2008
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12. Next-generation sequencing offers new insights into the resistance of Candida spp. to echinocandins and azoles

Author

Danièle Maubon, Christophe Hennequin, Cécile Garnaud, Marie-Elisabeth Bougnoux, Natacha Sertour, Françoise Botterel, Sylvie Larrat, Muriel Cornet, Jesús Guinea, Eric Dannaoui, Thérapeutique Recombinante Expérimentale (TIMC-IMAG-TheREx), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Unité de Parasitologie-Mycologie [CHU HEGP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Européen Georges Pompidou [APHP] (HEGP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO), Biologie et Pathogénicité fongiques, Institut Pasteur [Paris]-Institut National de la Recherche Agronomique (INRA), CHU Necker - Enfants Malades [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Dynamic Microbiology - EA 7380 (DYNAMIC), École nationale vétérinaire d'Alfort (ENVA)-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Biologie structurale des interactions entre virus et cellule hôte (UVHCI), Université Joseph Fourier - Grenoble 1 (UJF)-European Molecular Biology Laboratory [Grenoble] (EMBL)-Centre National de la Recherche Scientifique (CNRS), Institut de Biologie et Pathologie [CHU Grenoble] (IBP), CHU Grenoble-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Centre d'Immunologie et de Maladies Infectieuses (CIMI), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC), Service de Parasitologie - Mycologie [CHU Saint-Antoine], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Saint-Antoine [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU), Clinical Microbiology and Infectious Diseases Department, Universidad Complutense de Madrid = Complutense University of Madrid [Madrid] (UCM), This work was supported by a grant from Pfizer (Prix de recherche Société Française de Mycologie Médicale—Pfizer 2012). The funder had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript., Thérapeutique Recombinante Expérimentale ( TIMC-IMAG-THEREX ), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications [Grenoble] ( TIMC-IMAG ), Université Joseph Fourier - Grenoble 1 ( UJF ) -Institut polytechnique de Grenoble - Grenoble Institute of Technology ( Grenoble INP ) -IMAG-Centre National de la Recherche Scientifique ( CNRS ) -Université Grenoble Alpes ( UGA ) -Université Joseph Fourier - Grenoble 1 ( UJF ) -Institut polytechnique de Grenoble - Grenoble Institute of Technology ( Grenoble INP ) -IMAG-Centre National de la Recherche Scientifique ( CNRS ) -Université Grenoble Alpes ( UGA ), Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Européen Georges Pompidou [APHP] ( HEGP ), Institut National de la Recherche Agronomique ( INRA ) -Institut Pasteur [Paris], Dynamyc Mycology - EA 7380, Université Paris-Est Créteil Val-de-Marne - Paris 12 ( UPEC UP12 ), Laboratoire de biologie structurale des interactions entre virus et cellule hôte, Université Joseph Fourier - Grenoble 1 ( UJF ) -Centre National de la Recherche Scientifique ( CNRS ), Institut de Biologie et Pathologie [CHU Grenoble] ( IBP ), CHU Grenoble-Université Grenoble Alpes ( UGA ), Centre d'Immunologie et de Maladies Infectieuses ( CIMI ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Saint-Antoine [APHP], Universidad Complutense de Madrid [Madrid] ( UCM ), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS), Biologie et Pathogénicité fongiques (BPF), Institut National de la Recherche Agronomique (INRA)-Institut Pasteur [Paris] (IP), École nationale vétérinaire - Alfort (ENVA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Université Paris-Est (UPE)-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), CHU Saint-Antoine [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP)-IMAG-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP)-IMAG-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), Hôpital Européen Georges Pompidou [APHP] (HEGP), Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP), Institut National de la Recherche Agronomique (INRA)-Institut Pasteur [Paris], Dynamyc- Dynamyc Mycology - EA 7380, Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), CHU Grenoble-Université Grenoble Alpes (UGA), Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-CHU Saint-Antoine [APHP], Universidad Complutense de Madrid [Madrid] (UCM), and Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)

Subjects
Azoles, Antifungal Agents, MESH : Genotype, Candida parapsilosis, MESH: Genotype, chemistry.chemical_compound, MESH : Candidiasis, Echinocandins, MESH: Azoles, Pharmacology (medical), Candida albicans, MESH: High-Throughput Nucleotide Sequencing, [ SDV.MP.MYC ] Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, Candida, Genetics, chemistry.chemical_classification, 0303 health sciences, biology, Candidiasis, High-Throughput Nucleotide Sequencing, Caspofungin Acetate, MESH: Candidiasis, Infectious Diseases, NGS, MESH : Mutation, antifungals, medicine.drug, Microbiology (medical), MESH: Mutation, Echinocandin, Genotype, MESH : Antifungal Agents, Microbiology, MESH: Drug Resistance, Fungal, 03 medical and health sciences, Drug Resistance, Fungal, MESH: Candida, medicine, Humans, 030304 developmental biology, MESH : Echinocandins, Pharmacology, MESH: Humans, Candida glabrata, 030306 microbiology, MESH: Echinocandins, MESH : Humans, MESH : Drug Resistance, Fungal, biology.organism_classification, mutations, MESH: Antifungal Agents, MESH : Azoles, chemistry, Mutation, Azole, Caspofungin, MESH : Candida, MESH : High-Throughput Nucleotide Sequencing
Abstract

International audience; ObjectivesMDR Candida strains are emerging. Next-generation sequencing (NGS), which enables extensive and deep genome analysis, was used to investigate echinocandin and azole resistance in clinical Candida isolates.; MethodsSix genes commonly involved in antifungal resistance (ERG11, ERG3, TAC1, CgPDR1, FKS1 and FKS2) were analysed using NGS in 40 Candida isolates (18 Candida albicans, 15 Candida glabrata and 7 Candida parapsilosis). The strategy was validated using strains with known sequences. Then, 8 clinical strains displaying antifungal resistance and 23 sequential isolates collected from 10 patients receiving antifungal therapy were analysed.; ResultsA total of 391 SNPs were detected, among which 6 coding SNPs were reported for the first time. Novel genetic alterations were detected in both azole and echinocandin resistance genes. A C. glabrata strain, which was resistant to echinocandins but highly susceptible to azoles, harboured an FKS2 S663P mutation plus a novel presumed loss-of-function CgPDR1 mutation. This isolate was from a patient with deep-seated and urinary candidiasis. Another C. glabrata isolate, with an MDR phenotype, carried a new FKS2 S663A mutation and a new putative gain-of-function CgPDR1 mutation (T370I); this isolate showed mutated (80%) and WT (20%) populations and was collected after 75 days of exposure to caspofungin from a patient who underwent complicated abdominal surgery.; ConclusionsThis study shows that NGS can be used for extensive assessment of genetic mutations involved in antifungal resistance. This type of wide genome approach will become very valuable for detecting mechanisms of resistance in clinical strains subjected to multidrug pressure.

Published
2015
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13. Development of echinocandin resistance in Candida krusei isolates following exposure to micafungin and caspofungin in a BM transplant unit

Author

E. Tavernier, M Desnos-Ollivier, Karine Augeul-Meunier, D. Guyotat, Jérôme Cornillon, Micha Srour, Amandine Fayard, F Honeyman, H. Raberin, Institut de Cancérologie Lucien Neuwirth, CHU Saint-Etienne, Université Jean Monnet [Saint-Étienne] (UJM), Centre National de Référence des Mycoses invasives et antifongiques - Mycologie moléculaire (CNRMA), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Hôtes, vecteurs et agents infectieux : biologie et dynamique (HVAIBD), Centre Hospitalier Universitaire de Saint-Etienne (CHU de Saint-Etienne), We thank Catherine Blanc and Damien Hoinard for technical assistance. We thank the Platform Genotyping of Pathogen and Public Health, Institut Pasteur, for sequencing facility., Centre Hospitalier Universitaire de Saint-Etienne [CHU Saint-Etienne] (CHU ST-E), Université Jean Monnet - Saint-Étienne (UJM), and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Male, Antifungal Agents, Echinocandins, chemistry.chemical_compound, MESH: Lipopeptides, Caspofungin, polycyclic compounds, MESH: Bone Marrow Transplantation, ComputingMilieux_MISCELLANEOUS, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, Bone Marrow Transplantation, Candida, MESH: Aged, 0303 health sciences, MESH: Middle Aged, biology, Candidiasis, Hematology, Middle Aged, MESH: Candidiasis, 3. Good health, MESH: Caspofungin, Hematologic Neoplasms, ComputingMethodologies_DOCUMENTANDTEXTPROCESSING, Female, lipids (amino acids, peptides, and proteins), medicine.drug, Adult, medicine.medical_specialty, Adolescent, Echinocandin, Microbiology, MESH: Drug Resistance, Fungal, Lipopeptides, 03 medical and health sciences, Drug Resistance, Fungal, MESH: Candida, Candida krusei, medicine, Humans, Intensive care medicine, Aged, 030304 developmental biology, MESH: Adolescent, Transplantation, MESH: Humans, 030306 microbiology, business.industry, MESH: Echinocandins, Micafungin, MESH: Adult, biochemical phenomena, metabolism, and nutrition, bacterial infections and mycoses, biology.organism_classification, MESH: Antifungal Agents, MESH: Male, stomatognathic diseases, chemistry, business, MESH: Female, MESH: Micafungin, MESH: Hematologic Neoplasms
Abstract

Development of echinocandin resistance in Candida krusei isolates following exposure to micafungin and caspofungin in a BM transplant unit

Published
2015
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14. Systematic phenotyping of a large-scale Candida glabrata deletion collection reveals novel antifungal tolerance genes

Author

Biao Ma, Walter Glaser, Dominique Ferrandon, Marina Marcet-Houben, Ken Haynes, Jessica Quintin, Ute Zeidler, Fabian Istel, Brendan P. Cormack, Ingrid E. Frohner, Sascha Brunke, Katja Seider, Christophe d'Enfert, Brian Green, Sophie Bachellier-Bassi, Lauren Ames, Steffen Rupp, Karl Kuchler, Ilse D. Jacobsen, Ekkehard Hiller, Helmut Jungwirth, Vitor Cabral, Michael Tscherner, Arnaud Firon, Tobias Schwarzmüller, Murielle Chauvel, Bernhard Hube, Toni Gabaldón, Publica, Medizinische Universität Wien = Medical University of Vienna, Johns Hopkins University School of Medicine [Baltimore], Imperial College London, Fraunhofer Institute for Interfacial Engineering and Biotechnology (IGB), Center for sepsis control care, Friedrich Schiller University Jena [Jena, Germany], Leibniz Institute for Natural Product Research and Infection Biology (Hans Knoell Institute), Friedrich-Schiller-Universität Jena, University of Exeter, Biologie et Pathogénicité fongiques, Institut National de la Recherche Agronomique (INRA)-Institut Pasteur [Paris], Cellule Pasteur UPMC, Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut Pasteur [Paris], Centre for Genomic Regulation [Barcelona] (CRG), Universitat Pompeu Fabra [Barcelona]-Centro Nacional de Analisis Genomico [Barcelona] (CNAG), Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut für Molekulare Biowissenschaften [Graz], Karl-Franzens-Universität Graz, This work was supported by the Austrian Science Foundation FWF through the ERA-Net Pathogenomics project FunPath to KK (FWF-API-0125), SR (BMBF: 0313931A), TG, CD, DF and BH, and in part by the Christian Doppler Society to KK. BC and BM were supported in part by NIH RO1AI46223. BH and SB were supported by the German Federal Ministry of Education and Research (BMBF) through FKZ: 01EO1002. TG was funded in part by grants from the Spanish ministry of science and innovation (GEN2006-27784E, BFU2009-09168). KH was funded by the BBSRC (BB/F005210/1). DF was also partly supported by CNRS and the Fondation pour la recherche Medicale (Programme Equipe FRM). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript., ANR-06-PATH-0005,FUNPATH,Genomic approaches to unravel the molecular mechanisms of pathogenicity in the human fungal pathogen candida glabrata(2006), Fraunhofer Institute for Interfacial Engineering and Biotechnology (Fraunhofer IGB), Fraunhofer (Fraunhofer-Gesellschaft), Friedrich-Schiller-Universität = Friedrich Schiller University Jena [Jena, Germany], Biologie et Pathogénicité fongiques (BPF), Institut National de la Recherche Agronomique (INRA)-Institut Pasteur [Paris] (IP), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut Pasteur [Paris] (IP), Universitat Pompeu Fabra [Barcelona] (UPF)-Centro Nacional de Analisis Genomico [Barcelona] (CNAG), Institut Pasteur [Paris]-Institut National de la Recherche Agronomique (INRA), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA), lenormand, christelle, and Programme de recherche transnational Pathogenomics (ERA-NET) - Genomic approaches to unravel the molecular mechanisms of pathogenicity in the human fungal pathogen candida glabrata - - FUNPATH2006 - ANR-06-PATH-0005 - PATHO - VALID

Subjects
Azoles, Antifungal Agents, Antifungal drug, Candida glabrata, Biochemistry, biofilm, Echinocandins, Gene Knockout Techniques, chemistry.chemical_compound, MESH: Lipopeptides, Caspofungin, Cell Wall, MESH: Azoles, saccharomyces cerevisiae, délétion, Biology (General), Candida albicans, MESH: Candida glabrata, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, MESH: Gene Knockout Techniques, 0303 health sciences, Fungal protein, MESH: Microbial Sensitivity Tests, biology, Candidiasis, Fungal genetics, MESH: Candidiasis, phénotype, Phenotype, Medical Microbiology, antifongique, MESH: Fungal Proteins, paroi cellulaire, Functional genomics, Research Article, medicine.drug, Echinocandin, QH301-705.5, Immunology, Microbial Sensitivity Tests, MESH: Biofilms, MESH: Phenotype, Microbiology, MESH: Drug Resistance, Fungal, Fungal Proteins, Lipopeptides, pathogène fongique, 03 medical and health sciences, MESH: Cell Wall, Drug Resistance, Fungal, Osmotic Pressure, Virology, MESH: Gene Library, Genetics, medicine, azole, Microbial Pathogens, Molecular Biology, Gene Library, 030304 developmental biology, 030306 microbiology, MESH: Echinocandins, Biology and Life Sciences, RC581-607, MESH: Osmotic Pressure, biology.organism_classification, bacterial infections and mycoses, MESH: Antifungal Agents, [SDV.MP.MYC] Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, chemistry, MESH: Gene Deletion, Biofilms, Parasitology, Immunologic diseases. Allergy, Gene Deletion
Abstract

The opportunistic fungal pathogen Candida glabrata is a frequent cause of candidiasis, causing infections ranging from superficial to life-threatening disseminated disease. The inherent tolerance of C. glabrata to azole drugs makes this pathogen a serious clinical threat. To identify novel genes implicated in antifungal drug tolerance, we have constructed a large-scale C. glabrata deletion library consisting of 619 unique, individually bar-coded mutant strains, each lacking one specific gene, all together representing almost 12% of the genome. Functional analysis of this library in a series of phenotypic and fitness assays identified numerous genes required for growth of C. glabrata under normal or specific stress conditions, as well as a number of novel genes involved in tolerance to clinically important antifungal drugs such as azoles and echinocandins. We identified 38 deletion strains displaying strongly increased susceptibility to caspofungin, 28 of which encoding proteins that have not previously been linked to echinocandin tolerance. Our results demonstrate the potential of the C. glabrata mutant collection as a valuable resource in functional genomics studies of this important fungal pathogen of humans, and to facilitate the identification of putative novel antifungal drug target and virulence genes., Author Summary Clinical infections by the yeast-like pathogen Candida glabrata have been ever-increasing over the past years. Importantly, C. glabrata is one of the most prevalent causes of drug-refractory fungal infections in humans. We have generated a novel large-scale collection encompassing 619 bar-coded C. glabrata mutants, each lacking a single gene. Extensive profiling of phenotypes reveals a number of novel genes implicated in tolerance to antifungal drugs that interfere with proper cell wall function, as well as genes affecting fitness of C. glabrata both during normal growth and under environmental stress. This fungal deletion collection will be a valuable resource for the community to study mechanisms of virulence and antifungal drug tolerance in C. glabrata, which is particularly relevant in view of the increasing prevalence of infections caused by this important human fungal pathogen.

Published
2014
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15. Candida spp. with acquired echinocandin resistance, France, 2004-2010

Author

Dannaoui , Eric, Desnos-Ollivier , Marie, Garcia-Hermoso , Dea, Grenouillet , Fredéric, Cassaing , Sophie, Baixench , Marie-Thérèse, Bretagne , Stéphane, Dromer , Françoise, Lortholary , Olivier, Renseigné , Non, Mycologie moléculaire, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Centre National de Référence Mycologie et Antifongiques-Mycologie Moléculaire (CNRMA), Institut Pasteur [Paris] (IP), Laboratoire Chrono-environnement (UMR 6249) (LCE), Centre National de la Recherche Scientifique (CNRS)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Centre Hospitalier Universitaire de Toulouse (CHU Toulouse), Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Institut Pasteur and Institut de Veille Sanitaire provided financial support for this study. E.D. has received funds for speaking from Merck and Schering, French Mycoses Study Group, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris], Laboratoire Chrono-environnement - CNRS - UBFC (UMR 6249) (LCE), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique ( CNRS ), Centre National de Référence Mycologie et Antifongiques-Mycologie Moléculaire ( CNRMA ), Laboratoire Chrono-environnement ( LCE ), Université Bourgogne Franche-Comté ( UBFC ) -Centre National de la Recherche Scientifique ( CNRS ) -Université de Franche-Comté ( UFC ), Centre Hospitalier Universitaire de Toulouse, CHU Toulouse [Toulouse], and CHU Cochin [AP-HP]

Subjects
Male, Antifungal Agents, MESH : Aged, Candida glabrata, MESH : Candidiasis, MESH: Aged, 80 and over, Candida albicans, Candida krusei, polycyclic compounds, MESH : Female, caspofungin, [ SDV.MP.MYC ] Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, Candida, Aged, 80 and over, MESH: Aged, MESH: Microbial Sensitivity Tests, MESH: Middle Aged, Candidiasis, Dispatch, MESH : Adult, Middle Aged, MESH: Candidiasis, MESH: Young Adult, FKS mutation, Female, France, Adult, Adolescent, clinical isolates, MESH : Male, MESH : Antifungal Agents, education, MESH : Young Adult, Microbial Sensitivity Tests, MESH: Drug Resistance, Fungal, echinocandins, Young Adult, Drug Resistance, Fungal, MESH: Candida, MESH : Adolescent, Humans, MESH : Middle Aged, MESH : France, MESH : Aged, 80 and over, MESH : Echinocandins, Aged, treatment failure, MESH: Adolescent, drug resistance, MESH: Humans, MESH: Echinocandins, MESH : Humans, MESH : Drug Resistance, Fungal, MESH: Adult, bacterial infections and mycoses, MESH: Antifungal Agents, MESH: Male, MESH: France, genotyping, MESH : Microbial Sensitivity Tests, fungi, MESH: Female, MESH : Candida
Abstract

This work was presented in part at the 20th European Congress of Clinical Microbiology and Infectious Diseases, Vienna, Austria, April 10-13, 2010 (abstract no. O346); International audience; We report 20 episodes of infection caused by acquired echinocandin-resistant Candida spp. harboring diverse and new Fksp mutations. For 12 patients, initial isolates (low MIC, wild-type Fksp sequence) and subsequent isolates (after caspofungin treatment, high MIC, mutated Fksp) were genetically related.

Published
2012
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16. Chitin Synthases with a Myosin Motor-Like Domain Control the Resistance of Aspergillus fumigatus to Echinocandins

Author

Vishukumar Aimanianda, Isabelle Mouyna, Yves F. Dufrêne, Laetitia Muszkieta, Anne Beauvais, Sven Krappmann, Jean-Paul Latgé, Rémi Beau, David Alsteens, Cesar Roncero, Cristina Jiménez-Ortigosa, Stéphane Pire, UCL - SST/IMCN/BSMA - Bio and soft matter, Universidad de Salamanca, Aspergillus, Institut Pasteur [Paris], Université Catholique de Louvain = Catholic University of Louvain (UCL), Friedrich-Alexander Universität Erlangen-Nürnberg (FAU), Work in the Aspergillus unit was partly supported by European grants Fungwall, Antifun, and ESF Fuminomics. Work at the Université Catholique de Louvain was supported by the National Foundation for Scientific Research (FNRS), the Université Catholique de Louvain (Fonds Spéciaux de Recherche), the Région Wallonne, the Federal Office for Scientific, Technical and Cultural Affairs (Interuniversity Poles of Attraction Programme), and the Research Department of the Communauté Française de Belgique (Concerted Research Action). Y.F.D. and D.A. are Senior Research Associates and Postdoctoral Research Fellows, respectively, of the FRS-FNRS. Research at the laboratory of C.R. was supported by CICYT grants BIO2007-60779 and BFU2010-18632., European Project: 511952,FUNGWALL(2005), and Institut Pasteur [Paris] (IP)

Subjects
Antifungal Agents, Microscopy, Atomic Force, Aspergillus fumigatus, Echinocandins, chemistry.chemical_compound, Cell Wall, Gene Expression Regulation, Fungal, Pharmacology (medical), DNA, Fungal, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, chemistry.chemical_classification, MESH: Microscopy, Atomic Force, 0303 health sciences, MESH: Microbial Sensitivity Tests, biology, Strain (chemistry), MESH: Real-Time Polymerase Chain Reaction, Fungal genetics, Chitin synthase, Spores, Fungal, MESH: Mycelium, Phenotype, Infectious Diseases, Biochemistry, Glucosyltransferases, MESH: Aspergillus fumigatus, MESH: Gene Expression Regulation, Fungal, MESH: Carbohydrates, MESH: Mutation, Carbohydrates, Microbial Sensitivity Tests, macromolecular substances, Myosins, Real-Time Polymerase Chain Reaction, Polysaccharide, MESH: Phenotype, MESH: Drug Resistance, Fungal, Cell wall, 03 medical and health sciences, MESH: Cell Wall, Chitin, Mechanisms of Resistance, Drug Resistance, Fungal, Polysaccharides, MESH: Spores, Fungal, 030304 developmental biology, Chitin Synthase, Pharmacology, MESH: Glucosyltransferases, Mycelium, 030306 microbiology, MESH: Echinocandins, fungi, MESH: Chitin Synthase, MESH: Myosins, biology.organism_classification, MESH: Antifungal Agents, MESH: DNA, Fungal, MESH: Polysaccharides, chemistry, Mutation, biology.protein
Abstract

Aspergillus fumigatus has two chitin synthases ( CSMA and CSMB ) with a myosin motor-like domain (MMD) arranged in a head-to-head configuration. To understand the function of these chitin synthases, single and double csm mutant strains were constructed and analyzed. Although there was a slight reduction in mycelial growth of the mutants, the total chitin synthase activity and the cell wall chitin content were similar in the mycelium of all of the mutants and the parental strain. In the conidia, chitin content in the Δ csmA strain cell wall was less than half the amount found in the parental strain. In contrast, the Δ csmB mutant strain and, unexpectedly, the Δ csmA /Δ csmB mutant strain did not show any modification of chitin content in their conidial cell walls. In contrast to the hydrophobic conidia of the parental strain, conidia of all of the csm mutants were hydrophilic due to the presence of an amorphous material covering the hydrophobic surface-rodlet layer. The deletion of CSM genes also resulted in an increased susceptibility of resting and germinating conidia to echinocandins. These results show that the deletion of the CSMA and CSMB genes induced a significant disorganization of the cell wall structure, even though they contribute only weakly to the overall cell wall chitin synthesis.

Published
2012
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17. Global transcriptome changes underlying colony growth in the opportunistic human pathogen Aspergillus fumigatus

Author

Antonis Rokas, Jean-Paul Latgé, John G. Gibbons, Rémi Beau, Kriston L. McGary, Anne Beauvais, Department of Biological Sciences [Nashville], Vanderbilt University [Nashville], Aspergillus, Institut Pasteur [Paris], This work was conducted in part using the resources of the Advanced Computing Center for Research and Education at Vanderbilt University. J.G.G. is funded by the Graduate Program in Biological Sciences at Vanderbilt University and the National Institute of Allergy and Infectious Diseases, National Institutes of Health (NIH, NIAID: F31AI091343-01). The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIAID or the NIH. Work in J.-P.L.’sAspergillus lab is partly funded by the ESF grant Fuminomics and the ALLFUN FP7 project. Research in A.R.’s lab is supported by the Searle Scholars Program and the National Science Foundation (DEB-0844968), European Project: 260338,EC:FP7:HEALTH,FP7-HEALTH-2010-single-stage,ALLFUN(2010), and Institut Pasteur [Paris] (IP)

Subjects
Candidate gene, [SDV]Life Sciences [q-bio], Gene Expression, Genome, Aspergillus fumigatus, Transcriptome, Cell Wall, Gene Expression Regulation, Fungal, Gene Regulatory Networks, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, MESH: Gene Regulatory Networks, Genetics, 0303 health sciences, Chromosome Mapping, General Medicine, Articles, MESH: Transcription Factors, Multigene Family, MESH: Protein Biosynthesis, MESH: Glycolysis, MESH: Fungal Proteins, MESH: Aspergillus fumigatus, Glycolysis, Metabolic Networks and Pathways, MESH: Gene Expression Regulation, Fungal, MESH: Gene Expression, Context (language use), MESH: Biofilms, Biology, Microbiology, MESH: Drug Resistance, Fungal, Fungal Proteins, 03 medical and health sciences, MESH: Cell Wall, Drug Resistance, Fungal, Secondary metabolism, Molecular Biology, Gene, 030304 developmental biology, 030306 microbiology, MESH: Transcriptome, Biofilm, biology.organism_classification, MESH: Metabolic Networks and Pathways, Biofilms, Protein Biosynthesis, MESH: Multigene Family, MESH: Chromosome Mapping, Ribosomes, MESH: Ribosomes, Transcription Factors
Abstract

Aspergillus fumigatus is the most common and deadly pulmonary fungal infection worldwide. In the lung, the fungus usually forms a dense colony of filaments embedded in a polymeric extracellular matrix. To identify candidate genes involved in this biofilm (BF) growth, we used RNA-Seq to compare the transcriptomes of BF and liquid plankton (PL) growth. Sequencing and mapping of tens of millions sequence reads against the A. fumigatus transcriptome identified 3,728 differentially regulated genes in the two conditions. Although many of these genes, including the ones coding for transcription factors, stress response, the ribosome, and the translation machinery, likely reflect the different growth demands in the two conditions, our experiment also identified hundreds of candidate genes for the observed differences in morphology and pathobiology between BF and PL. We found an overrepresentation of upregulated genes in transport, secondary metabolism, and cell wall and surface functions. Furthermore, upregulated genes showed significant spatial structure across the A. fumigatus genome; they were more likely to occur in subtelomeric regions and colocalized in 27 genomic neighborhoods, many of which overlapped with known or candidate secondary metabolism gene clusters. We also identified 1,164 genes that were downregulated. This gene set was not spatially structured across the genome and was overrepresented in genes participating in primary metabolic functions, including carbon and amino acid metabolism. These results add valuable insight into the genetics of biofilm formation in A. fumigatus and other filamentous fungi and identify many relevant, in the context of biofilm biology, candidate genes for downstream functional experiments.

Published
2012
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18. Antifungal susceptibility profiles of 1698 yeast reference strains revealing potential emerging human pathogens

Author

Marie Desnos-Ollivier, Françoise Dromer, Vincent Robert, Dorothée Raoux-Barbot, Marizeth Groenewald, Centre National de Référence des Mycoses invasives et antifongiques - Mycologie moléculaire (CNRMA), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Centraalbureau voor Schimmelcultures Fungal Biodiversity Centre (CBS-KNAW), Thanks to Damien Hoinard (NRCMA) and Wendy Epping (CBS) for technical help. Thanks to Dea Garcia-Hermoso and Stéphane Bretagne for proofreading. Thanks to Jack Edwards and Marc Orbach for their helpful information about use of antifungal in agricultures., and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Basidiomycetes, Azoles, Antifungal Agents, Opportunistic pathogens, Antifungal drug, lcsh:Medicine, Human pathogen, MESH: Drug Design, Antimicrobial resistance, Reference Values, Candida albicans, MESH: Phylogeny, lcsh:Science, Phylogeny, Fungal pathogens, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, MESH: Microbial Sensitivity Tests, 0303 health sciences, Multidisciplinary, Ascomycota, biology, Temperature, MESH: Reference Values, MESH: Temperature, 3. Good health, Infectious Diseases, Phenotype, Medicine, Echinocandins, Research Article, MESH: Fungi, Drugs and Devices, Mycology, Microbial Sensitivity Tests, MESH: Ascomycota, MESH: Phenotype, Microbiology, MESH: Drug Resistance, Fungal, 03 medical and health sciences, MESH: Mycoses, Antibiotic resistance, Species Specificity, Drug Resistance, Fungal, In vivo, Humans, MESH: Species Specificity, Biology, 030304 developmental biology, Antifungals, MESH: Humans, 030306 microbiology, Basidiomycota, lcsh:R, Fungi, MESH: Antifungal Agents, biology.organism_classification, Ascomycetes, Yeast, MESH: Basidiomycota, Mycoses, Drug Design, lcsh:Q
Abstract

International audience; New molecular identification techniques and the increased number of patients with various immune defects or underlying conditions lead to the emergence and/or the description of novel species of human and animal fungal opportunistic pathogens. Antifungal susceptibility provides important information for ecological, epidemiological and therapeutic issues. The aim of this study was to assess the potential risk of the various species based on their antifungal drug resistance, keeping in mind the methodological limitations. Antifungal susceptibility profiles to the five classes of antifungal drugs (polyens, azoles, echinocandins, allylamines and antimetabolites) were determined for 1698 yeast reference strains belonging to 992 species (634 Ascomycetes and 358 Basidiomycetes). Interestingly, geometric mean minimum inhibitory concentrations (MICs) of all antifungal drugs tested were significantly higher for Basidiomycetes compared to Ascomycetes (p

Published
2012
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19. Pneumocystosis: a network survey in the Paris area 2003-2008

Author

P. Roux, Marie-Elisabeth Bougnoux, M. Cornet, C. Sarfati, Françoise Botterel, Claire Bouges-Michel, Patrice Bourée, Hélène Yera, G. Galeazzi, D. Magne, C. Chochillon, Adela Angoulvant, Eric Dannaoui, Arnaud Fekkar, CHU Saint-Antoine [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), CHU Tenon [AP-HP], Hôpital Henri Mondor, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Hôpital Avicenne [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), CHU Necker - Enfants Malades [AP-HP], AP-HP Hôpital Bicêtre (Le Kremlin-Bicêtre), AP-HP - Hôpital Bichat - Claude Bernard [Paris], Thérapeutique Recombinante Expérimentale (TIMC-IMAG-TheREx), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS), Hôpital Européen Georges Pompidou [APHP] (HEGP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO), CHU Pitié-Salpêtrière [AP-HP], Hôpital Louis Mourier - AP-HP [Colombes], Hôpital Cochin [AP-HP], Hôpital St Louis, Assistance Publique-Hôpitaux de Paris (AP-HP), Information – Technologies – Analyse Environnementale – Procédés Agricoles (UMR ITAP), Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Centre national du machinisme agricole, du génie rural, des eaux et forêts (CEMAGREF), Hôpital Bichat-Claude Bernard, Hôpital Hôtel Dieu, Hôpital Européen Georges Pompidou, Hôpital St Antoine, Assistance publique - Hôpitaux de Paris (AP-HP)-Université Paris Descartes - Paris 5 ( UPD5 ), Parasitogie-Mycologie, CHU Saint-Antoine [APHP], CHU Tenon [APHP], Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 ( UPEC UP12 ), Hôpital avicenne, Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Avicenne-Université Paris 13 ( UP13 ), CHU Pitié-Salpêtrière [APHP], Hopital Louis Mourier - AP-HP [Colombes], CHU Cochin [AP-HP], Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Assistance publique - Hôpitaux de Paris (AP-HP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Descartes - Paris 5 (UPD5), Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Université Pierre et Marie Curie - Paris 6 (UPMC)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Thérapeutique Recombinante Expérimentale ( TIMC-IMAG-THEREX ), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications [Grenoble] ( TIMC-IMAG ), Université Joseph Fourier - Grenoble 1 ( UJF ) -Institut polytechnique de Grenoble - Grenoble Institute of Technology ( Grenoble INP ) -IMAG-Centre National de la Recherche Scientifique ( CNRS ) -Université Grenoble Alpes ( UGA ) -Université Joseph Fourier - Grenoble 1 ( UJF ) -Institut polytechnique de Grenoble - Grenoble Institute of Technology ( Grenoble INP ) -IMAG-Centre National de la Recherche Scientifique ( CNRS ) -Université Grenoble Alpes ( UGA ), Information – Technologies – Analyse Environnementale – Procédés Agricoles ( UMR ITAP ), Institut national d’études supérieures agronomiques de Montpellier ( Montpellier SupAgro ) -Institut national de recherche en sciences et technologies pour l'environnement et l'agriculture ( IRSTEA ), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Université Paris Descartes - Paris 5 (UPD5), Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Université Paris 13 (UP13)-Hôpital Avicenne, Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP)-IMAG-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP)-IMAG-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), and Université Pierre et Marie Curie - Paris 6 (UPMC)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)

Subjects
Male, Antifungal Agents, MESH: CD4 Lymphocyte Count, MESH : Genotype, DHPS, HIV Infections, MESH: Comorbidity, Drug resistance, Comorbidity, Pneumocystis carinii, MESH: Genotype, 0302 clinical medicine, Adrenal Cortex Hormones, Neoplasms, Epidemiology, Pneumocystosis, MESH: Immunocompromised Host, MESH : Female, MESH: Neoplasms, MESH : Sulfanilamides, 030212 general & internal medicine, MESH: Dihydropteroate Synthase, [ SDV.MP.MYC ] Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, MESH : Pneumocystis jirovecii, 0303 health sciences, MESH: Middle Aged, Mortality rate, Pneumonia, Pneumocystis, MESH: Paris, MESH : Pneumonia, Pneumocystis, MESH : Paris, Life Sciences, MESH : Immunocompromised Host, General Medicine, MESH: HIV Infections, MESH: Transplantation, Middle Aged, MESH: Hospitals, Hospitals, 3. Good health, MESH : Hospitals, MESH : Antineoplastic Agents, Infectious Diseases, MESH : Comorbidity, Female, MESH : Dihydropteroate Synthase, Microbiology (medical), medicine.medical_specialty, Paris, Genotype, MESH : Male, MESH : Antifungal Agents, Antineoplastic Agents, MESH: Drug Resistance, Fungal, MESH : Adrenal Cortex Hormones, MESH: Adrenal Cortex Hormones, 03 medical and health sciences, Immunocompromised Host, MESH: Sulfanilamides, MESH: Pneumocystis jirovecii, Drug Resistance, Fungal, Internal medicine, Sulfanilamides, MESH : CD4 Lymphocyte Count, MESH : HIV Infections, medicine, Humans, MESH : Middle Aged, Mycosis, Dihydropteroate Synthase, Transplantation, MESH: Humans, 030306 microbiology, business.industry, MESH : Humans, MESH : Drug Resistance, Fungal, medicine.disease, MESH: Antifungal Agents, MESH : Neoplasms, MESH: Male, Surgery, CD4 Lymphocyte Count, MESH : Transplantation, MESH: Antineoplastic Agents, business, MESH: Female, MESH: Pneumonia, Pneumocystis
Abstract

International audience; The aims of this network group were to collect epidemiological data of PcP cases in 14 hospitals in the Paris area and to determine the Di-Hydro Pteroate Synthase (DHPS) genotypes, genetic markers for possible sulfamide resistance. From January 1, 2003 to December 31, 2008, 993 (mean 166/year) PcP cases have been reported. Sixty-five percent of patients were HIV-positive. The median count of CD4 lymphocytes was 32/mm(3) (30 in HIV-positive patients, 152 in HIV-negative patients). In HIV-positive patients, PcP revealed the HIV infection in 39%. Among 304 PcP occurring in HIV known infected patients, no prophylaxis was prescribed for 64%; cotrimoxazole prophylaxis had been prescribed to 47 patients but only one of them had the right compliance. In HIV-negative patients (264), corticosteroids were prescribed in 59% and cytotoxic chemotherapies in 34%; 78% did not receive prophylaxis. One hundred sixty nine tumoral pathologies and 116 transplantations were notified. The mortality rate was 16% at day 14 (13% in HIV-positive patients, 26% in HIV-negative patients). Mutations in DHPS genes were detected in 18.5% of samples; 12.5% of patients were infected with several strains. The total annual number of cases has been stable for five years but the proportion of HIV-negative patients increased from 25% to 43%.

Published
2011
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20. Recent exposure to caspofungin or fluconazole influences the epidemiology of candidemia: a prospective multicenter study involving 2,441 patients

Author

Karine Sitbon, Stéphane Bretagne, Françoise Dromer, Marie Desnos-Ollivier, Olivier Lortholary, Arnaud Fontanet, Centre de Référence Déficits Immunitaires Héréditaires (CEREDIH), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Necker - Enfants Malades [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Centre d'infectiologie Necker-Pasteur [CHU Necker], Institut Pasteur [Paris]-CHU Necker - Enfants Malades [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Mycologie moléculaire, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Centre National de Référence Mycologie et Antifongiques-Mycologie Moléculaire (CNRMA), Institut Pasteur [Paris], Epidémiologie des Maladies Emergentes - Emerging Diseases Epidemiology, Pasteur-Cnam Risques infectieux et émergents (PACRI), Institut Pasteur [Paris]-Conservatoire National des Arts et Métiers [CNAM] (CNAM)-Institut Pasteur [Paris]-Conservatoire National des Arts et Métiers [CNAM] (CNAM), Thérapeutique Recombinante Expérimentale (TIMC-IMAG-TheREx), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525 (TIMC-IMAG), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Institut Pasteur [Paris] (IP)-CHU Necker - Enfants Malades [AP-HP], Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP), Institut Pasteur [Paris] (IP)-Conservatoire National des Arts et Métiers [CNAM] (CNAM), HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-Institut Pasteur [Paris] (IP)-Conservatoire National des Arts et Métiers [CNAM] (CNAM), HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS), Centre de Référence Déficits Immunitaires Héréditaires ( CEREDIH ), Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Necker - Enfants Malades [AP-HP], CHU Necker - Enfants Malades [AP-HP]-Assistance publique - Hôpitaux de Paris (AP-HP), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique ( CNRS ), Centre National de Référence Mycologie et Antifongiques-Mycologie Moléculaire ( CNRMA ), Epidémiologie des Maladies Emergentes, Conservatoire National des Arts et Métiers [CNAM] ( CNAM ) -Institut Pasteur [Paris]-Pasteur-Cnam risques infectieux et émergents ( PACRI ), Institut Pasteur [Paris]-Conservatoire National des Arts et Métiers [CNAM] ( CNAM ), Thérapeutique Recombinante Expérimentale ( TIMC-IMAG-THEREX ), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications [Grenoble] ( TIMC-IMAG ), and Université Joseph Fourier - Grenoble 1 ( UJF ) -Institut polytechnique de Grenoble - Grenoble Institute of Technology ( Grenoble INP ) -IMAG-Centre National de la Recherche Scientifique ( CNRS ) -Université Grenoble Alpes ( UGA ) -Université Joseph Fourier - Grenoble 1 ( UJF ) -Institut polytechnique de Grenoble - Grenoble Institute of Technology ( Grenoble INP ) -IMAG-Centre National de la Recherche Scientifique ( CNRS ) -Université Grenoble Alpes ( UGA )

Subjects
Antifungal Agents, MESH : Prevalence, MESH : Prospective Studies, Drug resistance, MESH : Child, Preschool, MESH: Candidemia, law.invention, chemistry.chemical_compound, Echinocandins, 0302 clinical medicine, MESH : Child, law, Caspofungin, MESH: Child, Epidemiology, Candida albicans, MESH : Population Surveillance, Prevalence, Pharmacology (medical), 030212 general & internal medicine, Prospective Studies, Prospective cohort study, Child, Fluconazole, [ SDV.MP.MYC ] Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, Candida, 0303 health sciences, MESH: Microbial Sensitivity Tests, Cross Infection, MESH: Paris, MESH : Paris, MESH : Adult, Intensive care unit, 3. Good health, Infectious Diseases, Child, Preschool, Population Surveillance, MESH : Fluconazole, MESH: Fluconazole, France, MESH : Cross Infection, medicine.drug, Adult, medicine.medical_specialty, Paris, MESH : Candida albicans, MESH : Antifungal Agents, Microbial Sensitivity Tests, Biology, Clinical Therapeutics, MESH: Drug Resistance, Fungal, MESH: Population Surveillance, 03 medical and health sciences, Lipopeptides, Species Specificity, MESH: Candida, Drug Resistance, Fungal, Internal medicine, medicine, MESH : Species Specificity, MESH : Candidemia, MESH: Species Specificity, Humans, MESH : France, MESH: Prevalence, MESH : Echinocandins, Pharmacology, MESH: Humans, 030306 microbiology, MESH: Candida albicans, MESH: Echinocandins, MESH : Humans, MESH: Child, Preschool, MESH : Drug Resistance, Fungal, MESH: Cross Infection, Candidemia, MESH: Adult, Odds ratio, biology.organism_classification, MESH: Antifungal Agents, MESH: Prospective Studies, Surgery, MESH: France, chemistry, MESH : Microbial Sensitivity Tests, MESH : Candida
Abstract

A prospective multicenter surveillance program on yeast bloodstream infections was implemented in the Paris, France, area without restrictions on ward of hospitalization (intensive care unit, hematology, and surgery) or age (adults and children). The present analysis concerns 2,618 isolates collected over 7 years from 2,441 patients. Centralized species identification and antifungal susceptibility testing using the EUCAST methodology were performed. Almost 10% (232/2,441) of the patients had recently (≤30 days) been treated with antifungal drugs. We analyzed the effect of recent exposure to fluconazole ( n = 159) or caspofungin ( n = 61) on the proportions of the five major Candida species. For both drugs, preexposure was associated with a decreased prevalence of Candida albicans in favor of less drug-susceptible species ( C. glabrata and C. krusei for the former and C. parapsilosis and, to a lesser extent, C. glabrata and C. krusei for the latter; P = 0.001). In the multivariate analysis, the risk of being infected with an isolate with decreased susceptibility to fluconazole was independently associated with an age of ≥15 years (odds ratio [OR] = 2.45; 95% confidence interval [CI] = 1.39 to 4.31; P = 0.002) and with recent exposure to fluconazole (OR = 2.17; 95% CI = 1.51 to 3.13; P < 0.001), while the risk of being infected with an isolate with decreased susceptibility to caspofungin was independently associated with an age P = 0.001) and with recent exposure to caspofungin (OR = 4.79; 95% CI = 2.47 to 9.28; P < 0.001). These findings could influence future recommendations for the management of candidemia.

Published
2010
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21. Acquisition of flucytosine, azole, and caspofungin resistance in Candida glabrata bloodstream isolates serially obtained from a hematopoietic stem cell transplant recipient

Author

Nicolas Papon, Patricia Ribaud, Thierry Noël, Claire Lacroix, Gérard Socié, Aurélie Girard, Florence Chapeland-Leclerc, Christophe Hennequin, Microbiologie cellulaire et moléculaire et pathogénicité (MCMP), Université Bordeaux Segalen - Bordeaux 2-Centre National de la Recherche Scientifique (CNRS), Service greffe de moelle osseuse, Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Université Paris Diderot - Paris 7 (UPD7)-Groupe Hospitalier Saint Louis - Lariboisière - Fernand Widal [Paris], Assistance publique - Hôpitaux de Paris (AP-HP) (APHP), Université Paris Diderot - Paris 7 (UPD7)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Groupe Hospitalier Saint Louis - Lariboisière - Fernand Widal [Paris], and Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)

Subjects
Azoles, Antifungal Agents, medicine.medical_treatment, MESH: Flucytosine, Flucytosine, Candida glabrata, Hematopoietic stem cell transplantation, Drug resistance, chemistry.chemical_compound, Echinocandins, Caspofungin, MESH: Child, MESH: Azoles, Pharmacology (medical), Child, MESH: Candida glabrata, Fungemia, 0303 health sciences, Fungal protein, MESH: Microbial Sensitivity Tests, Candidiasis, Hematopoietic Stem Cell Transplantation, MESH: Candidiasis, 3. Good health, Infectious Diseases, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Female, MESH: Fungal Proteins, medicine.drug, Microbial Sensitivity Tests, Biology, Microbiology, MESH: Drug Resistance, Fungal, Fungal Proteins, 03 medical and health sciences, Lipopeptides, Drug Resistance, Fungal, MESH: Fungemia, medicine, Humans, Point Mutation, MESH: Hematopoietic Stem Cell Transplantation, 030304 developmental biology, MESH: Point Mutation, Pharmacology, MESH: Humans, 030306 microbiology, MESH: Echinocandins, biochemical phenomena, metabolism, and nutrition, medicine.disease, biology.organism_classification, bacterial infections and mycoses, MESH: Antifungal Agents, Transplantation, chemistry, Susceptibility, MESH: Female
Abstract

We describe the acquisition of flucytosine, azole, and caspofungin resistance in sequential Candida glabrata bloodstream isolates collected from a bone marrow transplant patient with clinical failure. Point mutations in C. glabrata FUR1 ( CgFUR1 ) and CgFKS2 and overexpression of CgCDR1 and CgCDR2 were observed in resistant isolates.

Published
2010
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22. A nonsense mutation in the ERG6 gene leads to reduced susceptibility to polyenes in a clinical isolate of Candida glabrata

Author

Jean-Philippe Bouchara, Thierry Bergès, Gérald Larcher, Patrick Vandeputte, Dominique Chabasse, Guy Tronchin, Emilie Ernoult, Groupe d'Étude des Interactions Hôte-Pathogène (GEIHP), Université d'Angers (UA), Physiologie Moléculaire du Transport des Sucres chez les Végétaux (PhyMoTS), and Université de Poitiers-Centre National de la Recherche Scientifique (CNRS)

Subjects
Azoles, Antifungal Agents, Candida glabrata, MESH: Base Sequence, medicine.disease_cause, MESH: Ergosterol, chemistry.chemical_compound, Plasmid, Ergosterol, MESH: Polyenes, MESH: Azoles, Pharmacology (medical), MESH: Codon, Nonsense, DNA, Fungal, MESH: Candida glabrata, 0303 health sciences, Mutation, biology, Candidiasis, Fungal genetics, MESH: Candidiasis, 3. Good health, Complementation, Infectious Diseases, Biochemistry, Codon, Nonsense, Guanine, Genes, Fungal, Molecular Sequence Data, Nonsense mutation, Polyenes, MESH: Drug Resistance, Fungal, 03 medical and health sciences, Pseudohyphal growth, Drug Resistance, Fungal, Mechanisms of Resistance, medicine, Humans, [SDV.BV]Life Sciences [q-bio]/Vegetal Biology, 030304 developmental biology, Pharmacology, MESH: Guanine, MESH: Humans, MESH: Molecular Sequence Data, Base Sequence, 030306 microbiology, biology.organism_classification, MESH: Antifungal Agents, Molecular biology, MESH: DNA, Fungal, chemistry, MESH: Genes, Fungal
Abstract

Unlike the molecular mechanisms that lead to azole drug resistance, the molecular mechanisms that lead to polyene resistance are poorly documented, especially in pathogenic yeasts. We investigated the molecular mechanisms responsible for the reduced susceptibility to polyenes of a clinical isolate of Candida glabrata . Sterol content was analyzed by gas-phase chromatography, and we determined the sequences and levels of expression of several genes involved in ergosterol biosynthesis. We also investigated the effects of the mutation harbored by this isolate on the morphology and ultrastructure of the cell, cell viability, and vitality and susceptibility to cell wall-perturbing agents. The isolate had a lower ergosterol content in its membranes than the wild type, and the lower ergosterol content was found to be associated with a nonsense mutation in the ERG6 gene and induction of the ergosterol biosynthesis pathway. Modifications of the cell wall were also seen, accompanied by increased susceptibility to cell wall-perturbing agents. Finally, this mutation, which resulted in a marked fitness cost, was associated with a higher rate of cell mortality. Wild-type properties were restored by complementation of the isolate with a centromeric plasmid containing a wild-type copy of the ERG6 gene. In conclusion, we have identified the molecular event responsible for decreased susceptibility to polyenes in a clinical isolate of C. glabrata . The nonsense mutation detected in the ERG6 gene of this isolate led to a decrease in ergosterol content. This isolate may constitute a useful tool for analysis of the relevance of protein trafficking in the phenomena of azole resistance and pseudohyphal growth.

Published
2008
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23. Molecular mechanism of flucytosine resistance in Candida lusitaniae: contribution of the FCY2, FCY1, and FUR1 genes to 5-fluorouracil and fluconazole cross-resistance

Author

Papon, Nicolas, Noël, Thierry, Florent, Martine, Gibot-Leclerc, Stéphanie, Jean, Dorothée, Chastin, Christiane, Villard, Jean, Chapeland-Leclerc, Florence, Laboratoire des Sciences de l'Image, de l'Informatique et de la Télédétection (LSIIT), and Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique (CNRS)

Subjects
MESH: Sequence Analysis, DNA, MESH: Mutation, Antifungal Agents, Molecular Sequence Data, MESH: Flucytosine, Flucytosine, Microbial Sensitivity Tests, MESH: Drug Resistance, Fungal, Cytosine Deaminase, Fungal Proteins, MESH: Plasmids, MESH: Candida, Mechanisms of Resistance, Drug Resistance, Fungal, MESH: Cloning, Molecular, Pentosyltransferases, Cloning, Molecular, DNA, Fungal, Fluconazole, Drug Resistance, Fungal/genetics, Fluconazole/pharmacology, Alleles, MESH: Pentosyltransferases, Candida, MESH: Genetic Complementation Test, MESH: Microbial Sensitivity Tests, Plasmids/genetics, MESH: Molecular Sequence Data, MESH: Alleles, MESH: Cytosine Deaminase, Genetic Complementation Test, Sequence Analysis, DNA, Pentosyltransferases/genetics, MESH: Antifungal Agents, DNA, Fungal/chemistry/genetics, MESH: DNA, Fungal, Flucytosine/pharmacology, Fungal Proteins/genetics, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Mutation, Antifungal Agents/pharmacology, Candida/drug effects/enzymology/genetics, Fluorouracil/pharmacology, MESH: Fluconazole, MESH: Fungal Proteins, Fluorouracil, Cytosine Deaminase/genetics, MESH: Fluorouracil, Plasmids
Abstract

International audience; Inactivation of the FCY2 (cytosine permease), FCY1 (cytosine deaminase), and FUR1 (uracil phosphoribosyltransferase) genes in Candida lusitaniae produced two patterns of resistance to flucytosine. Mutant fur1 demonstrated resistance to 5-fluorouracil, whereas mutants fcy1 and fcy2 demonstrated fluconazole resistance in the presence of subinhibitory flucytosine concentrations.

Published
2007
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24. [Antifungals cellular targets and mechanisms of resistance]

Author

Accoceberry, Isabelle, Noël, Thierry, Microbiologie cellulaire et moléculaire et pathogénicité (MCMP), Université Bordeaux Segalen - Bordeaux 2-Centre National de la Recherche Scientifique (CNRS), Laboratoire des Sciences de l'Image, de l'Informatique et de la Télédétection (LSIIT), and Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique (CNRS)

Subjects
MESH: Fungi, Azoles, Antifungal Agents, MESH: Echinocandins, MESH: Flucytosine, Fungi, MESH: Antibiotics, Antifungal, Flucytosine, MESH: Antifungal Agents, Peptides, Cyclic, MESH: Drug Resistance, Fungal, Echinocandins, Lipopeptides, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Caspofungin, Drug Resistance, Fungal, Amphotericin B, MESH: Amphotericin B, MESH: Azoles, MESH: Peptides, Cyclic
Abstract

International audience; Antifungals of systemic use for the treatment of invasive fungal infections belong to four main chemical families which have globally three cellular targets in fungal cells: fluorinated pyrimidines act on deoxyribonucleic acid (DNA) replication and protein synthesis; polyenes and azoles are toxic for ergosterol and its biosynthetic pathway; lipopeptides inhibit the synthesis of cell wall beta glucans. The resistance mechanisms that are developed by some fungi begin to be well understood particularly in Candida yeasts. The underlying bases of these mechanisms are either mutations that modify the antifungal target, or that block access to the target, and, on the other hand, the overexpression of genes encoding the target, or some membrane proteins involved in the active efflux of antifungal drugs.

Published
2006

25. [Surveillance of antimicrobial resistance and antimicrobial use in a university-affiliated hospital: implementation of a computerized system]

Author

Müller , A., Patry , I., Talon , D., Cornette , C., Lopez-Lozano , J. M., Plésiat , P., Bertrand , X., Max-Planck-Institut für Metallforschung and Institut für Nichtmetallische Anorganische Materialien, Max-Planck-Institut-Universität Stuttgart [Stuttgart], Physiopathologie du système nerveux., Université Louis Pasteur - Strasbourg I-IFR37-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ), Service de bactériologie [Besançon], Laboratoire Chrono-environnement ( LCE ), Université Bourgogne Franche-Comté ( UBFC ) -Centre National de la Recherche Scientifique ( CNRS ) -Université de Franche-Comté ( UFC ), Agents pathogènes et inflammation - UFC (EA 4266) ( API ), Université de Franche-Comté ( UFC ), Laboratoire Chrono-environnement - CNRS - UBFC (UMR 6249) (LCE), Centre National de la Recherche Scientifique (CNRS)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon), Agents pathogènes et inflammation - UFC (EA 4266) (API), and Université de Franche-Comté (UFC)

Subjects
MESH : Escherichia coli, MESH: Drug Resistance, Microbial, MESH : Drug Resistance, Bacterial, Hospitals, University, Anti-Infective Agents, [ SDV.MP ] Life Sciences [q-bio]/Microbiology and Parasitology, MESH: Staphylococcus aureus, MESH: Microbial Sensitivity Tests, MESH: Drug Resistance, Viral, MESH: Escherichia coli, MESH : Staphylococcus aureus, Drug Resistance, Microbial, Bacterial Infections, MESH : Penicillin Resistance, MESH : Bacterial Infections, MESH: Virus Diseases, MESH : Drug Resistance, Viral, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Virus Diseases, MESH : Anti-Infective Agents, France, MESH : Software, Staphylococcus aureus, MESH: Bacterial Infections, Penicillin Resistance, MESH: Anti-Infective Agents, Microbial Sensitivity Tests, MESH: Drug Resistance, Fungal, MESH: Software, MESH: Mycoses, Drug Resistance, Fungal, MESH : Drug Resistance, Microbial, Drug Resistance, Bacterial, Drug Resistance, Viral, MESH: Drug Resistance, Bacterial, Escherichia coli, Humans, MESH : France, MESH : Hospitals, University, MESH: Hospitals, University, MESH : Mycoses, MESH: Humans, MESH: Methicillin Resistance, MESH : Humans, MESH : Drug Resistance, Fungal, MESH: Penicillin Resistance, MESH : Virus Diseases, MESH : Methicillin Resistance, MESH: France, Mycoses, Methicillin Resistance, MESH : Microbial Sensitivity Tests, Software
Abstract

International audience; After half a century of antibiotic use, the increasing problem of the emergence and spread of antimicrobial-resistant pathogens has created a problem of public health. The causes of this problem are multifactorial, but the excessive and inappropriate use of antimicrobials is the principal cause. The current guidelines for the control of antimicrobial resistance in hospitals recommend the implementation of a surveillance system of antimicrobial use and antimicrobial resistance data. AIM OF THE STUDY: The objective of our project was to develop a computerised tool to survey the antibiotic consumption data and the antimicrobial resistance. MATERIALS ET METHODS: We have collected antimicrobial resistance data from the software of the bacteriology laboratory, antibiotic use data from the pharmacy and demographical data from the hospital's admission department. These data were integrated in a database server and available with a web application. Antimicrobial resistance data of 15 major microorganisms were extracted and expressed as a frequency with elimination of repeats by using time criteria (7, 14 or 28 days). Antibiotic use data were converted into defined daily doses (DDD) and expressed per 1000 patient-days. RESULTS: Data are available for consultation in the form of tables or graphs per unit, type of units (medicine, surgery, pediatrics, intensive care units) or in the whole hospital. The system allows the confrontation on the same graph of antimicrobial resistance and antibiotic use data. CONCLUSION: Our surveillance system constitutes a needed prerequisite to the implementation of a global strategy of antibiotic use improvement in our hospital.

Published
2006
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26. Biofilms and their role in the resistance of pathogenic Candida to antifungal agents

Author

Christophe d'Enfert, Biologie et Pathogénicité fongiques, Institut Pasteur [Paris]-Institut National de la Recherche Agronomique (INRA), Biologie et Pathogénicité fongiques (BPF), and Institut National de la Recherche Agronomique (INRA)-Institut Pasteur [Paris] (IP)

Subjects
Antifungal Agents, EFFLUX, Hypha, [SDV]Life Sciences [q-bio], Clinical Biochemistry, MULTI-DRUG RESISTANCE, MESH: Biofilms, PATHOGENICITE, Biology, ECHINOCANDIN, STEROL, MESH: Drug Resistance, Fungal, Microbiology, Extracellular matrix, Cell wall, 03 medical and health sciences, Drug Resistance, Fungal, MESH: Candida, Amphotericin B, Drug Discovery, medicine, Animals, Humans, MESH: Animals, Candida, 030304 developmental biology, Pharmacology, 0303 health sciences, MESH: Humans, 030306 microbiology, Biofilm, AZOLE, biochemical phenomena, metabolism, and nutrition, MESH: Antifungal Agents, Yeast, CASPOFUNGIN, 3. Good health, AMPHOTERICIN B, Biofilms, Molecular Medicine, Efflux, BIOFILM, Echinocandins, medicine.drug
Abstract

International audience; Fungal pathogens of the genus Candida form biofilms on catheters and prosthetic devices. These three-dimensional structures composed of yeast and hyphal cells embedded in an extracellular matrix constitute an important pitfall in the management of disseminated Candida infections because of their intrinsic resistance to almost all antifungals in clinical use. Candida biofilms are especially resistant to azoles and amphotericin B but remain sensitive to the newly introduced echinocandins that target cell wall beta-glucan biosynthesis. Antifungal resistance of biofilms results most probably from the conjunction of several mechanisms that act in a time-dependent manner. While drug efflux is likely to contribute to resistance during the early phases of biofilm formation, changes in the sterol composition of membranes might explain the resistance of mature biofilms. The original physiology of mature Candida biofilms is mirrored by specific gene expression patterns that may pinpoint genes important for the acquisition of pleiotropic antifungal resistance.

Published
2006
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27. The Fission Yeast Crb2/Chk1 Pathway Coordinates the DNA Damage and Spindle Checkpoint in Response to Replication Stress Induced by Topoisomerase I Inhibitor

Author

Ada Collura, Giuseppe Baldacci, Joël Blaisonneau, Stefania Francesconi, and Centre National de la Recherche Scientifique (CNRS)

Subjects
MESH: Signal Transduction, Cell cycle checkpoint, [SDV]Life Sciences [q-bio], Cell Cycle Proteins, MESH: Cell Cycle, MESH: DNA Replication, Gene Expression Regulation, Fungal, Enzyme Inhibitors, DNA, Fungal, Checkpoint Kinase 2, 0303 health sciences, Kinetochore, Cell Cycle, 030302 biochemistry & molecular biology, Nuclear Proteins, MESH: Chromosomes, Fungal, Cell biology, Spindle checkpoint, MESH: Topoisomerase I Inhibitors, MESH: Schizosaccharomyces, DNA Topoisomerases, Type I, MESH: Enzyme Inhibitors, Mad2 Proteins, MESH: Genome, Fungal, MESH: Camptothecin, Chromosomes, Fungal, Genome, Fungal, biological phenomena, cell phenomena, and immunity, MESH: Mad2 Proteins, MESH: Gene Expression Regulation, Fungal, Signal Transduction, DNA Replication, MESH: Mutation, DNA repair, Chromosome Structure and Dynamics, Spindle Apparatus, Protein Serine-Threonine Kinases, Topoisomerase-I Inhibitor, Biology, MESH: Checkpoint Kinase 2, MESH: Checkpoint Kinase 1, MESH: Protein-Serine-Threonine Kinases, MESH: Drug Resistance, Fungal, 03 medical and health sciences, MESH: Cell Cycle Proteins, Drug Resistance, Fungal, Schizosaccharomyces, CHEK1, MESH: Spindle Apparatus, Molecular Biology, MESH: Protein Kinases, Alleles, 030304 developmental biology, MESH: DNA Damage, MESH: Alleles, MESH: Schizosaccharomyces pombe Proteins, Cell Biology, G2-M DNA damage checkpoint, MESH: DNA, Fungal, Checkpoint Kinase 1, Mutation, Camptothecin, Schizosaccharomyces pombe Proteins, Topoisomerase I Inhibitors, Protein Kinases, MESH: Nuclear Proteins, MESH: DNA Topoisomerases, Type I, DNA Damage
Abstract

International audience; Living organisms experience constant threats that challenge their genome stability. The DNA damage checkpoint pathway coordinates cell cycle progression with DNA repair when DNA is damaged, thus ensuring faithful transmission of the genome. The spindle assembly checkpoint inhibits chromosome segregation until all chromosomes are properly attached to the spindle, ensuring accurate partition of the genetic material. Both the DNA damage and spindle checkpoint pathways participate in genome integrity. However, no clear connection between these two pathways has been described. Here, we analyze mutants in the BRCT domains of fission yeast Crb2, which mediates Chk1 activation, and provide evidence for a novel function of the Chk1 pathway. When the Crb2 mutants experience damaged replication forks upon inhibition of the religation activity of topoisomerase I, the Chk1 DNA damage pathway induces sustained activation of the spindle checkpoint, which in turn delays metaphase-to-anaphase transition in a Mad2-dependent fashion. This new pathway enhances cell survival and genome stability when cells undergo replicative stress in the absence of a proficient G(2)/M DNA damage checkpoint.

Published
2005
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28. Comparative Roles of the Cell Wall and Cell Membrane in Limiting Uptake of Xenobiotic Molecules by Saccharomyces cerevisiae

Author

Lluis M. Mir, Omar Tounekti, Mustapha Aouida, Omrane Belhadj, Vectorologie et transfert de gènes (VTG / UMR8121), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Biochimie et Technobiologie, Faculté des Sciences Mathématiques, Physiques et Naturelles de Tunis (FST), and Université de Tunis El Manar (UTM)-Université de Tunis El Manar (UTM)

Subjects
Cell Membrane Permeability, Spheroplasts, Cell membrane, Cell Wall, Pharmacology (medical), MESH: Spheroplasts, MESH: Cell Membrane Permeability, Internalization, media_common, 0303 health sciences, Fungal protein, Antibiotics, Antineoplastic, biology, 030302 biochemistry & molecular biology, Spheroplast, respiratory system, MESH: Saccharomyces cerevisiae, MESH: Bleomycin, 3. Good health, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], Infectious Diseases, Membrane, medicine.anatomical_structure, Electroporation, Biochemistry, MESH: Fungal Proteins, MESH: Membrane Proteins, media_common.quotation_subject, Saccharomyces cerevisiae, MESH: Drug Resistance, Fungal, Cell wall, Fungal Proteins, 03 medical and health sciences, Bleomycin, MESH: Cell Wall, Drug Resistance, Fungal, medicine, MESH: Electroporation, MESH: Antibiotics, Antineoplastic, Mechanisms of Action: Physiological Effects, 030304 developmental biology, Pharmacology, Cell Membrane, Membrane Proteins, biology.organism_classification, respiratory tract diseases, carbohydrates (lipids), Membrane protein, Biophysics, MESH: Cell Membrane
Abstract

Using reversible electropermeabilization of cells and spheroplasts, we show that the cell wall and plasma membrane partly account for bleomycin resistance by acting as two independent barriers. We also report on the presence of a membrane protein that may be responsible for bleomycin internalization and toxicity in Saccharomyces cerevisiae .

Published
2003

29. Pneumocystis jirovecii genotype associated with increased death rate of HIV-infected patients with pneumonia

Author

Rabodonirina, Meja, Vaillant, Laetitia, Taffé, Patrick, Nahimana, Aimable, Gillibert, René-Pierre, Vanhems, Philippe, Hauser, Philippe M., Institut Pasteur de Madagascar, Réseau International des Instituts Pasteur (RIIP), Service d'Hygiène, Epidémiologie et Prévention [Hôpital Edouard Herriot - HCL], Hôpital Edouard Herriot [CHU - HCL], Hospices Civils de Lyon (HCL)-Hospices Civils de Lyon (HCL), Laboratoire de Biométrie et Biologie Evolutive - UMR 5558 (LBBE), Université Claude Bernard Lyon 1 (UCBL), and Université de Lyon-Université de Lyon-Institut National de Recherche en Informatique et en Automatique (Inria)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Male, Epidemiology, HIV Infections, DHPS, Pneumocystis carinii, 0302 clinical medicine, MESH: Respiration, Artificial, MESH: Dihydropteroate Synthase, Aged, 80 and over, 0303 health sciences, Fungal protein, MESH: Middle Aged, Coinfection, Pneumonia, Pneumocystis, fungus, MESH: HIV Infections, antimicrobial drugs, Pneumocystis jirovecii pneumonia, MESH: Infant, 3. Good health, Survival Rate, immunocompromised, Child, Preschool, MESH: Fungal Proteins, Microbiology (medical), Article, lcsh:Infectious and parasitic diseases, 03 medical and health sciences, MESH: Pneumocystis jirovecii, pentamidine, Drug Resistance, Fungal, Humans, sulfa resistance, Aged, MESH: Adolescent, Dihydropteroate Synthase, MESH: Humans, 030306 microbiology, lcsh:R, MESH: Child, Preschool, Infant, MESH: Adult, medicine.disease, Respiration, Artificial, respiratory tract diseases, Pneumonia, chemistry, SMX/TMP, Mutation, Immunology, Dihydropteroate synthase, Pneumocystis jirovecii dihydropteroate synthase, MESH: Female, Pentamidine, MESH: Pneumonia, Pneumocystis, sulfamethoxazole/trimethoprim, opportunistic infection, [SDV]Life Sciences [q-bio], lcsh:Medicine, Dapsone, MESH: Trimethoprim-Sulfamethoxazole Combination, chemistry.chemical_compound, MESH: Aged, 80 and over, antibiotic, CME, antifungal drugs, intravenous drug use, Sulfamethoxazole/Trimethoprim, MESH: Aged, biology, homosexuality, Middle Aged, atovaquone, Infectious Diseases, fungal, Female, France, dihydropteroate synthase mutations, medicine.drug, Adult, MESH: Mutation, Adolescent, MESH: Survival Rate, 030231 tropical medicine, virus, MESH: Drug Resistance, Fungal, Fungal Proteins, mental disorders, Trimethoprim, Sulfamethoxazole Drug Combination, parasitic diseases, medicine, Pneumocystis jirovecii, lcsh:RC109-216, dapsone, Research, HIV, biology.organism_classification, MESH: Male, MESH: Coinfection, MESH: France, fungi
Abstract

Comorbidities might predict presence of specific fungal genotypes., Pneumocystis jirovecii dihydropteroate synthase (DHPS) mutations have been associated with failure of sulfa prophylaxis; their effect on the outcome of patients with P. jirovecii pneumonia (PCP) remains controversial. P. jirovecii DHPS polymorphisms and genotypes were identified in 112 cases of PCP in 110 HIV-infected patients by using PCR single-strand conformation polymorphism. Of the 110 patients observed, 21 died; 18 of those deaths were attributed to PCP. Thirty-three percent of the PCP cases involved a P. jirovecii strain that had 1 or both DHPS mutations. The presence or absence of DHPS mutations had no effect on the PCP mortality rate within 1 month, whereas P.jirovecii type 7 and mechanical ventilation at PCP diagnosis were associated with an increased risk of death caused by PCP. Mechanical ventilation at PCP diagnosis was also associated with an increased risk of sulfa treatment failure at 5 days.

Published
2013
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30. In vitro susceptibility of Aspergillus spp. clinical isolates to albendazole

Author

N. Berthet, O. Faure, P. Ambroise-Thomas, Jean-François Brugère, A. Bakri, Renée Grillot, Université Joseph Fourier - Grenoble 1 (UJF), Centre Hospitalier Universitaire [Grenoble] (CHU), Institut National de la Santé et de la Recherche Médicale (INSERM), We gratefully acknowledge Dr F. Symoens (IHEM, Brussels, Belgium) and Dr M. A. Piens (Laboratory of ‘Parasitologie-Mycologie et Maladies Tropicales’, Edouard Herriot Hospital, Lyon, France) for the gift of the strains. We are indebted to Dr B. Lebeau (‘Service de Parasitologie-Mycologie Médicale’, Grenoble University Hospital, France) for providing the other clinical strains. We are also grateful to SmithKline Beecham for the gift of albendazole., and BERTHET, Nicolas

Subjects
Microbiology (medical), Antifungal Agents, [SDV]Life Sciences [q-bio], Aspergillus flavus, Microbial Sensitivity Tests, Albendazole, drug susceptibility, benzimidazole, Aspergillus fumigatus, Microbiology, MESH: Drug Resistance, Fungal, 03 medical and health sciences, Aspergillus nidulans, Drug Resistance, Fungal, medicine, MESH: Albendazole, Humans, Pharmacology (medical), Aspergillus terreus, skin and connective tissue diseases, [SDV.MP] Life Sciences [q-bio]/Microbiology and Parasitology, 030304 developmental biology, Pharmacology, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, 0303 health sciences, Aspergillus, MESH: Microbial Sensitivity Tests, MESH: Humans, biology, 030306 microbiology, Aspergillus niger, Fungi imperfecti, biology.organism_classification, MESH: Antifungal Agents, 3. Good health, [SDV] Life Sciences [q-bio], Infectious Diseases, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, MESH: Aspergillus, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, antifungal, medicine.drug
Abstract

International audience; The in vitro antifungal activity of albendazole, a benzimidazole widely used as an antihelmintic drug in humans, was investigated and assessed for its activity against Aspergillus spp. Forty-eight isolates, representing the most frequent species found in human pathology [Aspergillus fumigatus (n = 27), Aspergillus flavus (n = 10), Aspergillus terreus (n = 7), Aspergillus nidulans (n = 3) and Aspergillus niger (n = 1)], and one quality control strain (A. niger ATCC 9804 83435) were tested according to the NCCLS M38-P methodology for moulds. All the strains were susceptible to albendazole, with homogeneous MICs for each species; three strains were resistant to itraconazole.

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