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153. Identification of Aspergillus fumigatus multidrug transporter genes and their potential involvement in antifungal resistance.

Author

Meneau, Isabelle, Coste, Alix T., and Sanglard, Dominique

Abstract

Aspergillus fumigatus can cause severe fatal invasive aspergillosis in immunocompromised patients but is also found in the environment. A. fumigatus infections can be treated with antifungals agents among which azole and echinocandins. Resistance to the class of azoles has been reported not only from patient samples but also from environmental samples. Azole resistance mechanisms involve for most isolates alterations at the site of the azole target (cyp51A); however, a substantial number of isolates can also exhibit non-cyp51A-mediated mechanisms. We aimed here to identify novel A. fumigatus genes involved in azole resistance. For this purpose, we designed a functional complementation system of A. fumigatus cDNAs expressed in a Saccharomyces cerevisiae isolate lacking the ATP Binding Cassette (ABC) transporter PDR5 and that was therefore more azole-susceptible than the parent wild type. Several genes were recovered including two distinct ABC transporters (atrF, atrI) and a Major Facilitator transporter (mdrA), from which atrI (Afu3g07300) and mdrA (Afu1g13800) were not yet described. atrI mediated resistance to itraconazole and voriconazole, while atrF only to voriconazole in S. cerevisiae. Gene inactivation of each transporter in A. fumigatus indicated that the transporters were involved in the basal level of azole susceptibility. The expression of the transporters was addressed in clinical and environmental isolates with several azole resistance profiles. Our results show that atrI and mdrA tended to be expressed at higher levels than atrF in normal growth conditions. atrF was upregulated in 2/4 of azole-resistant environmental isolates and was the only gene with a significant association between transporter expression and azole resistance. In conclusion, this work showed the potential of complementation to identify functional transporters. The identified transporters were suggested to participate in azole resistance of A. fumigatus; however, this hypothesis will need further approaches to be verified. [ABSTRACT FROM AUTHOR]

Published
2016
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154. Direct Molecular Diagnosis of Aspergillosis and CYP51A Profiling from Respiratory Samples of French Patients.

Author

Yanan Zhao, Garnaud, Cécile, Brenier-Pinchart, Marie-Pierre, Thiébaut-Bertrand, Anne, Saint-Raymond, Christel, Camara, Boubou, Hamidfar, Rebecca, Cognet, Odile, Maubon, Danièle, Cornet, Muriel, Perlin, David S., Rodrigues, Fernando, and Sanglard, Dominique

Subjects
MOLECULAR diagnosis, ASPERGILLOSIS
Abstract

Background: Microbiological diagnosis of aspergillosis and triazole resistance is limited by poor culture yield. To better estimate this shortcoming, we compared culture and molecular detection of A. fumigatus in respiratory samples from French patients at risk for aspergillosis. Methods: A total of 97 respiratory samples including bronchoalveolar lavages (BAL), bronchial aspirates (BA), tracheal aspirates, sputa, pleural fluids, and lung biopsy were collected from 33 patients having invasive aspergillosis (n = 12), chronic pulmonary aspergillosis (n = 3), allergic bronchopulmonary aspergillosis (n = 7), or colonization (n = 11) and 28 controls. Each specimen was evaluated by culture, pan-Aspergillus qPCR, and CYP51A PCR and sequencing. Results: One A. flavus and 19 A. fumigatus with one multiazole resistant strain (5.3%) were cultured from 20 samples. Culture positivity was 62.5, 75, 42.9, and 15.8% in ABPA, CPA, IA, and colonized patients, respectively. Aspergillus detection rate was significantly higher by pan-Aspergillus qPCR than by culture in IA (90.5 vs. 42.9%; P < 0.05) and colonization group (73.7 vs. 15.8%; P < 0.05). The CYP51A PCR found one TR34/L98H along with 5 novel cyp51A mutations (4 non-synonymous and 1 promoter mutations), yet no association can be established currently between these novel mutations and azole resistance. The analysis of 11 matched pairs of BA and BAL samples found that 9/11 BA carried greater fungal load than BAL and CYP51A detection was more sensitive in BA than in BAL. Conclusion: Direct molecular detection of Aspergillus spp. and azole resistance markers are useful adjunct tools for comprehensive aspergillosis diagnosis. The observed superior diagnostic value of BAs to BAL fluids warrants more in-depth study. [ABSTRACT FROM AUTHOR]

Published
2016
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155. Induced Bacterial Cross-Resistance toward Host Antimicrobial Peptides: A Worrying Phenomenon.

Author

Fleitas, Osmel, Franco, Octávio L., Bauer, Margaret E., and Sanglard, Dominique

Subjects
DRUG resistance in bacteria, ANTIBIOTICS, ANTIMICROBIAL peptides
Abstract

Bacterial resistance to conventional antibiotics has reached alarming levels, threatening to return to the pre-antibiotic era. Therefore, the search for new antimicrobial compounds that overcome the resistance phenomenon has become a priority. Antimicrobial peptides (AMPs) appear as one of the most promising antibiotic medicines. However, in recent years several AMP-resistance mechanisms have been described. Moreover, the AMP-resistance phenomenon has become more complex due to its association with cross-resistance toward AMP effectors of the host innate immune system. In this context, the use of AMPs as a therapeutic option could be potentially hazardous, since bacteria could develop resistance toward our innate immune system. Here, we review the findings of major studies that deal with the AMP cross-resistance phenomenon. [ABSTRACT FROM AUTHOR]

Published
2016
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158. Sensing of mammalian IL-17A regulates fungal adaptation and virulence

Author

Zelante, Teresa, primary, Iannitti, Rossana G., additional, De Luca, Antonella, additional, Arroyo, Javier, additional, Blanco, Noelia, additional, Servillo, Giuseppe, additional, Sanglard, Dominique, additional, Reichard, Utz, additional, Palmer, Glen E., additional, Latgè, Jean-Paul, additional, Puccetti, Paolo, additional, and Romani, Luigina, additional

Published
2012
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159. The bZIP Transcription Factor Rca1p Is a Central Regulator of a Novel CO2 Sensing Pathway in Yeast

Author

Cottier, Fabien, primary, Raymond, Martine, additional, Kurzai, Oliver, additional, Bolstad, Marianne, additional, Leewattanapasuk, Worraanong, additional, Jiménez-López, Claudia, additional, Lorenz, Michael C., additional, Sanglard, Dominique, additional, Váchová, Libuše, additional, Pavelka, Norman, additional, Palková, Zdena, additional, and Mühlschlegel, Fritz A., additional

Published
2012
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162. Interrogation of Related Clinical Pan-Azole-Resistant Aspergillus fumigatus Strains: G138C, Y431C, and G434C Single Nucleotide Polymorphisms in cyp51A , Upregulation of cyp51A , and Integration and Activation of Transposon Atf1 in the cyp51A Promoter

Author

Albarrag, Ahmed M., primary, Anderson, Michael J., additional, Howard, Susan J., additional, Robson, Geoff D., additional, Warn, Peter A., additional, Sanglard, Dominique, additional, and Denning, David W., additional

Published
2011
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171. Doxorubicin induces drug efflux pumps inCandida albicans

Author

Kofla, Grzegorz, primary, Turner, Vincent, additional, Schulz, Bettina, additional, Storch, Ulrike, additional, Froelich, Daniela, additional, Rognon, Bénédicte, additional, Coste, Alix T., additional, Sanglard, Dominique, additional, and Ruhnke, Markus, additional

Published
2011
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178. MALDI-TOF MS-based drug susceptibility testing of pathogens: The example ofCandida albicansand fluconazole

Author

Marinach, Carine, primary, Alanio, Alexandre, additional, Palous, Martine, additional, Kwasek, St��phanie, additional, Fekkar, Arnaud, additional, Brossas, Jean-Yves, additional, Brun, Sophie, additional, Snounou, Georges, additional, Hennequin, Christophe, additional, Sanglard, Dominique, additional, Datry, Annick, additional, Golmard, Jean-Louis, additional, and Mazier, Dominique, additional

Published
2009
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180. Reliability of the Vitek 2 Yeast Susceptibility Test for Detection of In Vitro Resistance to Fluconazole and Voriconazole in Clinical Isolates of Candida albicans and Candida glabrata

Author

Posteraro, Brunella, primary, Martucci, Rosa, additional, La Sorda, Marilena, additional, Fiori, Barbara, additional, Sanglard, Dominique, additional, De Carolis, Elena, additional, Florio, Ada Rita, additional, Fadda, Giovanni, additional, and Sanguinetti, Maurizio, additional

Published
2009
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184. Adaptation of a Gaussia princeps Luciferase reporter system in Candida albicans for in vivo detection in the Galleria mellonella infection model.

Author

Delarze, Eric, Ischer, Françoise, Sanglard, Dominique, and Coste, Alix T

Subjects
LUCIFERASES, CANDIDA albicans, GALLERIA, MYCOSES, BACTERIAL cell walls, LABORATORY rats, DRUG efficacy
Abstract

For the past 10 years, mini-host models and in particular the greater wax mothGalleria mellonellahave tended to become a surrogate for murine models of fungal infection mainly due to cost, ethical constraints and ease of use. Thus, methods to better assess the fungal pathogenesis inG. mellonellaneed to be developed. In this study, we implemented the detection ofCandida albicanscells expressing theGaussia princepsluciferase in its cell wall in infected larvae ofG. mellonella. We demonstrated that detection and quantification of luminescence in the pulp of infected larvae is a reliable method to perform drug efficacy andC. albicansvirulence assays as compared to fungal burden assay. Since the linearity of the bioluminescent signal, as compared to the CFU counts, has a correlation of R2= 0.62 and that this method is twice faster and less labor intensive than classical fungal burden assays, it could be applied to large scale studies. We next visualized and followedC. albicansinfection in livingG. mellonellalarvae using a non-toxic and water-soluble coelenterazine formulation and a CCD camera that is commonly used for chemoluminescence signal detection. This work allowed us to follow for the first timeC. albicanscourse of infection inG. mellonelladuring 4 days. [ABSTRACT FROM PUBLISHER]

Published
2015
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