Coregulation of extracellular vesicle production and fluconazole susceptibility in Cryptococcus neoformans
Juliana Rizzo,
Adèle Trottier,
Frédérique Moyrand,
Jean-Yves Coppée,
Corinne Maufrais,
Ana Claudia G. Zimbres,
Thi Tuong Vi Dang,
Alexandre Alanio,
Marie Desnos-Ollivier,
Isabelle Mouyna,
Gérard Péhau-Arnaude,
Pierre-Henri Commere,
Sophie Novault,
Iuliana V. Ene,
Leonardo Nimrichter,
Marcio L. Rodrigues,
Guilhem Janbon
Affiliations
Juliana Rizzo
Institut Pasteur, Université Paris Cité, Unité Biologie des ARN des Pathogènes Fongiques , Paris, France
Adèle Trottier
Institut Pasteur, Université Paris Cité, Unité Biologie des ARN des Pathogènes Fongiques , Paris, France
Frédérique Moyrand
Institut Pasteur, Université Paris Cité, Unité Biologie des ARN des Pathogènes Fongiques , Paris, France
Jean-Yves Coppée
Institut Pasteur, Université Paris Cité, Unité Biologie des ARN des Pathogènes Fongiques , Paris, France
Corinne Maufrais
Institut Pasteur, Université Paris Cité, Unité Biologie des ARN des Pathogènes Fongiques , Paris, France
Ana Claudia G. Zimbres
Instituto de Microbiologia Paulo de Góes (IMPG), Universidade Federal do Rio de Janeiro , Rio de Janeiro, Brazil
Thi Tuong Vi Dang
Institut Pasteur, Université Paris Cité, Unité Biologie des ARN des Pathogènes Fongiques , Paris, France
Alexandre Alanio
Institut Pasteur, Université Paris Cité, Centre National de Référence Mycoses Invasives et Antifongiques, Groupe de recherche Mycologie Translationnelle, Département de Mycologie , Paris, France
Marie Desnos-Ollivier
Institut Pasteur, Université Paris Cité, Centre National de Référence Mycoses Invasives et Antifongiques, Groupe de recherche Mycologie Translationnelle, Département de Mycologie , Paris, France
Isabelle Mouyna
Institut Pasteur, Université Paris Cité, Unité Biologie des ARN des Pathogènes Fongiques , Paris, France
Gérard Péhau-Arnaude
Institut Pasteur, Université Paris Cité, Plateforme de Bio-Imagerie Ultrastructurale , Paris, France
Pierre-Henri Commere
Institut Pasteur, Université Paris Cité, Cytometry and Biomarkers , Paris, France
Sophie Novault
Institut Pasteur, Université Paris Cité, Cytometry and Biomarkers , Paris, France
Iuliana V. Ene
Institut Pasteur, Université Paris Cité, Fungal Heterogeneity Group , Paris, France
Leonardo Nimrichter
Instituto de Microbiologia Paulo de Góes (IMPG), Universidade Federal do Rio de Janeiro , Rio de Janeiro, Brazil
Marcio L. Rodrigues
Instituto de Microbiologia Paulo de Góes (IMPG), Universidade Federal do Rio de Janeiro , Rio de Janeiro, Brazil
Guilhem Janbon
Institut Pasteur, Université Paris Cité, Unité Biologie des ARN des Pathogènes Fongiques , Paris, France
ABSTRACT Resistance to fluconazole (FLC), the most widely used antifungal drug, is typically achieved by altering the azole drug target and/or drug efflux pumps. Recent reports have suggested a link between vesicular trafficking and antifungal resistance. Here, we identified novel Cryptococcus neoformans regulators of extracellular vesicle (EV) biogenesis that impact FLC resistance. In particular, the transcription factor Hap2 does not affect the expression of the drug target or efflux pumps, yet it impacts the cellular sterol profile. Subinhibitory FLC concentrations also downregulate EV production. Moreover, in vitro spontaneous FLC-resistant colonies showed altered EV production, and the acquisition of FLC resistance was associated with decreased EV production in clinical isolates. Finally, the reversion of FLC resistance was associated with increased EV production. These data suggest a model in which fungal cells can regulate EV production in place of regulating the drug target gene expression as a first line of defense against antifungal assault in this fungal pathogen. IMPORTANCE Extracellular vesicles (EVs) are membrane-enveloped particles that are released by cells into the extracellular space. Fungal EVs can mediate community interactions and biofilm formation, but their functions remain poorly understood. Here, we report the identification of the first regulators of EV production in the major fungal pathogen Cryptococcus neoformans. Surprisingly, we uncover a novel role of EVs in modulating antifungal drug resistance. Disruption of EV production was associated with altered lipid composition and changes in fluconazole susceptibility. Spontaneous azole-resistant mutants were deficient in EV production, while loss of resistance restored initial EV production levels. These findings were recapitulated in C. neoformans clinical isolates, indicating that azole resistance and EV production are coregulated in diverse strains. Our study reveals a new mechanism of drug resistance in which cells adapt to azole stress by modulating EV production.
