Candida albicans biofilm–induced vesicles confer drug resistance through matrix biogenesis

Zarnowski, Robert; Sanchez, Hiram; Covelli, Antonio S.; Dominguez, Eddie; Jaromin, Anna; Bernhardt, Jörg; Mitchell, Kaitlin F.; Heiss, Christian; Azadi, Parastoo; Mitchell, Aaron; Andes, David R.; Soll, David

doi:10.1371/journal.pbio.2006872

Candida albicans biofilm–induced vesicles confer drug resistance through matrix biogenesis

Journal Article · Mon Oct 08 00:00:00 EDT 2018 · PLoS Biology (Online)

DOI:https://doi.org/10.1371/journal.pbio.2006872· OSTI ID:1612320

Zarnowski, Robert ^[1]; Sanchez, Hiram ^[2]; Covelli, Antonio S. ^[2]; Dominguez, Eddie ^[2]; Jaromin, Anna ^[3]; Bernhardt, Jörg ^[4]; Mitchell, Kaitlin F. ^[2]; Heiss, Christian ^[5]; Azadi, Parastoo ^[5]; Mitchell, Aaron ^[6]; ^[2]; Soll, David ^[7]

Univ. of Wisconsin, Madison, WI (United States); DOE/OSTI
Univ. of Wisconsin, Madison, WI (United States)
Univ. of Wroclaw (Poland)
Ernst Moritz Arndt Univ. Greifswalk (Germany)
Univ. of Georgia, Athens, GA (United States)
Carnegie Mellon Univ., Pittsburgh, PA (United States)
Univ. of Iowa, Iowa City, IA (United States)

Cells from all kingdoms of life produce extracellular vesicles (EVs). Their cargo is protected from the environment by the surrounding lipid bilayer. EVs from many organisms have been shown to function in cell–cell communication, relaying signals that impact metazoan development, microbial quorum sensing, and pathogenic host–microbe interactions. Here, we have investigated the production and functional activities of EVs in a surface-associated microbial community or biofilm of the fungal pathogen Candida albicans. Crowded communities like biofilms are a context in which EVs are likely to function. Biofilms are noteworthy because they are encased in an extracellular polymeric matrix and because biofilm cells exhibit extreme tolerance to antimicrobial compounds. We found that biofilm EVs are distinct from those produced by free-living planktonic cells and display strong parallels in composition to biofilm matrix material. The functions of biofilm EVs were delineated with a panel of mutants defective in orthologs of endosomal sorting complexes required for transport (ESCRT) subunits, which are required for normal EV production in diverse eukaryotes. Most ESCRT-defective mutations caused reduced biofilm EV production, reduced matrix polysaccharide levels, and greatly increased sensitivity to the antifungal drug fluconazole. Matrix accumulation and drug hypersensitivity of ESCRT mutants were reversed by addition of wild-type (WT) biofilm EVs. Vesicle complementation showed that biofilm EV function derives from specific cargo proteins. Our studies indicate that C. albicans biofilm EVs have a pivotal role in matrix production and biofilm drug resistance. Biofilm matrix synthesis is a community enterprise; prior studies of mixed cell biofilms have demonstrated extracellular complementation. Therefore, EVs function not only in cell–cell communication but also in the sharing of microbial community resources.

View Accepted Manuscript (DOE)

Research Organization:: Univ. of Georgia, Athens, GA (United States)

Sponsoring Organization:: USDOE Office of Science (SC); National Institute of Allergy and Infectious Diseases (NIAID)

Grant/Contract Number:: SC0015662

OSTI ID:: 1612320

Journal Information:: PLoS Biology (Online), Journal Name: PLoS Biology (Online) Journal Issue: 10 Vol. 16; ISSN 1545-7885

Publisher:: Public Library of ScienceCopyright Statement

Country of Publication:: United States

Language:: English

References (51)

Probability-based protein identification by searching sequence databases using mass spectrometry data Perkins, David N.; Pappin, Darryl J. C.; Creasy, David M. Electrophoresis, Vol. 20, Issue 18 https://doi.org/10.1002/(SICI)1522-2683(19991201)20:18<3551::AID-ELPS3551>3.0.CO;2-2	journal	December 1999
Introduction to Extracellular Vesicles: Biogenesis, RNA Cargo Selection, Content, Release, and Uptake Abels, Erik R.; Breakefield, Xandra O. Cellular and Molecular Neurobiology, Vol. 36, Issue 3 https://doi.org/10.1007/s10571-016-0366-z	journal	April 2016
Detection of neutral and aminosugars from glycoproteins and polysaccharides as their alditol acetates Henry, Robert J.; Blakeney, Anthony B.; Harris, Philip J. Journal of Chromatography A, Vol. 256 https://doi.org/10.1016/S0021-9673(01)88259-5	journal	January 1983
Exosomes and Ectosomes in Intercellular Communication Meldolesi, Jacopo Current Biology, Vol. 28, Issue 8 https://doi.org/10.1016/j.cub.2018.01.059	journal	April 2018
Biogenesis and function of ESCRT-dependent extracellular vesicles Juan, Thomas; Fürthauer, Maximilian Seminars in Cell & Developmental Biology, Vol. 74 https://doi.org/10.1016/j.semcdb.2017.08.022	journal	February 2018
Imaging flow cytometry for the characterization of extracellular vesicles Lannigan, Joanne; Erdbruegger, Uta Methods, Vol. 112 https://doi.org/10.1016/j.ymeth.2016.09.018	journal	January 2017
Biofilms: Survival Mechanisms of Clinically Relevant Microorganisms Pugliese, Gina; Favero, Martin S. Infection Control & Hospital Epidemiology, Vol. 23, Issue 6 https://doi.org/10.1017/s0195941700082850	journal	June 2002
Subfemtomole MS and MS/MS Peptide Sequence Analysis Using Nano-HPLC Micro-ESI Fourier Transform Ion Cyclotron Resonance Mass Spectrometry Martin, Susan E.; Shabanowitz, Jeffrey; Hunt, Donald F. Analytical Chemistry, Vol. 72, Issue 18 https://doi.org/10.1021/ac000497v	journal	September 2000
The ESCRT machinery regulates the secretion and long-range activity of Hedgehog Matusek, Tamás; Wendler, Franz; Polès, Sophie Nature, Vol. 516, Issue 7529 https://doi.org/10.1038/nature13847	journal	December 2014
Large-scale production and isolation of Candida biofilm extracellular matrix Zarnowski, Robert; Sanchez, Hiram; Andes, David R. Nature Protocols, Vol. 11, Issue 12 https://doi.org/10.1038/nprot.2016.132	journal	October 2016
Targeting microbial biofilms: current and prospective therapeutic strategies Koo, Hyun; Allan, Raymond N.; Howlin, Robert P. Nature Reviews Microbiology, Vol. 15, Issue 12 https://doi.org/10.1038/nrmicro.2017.99	journal	September 2017
Through the wall: extracellular vesicles in Gram-positive bacteria, mycobacteria and fungi Brown, Lisa; Wolf, Julie M.; Prados-Rosales, Rafael Nature Reviews Microbiology, Vol. 13, Issue 10 https://doi.org/10.1038/nrmicro3480	journal	September 2015
Bacterial biofilms: from the Natural environment to infectious diseases Hall-Stoodley, Luanne; Costerton, J. William; Stoodley, Paul Nature Reviews Microbiology, Vol. 2, Issue 2, p. 95-108 https://doi.org/10.1038/nrmicro821	journal	February 2004
Extra virgin olive oil improved body weight and insulin sensitivity in high fat diet-induced obese LDLr−/−.Leiden mice without attenuation of steatohepatitis Álvarez-Amor, Leticia; Sierra, Amparo Luque; Cárdenas, Antonio Scientific Reports, Vol. 11, Issue 1 https://doi.org/10.1038/s41598-021-87761-3	journal	April 2021
Cutting-Edge Analysis of Extracellular Microparticles using ImageStreamX Imaging Flow Cytometry Headland, Sarah E.; Jones, Hefin R.; D'Sa, Adelina S. V. Scientific Reports, Vol. 4, Issue 1 https://doi.org/10.1038/srep05237	journal	June 2014
Chemical structure of the cell-wall mannan of Candida albicans serotype A and its difference in yeast and hyphal forms Shibata, Nobuyuki; Suzuki, Akifumi; Kobayashi, Hidemitsu Biochemical Journal, Vol. 404, Issue 3 https://doi.org/10.1042/BJ20070081	journal	May 2007
Community participation in biofilm matrix assembly and function Mitchell, Kaitlin F.; Zarnowski, Robert; Sanchez, Hiram Proceedings of the National Academy of Sciences, Vol. 112, Issue 13 https://doi.org/10.1073/pnas.1421437112	journal	March 2015
Liposomal Formulation of DIMIQ, Potential Antitumor Indolo[2,3- b ]Quinoline Agent and Its Cytotoxicity on Hepatoma Morris 5123 Cells Jaromin, Anna; Kozubek, Arkadiusz; Suchoszek-Lukaniuk, Katarzyna Drug Delivery, Vol. 15, Issue 1 https://doi.org/10.1080/10717540701829192	journal	January 2008
Candida biofilm: a well-designed protected environment Mukherjee, Pranab K.; Zhou, Guangyin; Munyon, Ryan Medical Mycology, Vol. 43, Issue 3 https://doi.org/10.1080/13693780500107554	journal	January 2005
Clinical Practice Guidelines for the Management of Candidiasis: 2009 Update by the Infectious Diseases Society of America Pappas, Peter G.; Kauffman, Carol A.; Andes, David Clinical Infectious Diseases, Vol. 48, Issue 5 https://doi.org/10.1086/596757	journal	March 2009
Genetic Basis of Candida Biofilm Resistance Due to Drug‐Sequestering Matrix Glucan Nett, Jeniel E.; Sanchez, Hiram; Cain, Michael T. The Journal of Infectious Diseases, Vol. 202, Issue 1 https://doi.org/10.1086/651200	journal	July 2010
Experimental Protein Mixture for Validating Tandem Mass Spectral Analysis Keller, Andrew; Purvine, Samuel; Nesvizhskii, Alexey I. OMICS: A Journal of Integrative Biology, Vol. 6, Issue 2 https://doi.org/10.1089/153623102760092805	journal	April 2002
Extracellular Vesicles: Composition, Biological Relevance, and Methods of Study Zaborowski, MikoŁaj P.; Balaj, Leonora; Breakefield, Xandra O. BioScience, Vol. 65, Issue 8 https://doi.org/10.1093/biosci/biv084	journal	June 2015
KEGG: Kyoto Encyclopedia of Genes and Genomes Kanehisa, Minoru; Goto, Susumu Nucleic Acids Research, Vol. 28, Issue 1, p. 27-30 https://doi.org/10.1093/nar/28.1.27	journal	January 2000
KEGG as a reference resource for gene and protein annotation Kanehisa, Minoru; Sato, Yoko; Kawashima, Masayuki Nucleic Acids Research, Vol. 44, Issue D1 https://doi.org/10.1093/nar/gkv1070	journal	October 2015
Compositional and immunobiological analyses of extracellular vesicles released by C andida albicans : Extracellular vesicles from Vargas, Gabriele; Rocha, Juliana D. B.; Oliveira, Debora Leite Cellular Microbiology, Vol. 17, Issue 3 https://doi.org/10.1111/cmi.12374	journal	November 2014
Vesicular transport in Histoplasma capsulatum : an effective mechanism for trans-cell wall transfer of proteins and lipids in ascomycetes Albuquerque, Priscila Costa; Nakayasu, Ernesto S.; Rodrigues, Marcio L. Cellular Microbiology, Vol. 10, Issue 8 https://doi.org/10.1111/j.1462-5822.2008.01160.x	journal	August 2008
Standardized Method for In Vitro Antifungal Susceptibility Testing of Candida albicansBiofilms Ramage, Gordon; Vande Walle, Kacy; Wickes, Brian L. Antimicrobial Agents and Chemotherapy, Vol. 45, Issue 9 https://doi.org/10.1128/AAC.45.9.2475-2479.2001	journal	September 2001
Extracellular Vesicles of the Hyperthermophilic Archaeon “Thermococcus onnurineus” NA1 ^T Choi, Dong Hee; Kwon, Yong Min; Chiura, Hiroshi Xavier Applied and Environmental Microbiology, Vol. 81, Issue 14 https://doi.org/10.1128/AEM.00428-15	journal	May 2015
Epidemiology of Invasive Candidiasis: a Persistent Public Health Problem Pfaller, M. A.; Diekema, D. J. Clinical Microbiology Reviews, Vol. 20, Issue 1 https://doi.org/10.1128/CMR.00029-06	journal	January 2007
Biofilms: Survival Mechanisms of Clinically Relevant Microorganisms Donlan, R. M.; Costerton, J. W. Clinical Microbiology Reviews, Vol. 15, Issue 2 https://doi.org/10.1128/CMR.15.2.167-193.2002	journal	April 2002
Candida Infections of Medical Devices Kojic, Erna M.; Darouiche, Rabih O. Clinical Microbiology Reviews, Vol. 17, Issue 2 https://doi.org/10.1128/CMR.17.2.255-267.2004	journal	April 2004
Requirement for Candida albicans Sun41 in Biofilm Formation and Virulence Norice, Carmelle T.; Smith, Frank J.; Solis, Norma Eukaryotic Cell, Vol. 6, Issue 11 https://doi.org/10.1128/EC.00314-07	journal	September 2007
Interface of Candida albicans Biofilm Matrix-Associated Drug Resistance and Cell Wall Integrity Regulation Nett, Jeniel E.; Sanchez, Hiram; Cain, Michael T. Eukaryotic Cell, Vol. 10, Issue 12 https://doi.org/10.1128/EC.05126-11	journal	June 2011
Strains and Strategies for Large-Scale Gene Deletion Studies of the Diploid Human Fungal Pathogen Candida albicans Noble, Suzanne M.; Johnson, Alexander D. Eukaryotic Cell, Vol. 4, Issue 2 https://doi.org/10.1128/EC.4.2.298-309.2005	journal	February 2005
Development and Characterization of an In Vivo Central Venous Catheter Candida albicans Biofilm Model Andes, D.; Nett, J.; Oschel, P. Infection and Immunity, Vol. 72, Issue 10 https://doi.org/10.1128/IAI.72.10.6023-6031.2004	journal	October 2004
To Build a Biofilm O'Toole, George A. Journal of Bacteriology, Vol. 185, Issue 9 https://doi.org/10.1128/JB.185.9.2687-2689.2003	journal	May 2003
Conservation and Divergence in the Candida Species Biofilm Matrix Mannan-Glucan Complex Structure, Function, and Genetic Control Dominguez, Eddie; Zarnowski, Robert; Sanchez, Hiram mBio, Vol. 9, Issue 2 https://doi.org/10.1128/mBio.00451-18	journal	April 2018
Novel Entries in a Fungal Biofilm Matrix Encyclopedia Zarnowski, R.; Westler, W. M.; Lacmbouh, G. A. mBio, Vol. 5, Issue 4 https://doi.org/10.1128/mBio.01333-14	journal	July 2014
Antibody Binding Alters the Characteristics and Contents of Extracellular Vesicles Released by Histoplasma capsulatum Matos Baltazar, Ludmila; Nakayasu, Ernesto S.; Sobreira, Tiago J. P. mSphere, Vol. 1, Issue 2 https://doi.org/10.1128/mSphere.00085-15	journal	March 2016
Implications of Type 2 Diabetes on Adolescent Reproductive Health Risk Downs, Julie S.; Arslanian, Silva; Bruine de Bruin, Wändi The Diabetes Educator, Vol. 36, Issue 6 https://doi.org/10.1177/0145721710383586	journal	October 2010
Novel entries in a fungal biofilm matrix encyclopedia. Zarnowski, Robert; Westler, William M.; Lacmbouh, Ghislain Ade Carnegie Mellon University https://doi.org/10.1184/r1/6103916	text	January 2014
Novel entries in a fungal biofilm matrix encyclopedia. Zarnowski, Robert; Westler, William M.; Lacmbouh, Ghislain Ade Carnegie Mellon University https://doi.org/10.1184/r1/6103916.v1	text	January 2014
Characterization of Yeast Extracellular Vesicles: Evidence for the Participation of Different Pathways of Cellular Traffic in Vesicle Biogenesis Oliveira, Débora L.; Nakayasu, Ernesto S.; Joffe, Luna S. PLoS ONE, Vol. 5, Issue 6 https://doi.org/10.1371/journal.pone.0011113	journal	June 2010
A Candida Biofilm-Induced Pathway for Matrix Glucan Delivery: Implications for Drug Resistance Taff, Heather T.; Nett, Jeniel E.; Zarnowski, Robert PLoS Pathogens, Vol. 8, Issue 8 https://doi.org/10.1371/journal.ppat.1002848	journal	August 2012
Fungal Super Glue: The Biofilm Matrix and Its Composition, Assembly, and Functions Mitchell, Kaitlin F.; Zarnowski, Robert; Andes, David R. PLOS Pathogens, Vol. 12, Issue 9 https://doi.org/10.1371/journal.ppat.1005828	journal	September 2016
Exosome secretion affects social motility in Trypanosoma brucei Eliaz, Dror; Kannan, Sriram; Shaked, Hadassa PLOS Pathogens, Vol. 13, Issue 3 https://doi.org/10.1371/journal.ppat.1006245	journal	March 2017
Adjusting for multiple testing when reporting research results: the Bonferroni vs Holm methods. Aickin, M.; Gensler, H. American Journal of Public Health, Vol. 86, Issue 5 https://doi.org/10.2105/AJPH.86.5.726	journal	May 1996
Mechanisms of Candida biofilm drug resistance Taff, Heather T.; Mitchell, Kaitlin F.; Edward, Jessica A. Future Microbiology, Vol. 8, Issue 10 https://doi.org/10.2217/fmb.13.101	journal	October 2013
Comparative analysis of Candida biofilm quantitation assays Taff, Heather T.; Nett, Jeniel E.; Andes, David R. Medical Mycology, Vol. 50, Issue 2 https://doi.org/10.3109/13693786.2011.580016	journal	February 2012
Minimal experimental requirements for definition of extracellular vesicles and their functions: a position statement from the International Society for Extracellular Vesicles Lötvall, Jan; Hill, Andrew F.; Hochberg, Fred Journal of Extracellular Vesicles, Vol. 3, Issue 1 https://doi.org/10.3402/jev.v3.26913	journal	January 2014

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Extracellular Vesicle-Mediated RNA Release in Histoplasma capsulatum Alves, Lysangela R.; Peres da Silva, Roberta; Sanchez, David A. mSphere, Vol. 4, Issue 2 https://doi.org/10.1128/msphere.00176-19	journal	March 2019
Recent advances in understanding Candida albicans hyphal growth Arkowitz, Robert A.; Bassilana, Martine F1000Research, Vol. 8 https://doi.org/10.12688/f1000research.18546.1	journal	May 2019
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