The Cryptococcus neoformans Titan cell is an inducible and regulated morphotype underlying pathogenesis

Ivy M Dambuza, Thomas Drake, Ambre Chapuis, Xin Zhou, Joao Correia, Leanne Taylor-Smith, Nathalie Legrave, Tim Rasmussen, Matthew C. Fisher, Tihana Bicanic, Thomas S Harrison, Marcel Jaspars, Robin C May, Gordon D Brown, Raif Yuecel, Donna M MacCallum, Elizabeth Ballou (Corresponding Author)

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Abstract

Fungal cells change shape in response to environmental stimuli, and these morphogenic transitions drive pathogenesis and niche adaptation. For example, dimorphic fungi switch between yeast and hyphae in response to changing temperature. The basidiomycete Cryptococcus neoformans undergoes an unusual morphogenetic transition in the host lung from haploid yeast to large, highly polyploid cells termed Titan cells. Titan cells influence fungal interaction with host cells, including through increased drug resistance, altered cell size, and altered Pathogen Associated Molecular Pattern exposure. Despite the important role these cells play in pathogenesis, understanding the environmental stimuli that drive the morphological transition, and the molecular mechanisms underlying their unique biology, has been hampered by the lack of a reproducible in vitro induction system. Here we demonstrate reproducible in vitro Titan cell induction in response to environmental stimuli consistent with the host lung. In vitro Titan cells exhibit all the properties of in vivo generated Titan cells, the current gold standard, including altered capsule, cell wall, size, high mother cell ploidy, and aneuploid progeny. We identify the bacterial peptidoglycan subunit Muramyl Dipeptide as a serum compound associated with shift in cell size and ploidy, and demonstrate the capacity of bronchial lavage fluid and bacterial co-culture to induce Titanisation. Additionally, we demonstrate the capacity of our assay to identify established (cAMP/PKA) and previously undescribed (USV101) regulators of Titanisation in vitro. Finally, we investigate the Titanisation capacity of clinical isolates and their impact on disease outcome. Together, these findings provide new insight into the environmental stimuli and molecular mechanisms underlying the yeast-to-Titan transition and establish an essential in vitro model for the future characterization of this important morphotype.
Original languageEnglish
Article numbere1006978
Pages (from-to)1-28
Number of pages28
JournalPLoS Pathogens
Volume14
Issue number5
DOIs
Publication statusPublished - 18 May 2018

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Saturn
Cryptococcus neoformans
Cell Size
Yeasts
Ploidies
Acetylmuramyl-Alanyl-Isoglutamine
Lung
Basidiomycota
Polyploidy
Hyphae
Peptidoglycan
Cell Shape
Haploidy
Bronchoalveolar Lavage Fluid
Aneuploidy
Coculture Techniques
Drug Resistance
Cell Wall
Capsules
Fungi

Keywords

  • Cryptococcus neoformans
  • Titan cell
  • Pathogenesis

Cite this

Dambuza, I. M., Drake, T., Chapuis, A., Zhou, X., Correia, J., Taylor-Smith, L., ... Ballou, E. (2018). The Cryptococcus neoformans Titan cell is an inducible and regulated morphotype underlying pathogenesis. PLoS Pathogens, 14(5), 1-28. [e1006978]. https://doi.org/10.1371/journal.ppat.1006978

The Cryptococcus neoformans Titan cell is an inducible and regulated morphotype underlying pathogenesis. / Dambuza, Ivy M; Drake, Thomas ; Chapuis, Ambre; Zhou, Xin; Correia, Joao ; Taylor-Smith, Leanne ; Legrave, Nathalie; Rasmussen, Tim; Fisher, Matthew C. ; Bicanic, Tihana; Harrison, Thomas S ; Jaspars, Marcel; May, Robin C; Brown, Gordon D; Yuecel, Raif; MacCallum, Donna M; Ballou, Elizabeth (Corresponding Author).

In: PLoS Pathogens, Vol. 14, No. 5, e1006978, 18.05.2018, p. 1-28.

Research output: Contribution to journalArticle

Dambuza, IM, Drake, T, Chapuis, A, Zhou, X, Correia, J, Taylor-Smith, L, Legrave, N, Rasmussen, T, Fisher, MC, Bicanic, T, Harrison, TS, Jaspars, M, May, RC, Brown, GD, Yuecel, R, MacCallum, DM & Ballou, E 2018, 'The Cryptococcus neoformans Titan cell is an inducible and regulated morphotype underlying pathogenesis', PLoS Pathogens, vol. 14, no. 5, e1006978, pp. 1-28. https://doi.org/10.1371/journal.ppat.1006978
Dambuza IM, Drake T, Chapuis A, Zhou X, Correia J, Taylor-Smith L et al. The Cryptococcus neoformans Titan cell is an inducible and regulated morphotype underlying pathogenesis. PLoS Pathogens. 2018 May 18;14(5):1-28. e1006978. https://doi.org/10.1371/journal.ppat.1006978
Dambuza, Ivy M ; Drake, Thomas ; Chapuis, Ambre ; Zhou, Xin ; Correia, Joao ; Taylor-Smith, Leanne ; Legrave, Nathalie ; Rasmussen, Tim ; Fisher, Matthew C. ; Bicanic, Tihana ; Harrison, Thomas S ; Jaspars, Marcel ; May, Robin C ; Brown, Gordon D ; Yuecel, Raif ; MacCallum, Donna M ; Ballou, Elizabeth. / The Cryptococcus neoformans Titan cell is an inducible and regulated morphotype underlying pathogenesis. In: PLoS Pathogens. 2018 ; Vol. 14, No. 5. pp. 1-28.
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T1 - The Cryptococcus neoformans Titan cell is an inducible and regulated morphotype underlying pathogenesis

AU - Dambuza, Ivy M

AU - Drake, Thomas

AU - Chapuis, Ambre

AU - Zhou, Xin

AU - Correia, Joao

AU - Taylor-Smith, Leanne

AU - Legrave, Nathalie

AU - Rasmussen, Tim

AU - Fisher, Matthew C.

AU - Bicanic, Tihana

AU - Harrison, Thomas S

AU - Jaspars, Marcel

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AU - Yuecel, Raif

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N1 - We thank Prof Joseph Heitman and Prof Kirsten Nielsen for helpful discussions. We thank Dr Lukasz Kozubowski for feedback on the manuscript. We thank Prof Arturo Casadevall for the anti-capsule antibody 18B7. We are grateful to Drs Attila Bebes and Linda Duncan in the Iain Fraser Cytometry Centre (Aberdeen University), Delyth Reed, and Debbie Wilkinson, Lucinda Wight and Kevin MacKenzie in the Microscopy and Histology Core Facility (Aberdeen University) for their expert help with the cytometry and microscopy experiments. We are also grateful for the assistance of staff at the University of Aberdeen Medical Research Facility. Funding: ERB: This work was supported by the UK Biotechnology and Biological Research Council (http://www.bbsrc.ac.uk; BB/M014525/1). IMD: Wellcome Trust Strategic Award in Medical Mycology and Fungal Immunology (wellcome.ac.uk; 097377). AC, DMM: UK National Centre for the Replacement, Refinement and Reduction of Animals in Research (https://www.nc3rs.org.uk; NC/N002482/1). GDB: Wellcome Trust (wellcome.ac.uk; 102705). IMD, TD, AC, GDB, RY, and DMM: MRC Centre for Medical Mycology at the University of Aberdeen (https://www.mrc.ac.uk; www.abdn.ac.uk; MR/N006364/1). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Y1 - 2018/5/18

N2 - Fungal cells change shape in response to environmental stimuli, and these morphogenic transitions drive pathogenesis and niche adaptation. For example, dimorphic fungi switch between yeast and hyphae in response to changing temperature. The basidiomycete Cryptococcus neoformans undergoes an unusual morphogenetic transition in the host lung from haploid yeast to large, highly polyploid cells termed Titan cells. Titan cells influence fungal interaction with host cells, including through increased drug resistance, altered cell size, and altered Pathogen Associated Molecular Pattern exposure. Despite the important role these cells play in pathogenesis, understanding the environmental stimuli that drive the morphological transition, and the molecular mechanisms underlying their unique biology, has been hampered by the lack of a reproducible in vitro induction system. Here we demonstrate reproducible in vitro Titan cell induction in response to environmental stimuli consistent with the host lung. In vitro Titan cells exhibit all the properties of in vivo generated Titan cells, the current gold standard, including altered capsule, cell wall, size, high mother cell ploidy, and aneuploid progeny. We identify the bacterial peptidoglycan subunit Muramyl Dipeptide as a serum compound associated with shift in cell size and ploidy, and demonstrate the capacity of bronchial lavage fluid and bacterial co-culture to induce Titanisation. Additionally, we demonstrate the capacity of our assay to identify established (cAMP/PKA) and previously undescribed (USV101) regulators of Titanisation in vitro. Finally, we investigate the Titanisation capacity of clinical isolates and their impact on disease outcome. Together, these findings provide new insight into the environmental stimuli and molecular mechanisms underlying the yeast-to-Titan transition and establish an essential in vitro model for the future characterization of this important morphotype.

AB - Fungal cells change shape in response to environmental stimuli, and these morphogenic transitions drive pathogenesis and niche adaptation. For example, dimorphic fungi switch between yeast and hyphae in response to changing temperature. The basidiomycete Cryptococcus neoformans undergoes an unusual morphogenetic transition in the host lung from haploid yeast to large, highly polyploid cells termed Titan cells. Titan cells influence fungal interaction with host cells, including through increased drug resistance, altered cell size, and altered Pathogen Associated Molecular Pattern exposure. Despite the important role these cells play in pathogenesis, understanding the environmental stimuli that drive the morphological transition, and the molecular mechanisms underlying their unique biology, has been hampered by the lack of a reproducible in vitro induction system. Here we demonstrate reproducible in vitro Titan cell induction in response to environmental stimuli consistent with the host lung. In vitro Titan cells exhibit all the properties of in vivo generated Titan cells, the current gold standard, including altered capsule, cell wall, size, high mother cell ploidy, and aneuploid progeny. We identify the bacterial peptidoglycan subunit Muramyl Dipeptide as a serum compound associated with shift in cell size and ploidy, and demonstrate the capacity of bronchial lavage fluid and bacterial co-culture to induce Titanisation. Additionally, we demonstrate the capacity of our assay to identify established (cAMP/PKA) and previously undescribed (USV101) regulators of Titanisation in vitro. Finally, we investigate the Titanisation capacity of clinical isolates and their impact on disease outcome. Together, these findings provide new insight into the environmental stimuli and molecular mechanisms underlying the yeast-to-Titan transition and establish an essential in vitro model for the future characterization of this important morphotype.

KW - Cryptococcus neoformans

KW - Titan cell

KW - Pathogenesis

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DO - 10.1371/journal.ppat.1006978

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