title: A multi-host approach for the systematic analysis of virulence factors in Cryptococcus neoformans.
abstract: A multi-host approach was followed to screen a library of 1201 signature-tagged deletion strains of Cryptococcus neoformans mutants to identify previously unknown virulence factors. The primary screen was performed using a Caenorhabditis elegans-C. neoformans infection assay. The hits among these strains were reconfirmed as less virulent than the wild type in the insect Galleria mellonella-C. neoformans infection assay. After this 2-stage screen, and to prioritize hits, we performed serial evaluations of the selected strains, using the C. elegans model. All hit strains identified through these studies were validated in a murine model of systemic cryptococcosis. Twelve strains were identified through a stepwise screening assay. Among them, 4 (CSN1201, SRE1, RDI1, and YLR243W) were previously discovered, providing proof of principle for this approach, while the role of the remaining 8 genes (CKS101, CNC5600, YOL003C, CND1850, MLH3, HAP502, MSL5, and CNA2580) were not previously described in cryptococcal virulence. The multi-host approach is an efficient method of studying the pathogenesis of C. neoformans. We used diverse model hosts, C. elegans, G. mellonella, and mice, with physiological differences and identified 12 genes associated with mammalian infection. Our approach may be suitable for large pathogenesis screens.
authors.affiliation: Division of Infectious diseases, Warren Alpert Medical School of Brown University, Providence, Rhode Island.
authors.forename: Athanasios
authors.initials: A
authors.lastname: Desalermos
authors.affiliation: Division of Infectious diseases, Warren Alpert Medical School of Brown University, Providence, Rhode Island.
authors.forename: Xiaojiang
authors.initials: X
authors.lastname: Tan
authors.affiliation: Division of Infectious diseases, Warren Alpert Medical School of Brown University, Providence, Rhode Island.
authors.forename: Rajmohan
authors.initials: R
authors.lastname: Rajamuthiah
authors.affiliation: Division of Infectious diseases, Warren Alpert Medical School of Brown University, Providence, Rhode Island.
authors.forename: Marios
authors.initials: M
authors.lastname: Arvanitis
authors.affiliation: Division of Infectious diseases, Warren Alpert Medical School of Brown University, Providence, Rhode Island.
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Division of Infectious diseases, Warren Alpert Medical School of Brown University, Providence, Rhode Island.
authors.forename: Dedong
authors.initials: D
authors.lastname: Li
authors.affiliation: Division of Infectious diseases, Warren Alpert Medical School of Brown University, Providence, Rhode Island.
authors.forename: Themistoklis K
authors.initials: TK
authors.lastname: Kourkoumpetis
authors.affiliation: Division of Infectious diseases, Warren Alpert Medical School of Brown University, Providence, Rhode Island.
authors.forename: Beth Burgwyn
authors.initials: BB
authors.lastname: Fuchs
authors.affiliation: Division of Infectious diseases, Warren Alpert Medical School of Brown University, Providence, Rhode Island.
authors.forename: Eleftherios
authors.initials: E
authors.lastname: Mylonakis
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000382:microbiology*; D003453:Cryptococcosis / Q000382:microbiology / Q000473:pathology; D003455:Cryptococcus neoformans / Q000235:genetics / Q000472:pathogenicity*; D004195:Disease Models, Animal; D005260:Female; D017353:Gene Deletion; D005820:Genetic Testing; D051379:Mice; D009036:Moths / Q000382:microbiology*; D037521:Virulence Factors / Q000032:analysis* / Q000235:genetics
chemical_list: D037521:Virulence Factors
keywords: Caenorhabditis elegans; Cryptococcus neoformans; Galleria mellonella; model host; pathogenesis; virulence
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