Genome-wide expression profiling of the response to short-term exposure to fluconazole in Cryptococcus neoformans serotype A

Florio, Ada Rita; Ferrari, Sélène; Carolis, Elena De; Torelli, Riccardo; Fadda, Giovanni; Sanguinetti, Maurizio; Sanglard, Dominique

doi:10.1186/1471-2180-11-97

Cited by 40 publications

(47 citation statements)

References 53 publications

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“…3B). We compared published work that examined the transcriptome responses of S. cerevisiae (Kuo et al, 2010) and C. neoformans (Florio et al, 2011) to FLC. We found that, while there was significant overlap in orthologous genes impacted in the two species, (p = 1.6 × 10 −3 ), there were also considerable differences: 67% of the genes with an altered response in C. neoformans whose orthologs in S. cerevisiae did not exhibit significant change, (Table S7) (Kuo et al, 2010).…”

Section: Resultsmentioning

confidence: 99%

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Unraveling the Biology of a Fungal Meningitis Pathogen Using Chemical Genetics

Brown

Nelson

VanderSluis

et al. 2014

Cell

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Summary The fungal meningitis pathogen Cryptococcus neoformans is a central driver of mortality in HIV/AIDS. We report a genome-scale chemical genetic data map for this pathogen that quantifies the impact of 439 small molecule challenges on 1448 gene knockouts. We identified chemical phenotypes for 83% of mutants screened and at least one genetic response for each compound. C. neoformans chemical-genetic responses are largely distinct from orthologous published profiles of Saccharomyces cerevisiae, demonstrating the importance of pathogen-centered studies. We used the chemical-genetic matrix to predict novel pathogenicity genes, infer compound mode-of-action, and to develop an algorithm, O2M, that predicts antifungal synergies. These predictions were experimentally validated, thereby identifying virulence genes, a molecule that triggers G2/M arrest and inhibits the Cdc25 phosphatase, and many compounds that synergize with the antifungal drug fluconazole. Our work establishes a chemical-genetic foundation for approaching an infection responsible for greater than one-third of AIDS-related deaths.

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Section: Resultsmentioning

confidence: 99%

“…Comparison of transcriptional response to FLC in S. cerevisiae (Kuo et al, 2010) and C. neoformans (Florio et al, 2011). Related to Fig.…”

Section: Supplementary Materialsmentioning

confidence: 99%

Unraveling the Biology of a Fungal Meningitis Pathogen Using Chemical Genetics

Brown

Nelson

VanderSluis

et al. 2014

Cell

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“…Furthermore, incubation with sub-inhibitory concentrations of fluconazole caused C. albicans to upregulate oxidative stress response genes and made the organism more resistant to killing by phagocytes [36]. Exposure to sub-inhibitory concentrations of fluconazole had a similar affect on C. neoformans [37]. Oxidative stress has also been postulated to be involved in the synergy of compounds with the azoles.…”

Section: Discussionmentioning

confidence: 99%

Identification and Characterization of Antifungal Compounds Using a Saccharomyces cerevisiae Reporter Bioassay

et al. 2012

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New antifungal drugs are urgently needed due to the currently limited selection, the emergence of drug resistance, and the toxicity of several commonly used drugs. To identify drug leads, we screened small molecules using a Saccharomyces cerevisiae reporter bioassay in which S. cerevisiae heterologously expresses Hik1, a group III hybrid histidine kinase (HHK) from Magnaporthe grisea. Group III HHKs are integral in fungal cell physiology, and highly conserved throughout this kingdom; they are absent in mammals, making them an attractive drug target. Our screen identified compounds 13 and 33, which showed robust activity against numerous fungal genera including Candida spp., Cryptococcus spp. and molds such as Aspergillus fumigatus and Rhizopus oryzae. Drug-resistant Candida albicans from patients were also highly susceptible to compounds 13 and 33. While the compounds do not act directly on HHKs, microarray analysis showed that compound 13 induced transcripts associated with oxidative stress, and compound 33, transcripts linked with heavy metal stress. Both compounds were highly active against C. albicans biofilm, in vitro and in vivo, and exerted synergy with fluconazole, which was inactive alone. Thus, we identified potent, broad-spectrum antifungal drug leads from a small molecule screen using a high-throughput, S. cerevisiae reporter bioassay.

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“…Sage was used in Cryptococcus to examine the transcriptome changes in response to temperature and iron concentration manipulations (Hu et al, 2008; Steen et al, 2002). Recently, progress in transcriptional techniques has allowed researchers to analyze gene expression profiling changes to antifungal drugs, iron depletion, CO2 and oxidative stress by DNA microarray analysis in Cryptococcus (Florio et al, 2011; Kim et al, 2010; Upadhya et al, 2013). These microarray analytic techniques have resulted in the identification of new transcription factors and regulatory proteins in signaling pathways (Chun et al, 2011; O'Meara et al, 2010).…”

Section: Genome-wide Analysismentioning

confidence: 99%

New technology and resources for cryptococcal research

Zhang¹,

Park²,

Williamson³

2015

Fungal Genetics and Biology

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Rapid advances in molecular biology and genome sequencing have enabled the generation of new technology and resources for cryptococcal research. RNAi-mediated specific gene knock down has become routine and more efficient by utilizing modified shRNA plasmids and convergent promoter RNAi constructs. This system was recently applied in a high-throughput screen to identify genes involved in host-pathogen interactions. Gene deletion efficiencies have also been improved by increasing rates of homologous recombination through a number of approaches, including a combination of double-joint PCR with split-marker transformation, the use of dominant selectable markers and the introduction of Cre-Loxp systems into Cryptococcus. Moreover, visualization of cryptococcal proteins has become more facile using fusions with codon-optimized fluorescent tags, such as green or red fluorescent proteins or, mCherry. Using recent genome-wide analytical tools, new transcriptional factors and regulatory proteins have been identified in novel virulence-related signaling pathways by employing microarray analysis, RNA-sequencing and proteomic analysis.

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Genome-wide expression profiling of the response to short-term exposure to fluconazole in Cryptococcus neoformans serotype A

Cited by 40 publications

References 53 publications

Unraveling the Biology of a Fungal Meningitis Pathogen Using Chemical Genetics

Unraveling the Biology of a Fungal Meningitis Pathogen Using Chemical Genetics

Identification and Characterization of Antifungal Compounds Using a Saccharomyces cerevisiae Reporter Bioassay

New technology and resources for cryptococcal research

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