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Transcriptomics analysis of amphotericin B and lactoferrin drug synergy in Cryptococcus neoformans H99

ABSTRACT: Invasive fungal infections (IFIs) are difficult to treat. Few effective antifungal drugs are available and many have problems with toxicity, efficacy and drug-resistance. To overcome these challenges, existing therapies may be enhanced using more than one agent acting in synergy. Previously, we have found amphotericin B (AMB) and the iron chelator, lactoferrin (LF), were synergistic against Cryptococcus neoformans and Saccharomyces cerevisiae. This study investigates the mechanism of AMB+LF synergy using RNA-seq in Cryptococcus neoformans H99.

ORGANISM(S): Cryptococcus neoformans var. grubii H99

PROVIDER: GSE130375 | GEO | 2019/09/08

REPOSITORIES: GEO

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Transcriptome and network analyses in Saccharomyces cerevisiae reveal that amphotericin B and lactoferrin synergy disrupt metal homeostasis and stress response

Project description:Invasive fungal infections (IFIs) are difficult to treat. Few effective antifungal drugs are available and many have problems with toxicity, efficacy and drug-resistance. To overcome these challenges, existing therapies may be enhanced using more than one agent acting in synergy. Previously, we have found amphotericin B (AMB) and the iron chelator, lactoferrin (LF), were synergistic against Cryptococcus neoformans and Saccharomyces cerevisiae. This study investigates the mechanism of AMB+LF synergy, using RNA-seq and network analyses. Genes involved in iron homeostasis showed increased expression upon treatment with AMB alone. Unexpectedly, AMB+LF treatment did not lead to increased expression of iron or zinc homeostasis genes however we observed decreased expression of oxidative stress response genes. Addition of iron or zinc to AMB+LF treated cells did not rescue the synergy, supporting the likelihood that the mechanism of synergy involves more than iron and zinc chelation. We clustered genes based on patterns of co-expression and found by network analysis that many genes involved in iron and zinc homeostasis, which have dysregulated expression upon AMB+LF treatment, are targets of transcription factors Aft1p and Zap1p. Hypothesizing that these might play a key role in the synergistic response, knock-out mutants of Aft1 and Zap1 were tested for increased sensitivity to AMB and oxidative stress. Both mutants showed hypersensitivity towards these treatments. Our results suggest the mechanism of AMB+LF synergy involves disruption to oxidative stress response, in addition to chelation of iron and zinc. Since Zap1 is conserved in C. neoformans and contains a putative drug binding domain, we suggest novel Zap1 binding molecules could be combined with existing antifungals to serve as synergistic antifungal treatments for this species.

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Transcriptomics analysis of amphotericin B and lactoferrin drug synergy in Cryptococcus neoformans H99

Project description:Transcriptomics analysis of amphotericin B and lactoferrin drug synergy in Cryptococcus neoformans H99

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Ribosome profiling of Cryptococcus neoformans H99

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Whole genome sequencing of amphotericin B adaptors in Cryptocooccus neoformans

Project description:We exposed Cryptococcus neoformans lab strain H99 to amphtericin B and obtained some adaptors. We did sequencing of these adaptors.

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Cryptococcus neoformans strain TJ1854 derived amphotericin B adaptors

Project description:In this study, H99 is a wild type Cryptococcus neoformans strain. TJ1854 is a chromosome 4 disomy strain derived from H99. Approximately 1 million cells of TJ1854 were spread on YPD plate supplemented with 6ug/ml amphtericin B. Randomlyh 27 adaptors (TJ2487-TJ2513) were chosen. These adaptors were sequenced.

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Project description:In this project, H99 is the wild type Cryptococcus neoformans strain. TJ1843 is a strain with segmental disomy of chromosome 11. TJ1843 is a derivative of H99. Approximately 1 million cells of TJ1843 were spread on YPD plate supplemented with 8 ug/ml amphotericin B. Randomly 27 adaptors (TJ3409-TJ3435) were chosen. These adaptors were sequenced.

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