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

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

PROVIDER: PRJNA539938 | ENA |

REPOSITORIES: ENA

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

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DNA-seq of Cryptococcus neoformans H99 derived amphotericin B adaptors

Project description:Approximately 1 million cells of Cryptococcus neoformans lab strain H99 were spread on YPD plate supplmemented with 3ug/ml amphotericin B. Randomly 30 adaptors (TJ1832 - TJ1861) were chosen. The parent and all the 30 adaptors were sequenced.

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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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Project description:Cryptococcus neoformans strain H99 was exposed to 3μg/ml amphotericin B. Among the amphotericin B-tolerant adaptors, transcriptomes of four adaptors (TJ1844, TJ1849, TJ1852, TH1854) were analyzed and compared to wild type strain H99.

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TJ1843 derived amphotericin B (8ug/ml) adaptors

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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Project description:The gene expression of C. neoformans H99 treated with eltrombopag

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Different Pathways Mediate Amphotericin-Lactoferrin Drug Synergy in Cryptococcus and Saccharomyces.

Project description: Not available

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