Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Aspergillus fumigatus in blood reveals a "just wait and see" resting stage behavior

ABSTRACT: Objective: We analyzed changes in A. fumigatus gene expression profile at various stages of an in vitro model of aspergillosis to study the adaptation of A. fumigatus to the blood environment. Results: Most of virulence factors described to be involved in aspergillosis were not activated during the blood phase. We found three active processes to be activated in the later phase that may help to the adaptation: Iron homeostasis, a partial secondary metabolite cluster and the formation of detoxification enzymes. Conclusions: We propose that A. fumigatus is unable to grow in blood and it requires a metabolic change that allows the organism to shut down all uptake and energy-consume mechanisms, resulting in a resting mycelial stage. We performed gene expression profile by sequencing mRNA of A. fumigatus that were growm under two conditions, Minimal Medium (M) and human blood (B), and at different times: before placing the fungus in the final medium (pre), at 30' and at 180', with 2 biological replicates per condition.

ORGANISM(S): Aspergillus fumigatus

SUBMITTER: Sonia Tarazona

PROVIDER: E-GEOD-70227 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

RNAseq analysis of Aspergillus fumigatus in blood reveals a just wait and see resting stage behavior.

Irmer Henriette H Tarazona Sonia S Sasse Christoph C Olbermann Patrick P Loeffler Jürgen J Krappmann Sven S Conesa Ana A Braus Gerhard H GH

BMC genomics 20150827

<h4>Background</h4>Invasive aspergillosis is started after germination of Aspergillus fumigatus conidia that are inhaled by susceptible individuals. Fungal hyphae can grow in the lung through the epithelial tissue and disseminate hematogenously to invade into other organs. Low fungaemia indicates that fungal elements do not reside in the bloodstream for long.<h4>Results</h4>We analyzed whether blood represents a hostile environment to which the physiology of A. fumigatus has to adapt. An in vitr ...[more]

PMID: 26311470

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Project description:We examined the antifungal activity of artemisinin against Aspergillus fumigatus (A. fumigatus), a pathogenic filamentous fungus responsible for allergic and invasive aspergillosis in humans and analyzed transcript profiles of the fungus on exposure to Artemisinin. A. fumigatus spores were cultured for 48 h and then treated with artemisinin (at MIC50 concentration) or solvent control (DMSO) for 3 h to study its transcriptomic profiles. Total RNA (7 M-BM-5g) from A. fumigatus treated with solvent control (DMSO) or artemisinin was reverse transcribed separately using fluorescein-labeled dCTP or biotin-labeled dCTP from Micromax TSA labeling kit (Cy3 or Cy5 dyes are differentially deposited to these labeled cDNAs, post-hybridization, using tyramide signal amplification (TSA) protocol). Labeled cDNAs from test and control sample were mixed in equal amount and hybridized to microarray slide(s). After hybridization, the slides were processed as per the protocol of the Micromax TSA labeling kit and scanned in both the Cy3 and the Cy5 channels with a model 4200 Axon Instruments scanner with a 10-M-BM-5m resolution. Each of the signals was converted into a resolution of 16 bits per pixel. A dye swap experiment with a biological replicate was performed to ensure the reproducibility of each gene expression pattern.TIFF images were analyzed using GenePix Pro 6.0 software. The signal intensity value of each spot was reduced by the local background level for each spot. Diagnosis and normalization of microarray data (DNMAD) program was used for print-tip loess normalization of microarray data and for merging the data from dye-swap experiments . M-values obtained from this program were converted to log ratio (test/reference) and fold modulation. Genes having less than 3 spots in 2 dye swap slides were discarded and only those having their all fold change values (from 3 or 4 replicates in 2 slides; as each target is spotted twice on the slide) within a coefficient of variation M-BM-1 0.5 were used for analysis. Gene expression ratios of M-bM-^IM-%2 (fold upregulation or downregulation) in both replicates (dye swap) were considered to be differentially expressed genes.

Dataset Information

Aspergillus fumigatus in blood reveals a "just wait and see" resting stage behavior

Publications

RNAseq analysis of Aspergillus fumigatus in blood reveals a just wait and see resting stage behavior.

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