Dataset Information

Serial passaging of Aspergillus fumigatus on Galleria extract agar produces strains with altered response of oxidative stress

ABSTRACT: Aspergillus fumigatus is an environmental saprophyte and opportunistic fungal pathogen of the human lung. Serially passaging of A. fumigatus on an agar produced from Galleria mellonella larvae produced strains showing alterations in virulence, anti-fungal susceptibility and in protein abundances that may indicate a degree of adaptation or selection after 25 passages on the artificial agar. While passaged strains demonstrated reduced virulence in G. mellonella larvae, they demonstrated increased tolerance to haemocyte mediated killing, hydrogen peroxide, itraconazole and amphotericin B. Label-free proteomic analysis of control and passaged A. fumigatus strains revealed a total of 3329 proteins of which, 1902 remained following filtration and 32 proteins were statistically significant and differentially abundant. Proteins that play a role in the oxidative stress response were altered in abundance in the passaged strain and included (S)-S-oxide reductase (+2.63 fold), developmental regulator FlbA (+2.27 fold) and histone H2A.Z (-1.82 fold). The results presented here indicate that prolonged culturing of A. fumigatus on Galleria extract agar results in alterations in susceptibility to oxidative stress and in the abundance of proteins associated with the oxidative stress response. The phenomenon may be a result of selection for strains suitable for survival in the adverse conditions imposed by the innate immune response.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Aspergillus Fumigatus Af293

SUBMITTER: Aaron Curtis

LAB HEAD: Professor Kevin Kavanagh

PROVIDER: PXD036787 | Pride | 2023-05-10

REPOSITORIES: Pride

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Prolonged Subculturing of Aspergillus fumigatus on Galleria Extract Agar Results in Altered Virulence and Sensitivity to Antifungal Agents.

Curtis Aaron A Walshe Kieran K Kavanagh Kevin K

Cells 20230331 7

Aspergillus fumigatus is an environmental saprophyte and opportunistic fungal pathogen of humans. The aim of the work presented here was to examine the effect of serially subculturing A. fumigatus on agar generated from Galleria mellonella larvae in order to characterize the alterations in the phenotypes that might occur. The passaged strains showed alterations in virulence, antifungal susceptibility, and in protein abundances that may indicate adaptation after 25 passages o ...[more]

PMID: 37048138

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Project description:Aspergillus fumigatus is an opportunistic fungal pathogen of the lungs resulting in both chronic and acute infection. The fungus interacts with numerous bacteria that compose the microflora of the lung including Pseudomonas aeruginosa and Klebsiella pneumoniae, both common isolates from Cystic fibrosis sputum. It is well documented that bacterial and fungal interactions both physically and through production of secreted products which can alter the growth and virulence of other species. The interaction with P. aeruginosa has been characterised previously indicating decreased growth and increased gliotoxin production in the presence of cells and the opposite when only exposed to culture filtrate. In our analysis exposure of cell free culture filtrate inhibited fungal growth and increased gliotoxin production. This was supported with proteomic analysis indicating reduction in 1,3-beta-glucanosyltransferase (-3.97 fold), methyl sterol monooxygenase erg25B (-2.9 fold) and Calcium/calmodulin dependent protein kinase (-4.2 fold) involved in fungal development. Coupled with increased expression of Glutathione S-transferase gliG (6.17 fold), non-ribosomal peptide synthase gliP (3.67 fold), O-methyltransferase gliM (3.5 fold), Gamma-glutamyl acyltransferase gliK (2.89 fold) and thioredoxin reductase gliT (2.33 fold) involved in Gliotoxin production indicate a possible shift towards secondary metabolism. Qualitative proteomic analysis of the bacterial supernatant also elucidates possible mechanisms for these alterations including physical interaction by Outer membrane protein A, High-affinity zinc uptake system protein ZnuA and Iron uptake system component EfeO reducing essential nutrient availability, enzymatic activity including Periplasmic serine endoprotease DegP-like and redox active proteins including Alkyl hydroperoxide reductase C. These proteomic alterations to the fungus could have implications for the severity of virulence in the context of coinfection and may impact disease progression.

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Dataset Information

Serial passaging of Aspergillus fumigatus on Galleria extract agar produces strains with altered response of oxidative stress

Dataset's files

Publications

Prolonged Subculturing of <i>Aspergillus fumigatus</i> on <i>Galleria</i> Extract Agar Results in Altered Virulence and Sensitivity to Antifungal Agents.

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