Deletion of core septin gene aspB in Aspergillus fumigatus results in fungicidal activity of caspofungin.

Septins are a family of GTP-binding proteins found in many eukaryotic lineages. Although highly conserved throughout many eukaryotes, their functions vary across species. In Aspergillus fumigatus, the etiological agent of invasive aspergillosis, septins participate in a variety of processes such as cell wall organization of conidia, septation, and response to cell wall stress. Previous studies determined that the ΔaspB strain had a greater sensitivity to anti-cell wall drugs, especially the echinocandin caspofungin, yet mechanisms behind this augmented sensitivity are unknown. We performed cell viability staining of the deletion strains post-caspofungin exposure and found that the ΔaspA, ΔaspB, and ΔaspC strains have significantly lower cell viability. Concomitant with the reduced viability, deletion strains are more susceptible to caspofungin on solid media. These results indicate that the septin cytoskeleton is important for A. fumigatus survival in the presence of caspofungin. Due to the potential of improved therapeutic outcome, we followed up using a neutropenic murine model of invasive aspergillosis. Animals infected with the ΔaspB strain and treated with caspofungin showed improved survival compared to the animals infected with akuB KU80 wild-type or complemented strains. Additionally, histological analysis showed reduced fungal burden and inflammation in the ΔaspB infected, caspofungin-treated group. Affinity purification coupled with quantitative proteomics identified proteins involved in the septin-dependent response to caspofungin, includng four candidate interactors involved in cell wall stress response. Deletion of these candidate genes resulted in increased susceptibility to caspofungin and moderately reduced viability post-drug exposure. Taken together, these data suggest that septin AspB contributes to the fungistatic response to caspofungin.