Abstract
Aspergillus fumigatus is a saprobic fungus that causes a range of pulmonary diseases, some of which are characterised by fungal persistence such as is observed in cystic fibrosis (CF) patients. Creation of genetic variation is critical for A. fumigatus to adapt to the lung environment, but biofilm formation, especially in CF patients, may preclude mutational supply in A. fumigatus due to its confinement to the hyphal morphotype. We tested our hypothesis that genetic variation is created through parasexual recombination in chronic biofilms by phenotypic and genetic analysis of A. fumigatus isolates cultured from different origins.
As diploids are the hallmark of parasex, we screened 799 A. fumigatus isolates obtained from patients with CF, chronic pulmonary lung disease and acute invasive aspergillosis, and from the environment for spore size. Benomyl sensitivity, nuclear content measurements through fluorescence-activated cell sorting and scanning electron microscopy were used to confirm the diploid state of large size spores. Whole genome sequencing was used to characterise diploid-associated genetic variation.
We identified 11 diploids in isolates recovered from six of 11 (55%) CF patients and from one of 24 (4%) chronic aspergillosis patients, but not in 368 isolates from patients with acute Aspergillus infection and the environment. Diploid formation was associated with accumulation of mutations and variable haploid offspring including a voriconazole-resistant isolate.
Parasexual recombination allows A. fumigatus to adapt and persist in CF patients, and plays a role in azole resistance development. Our findings are highly significant for understanding the genetics and biology of A. fumigatus in the human lung.
Abstract
Aspergillus fumigatus can undergo parasexual recombination in the lungs of chronically colonised individuals. This is highly relevant, as the parasexual cycle can facilitate genetic variation and, consequently, in-host adaptation. https://bit.ly/3kAtrXd
Footnotes
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Conflict of interest: T. Engel has nothing to disclose.
Conflict of interest: P.E. Verweij reports grants from Gilead Sciences, MSD, Pfizer and F2G, and nonfinancial support from OLM and IMMY, outside the submitted work.
Conflict of interest: J. van den Heuvel has nothing to disclose.
Conflict of interest: D. Wangmo has nothing to disclose.
Conflict of interest: J. Zhang has nothing to disclose.
Conflict of interest: A.J.M. Debets has nothing to disclose.
Conflict of interest: E. Snelders has nothing to disclose.
- Received January 13, 2020.
- Accepted September 15, 2020.
- Copyright ©ERS 2020
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