Oliveira, N. K.; Bhattacharya, S.; Gambhir, R.; Joshi, M.; Fries, B. C. Novel ABC Transporter Associated with Fluconazole Resistance in Aging of Cryptococcus Neoformans. Journal of Fungi, 2022, 8, 677. https://doi.org/10.3390/jof8070677.
Oliveira, N. K.; Bhattacharya, S.; Gambhir, R.; Joshi, M.; Fries, B. C. Novel ABC Transporter Associated with Fluconazole Resistance in Aging of Cryptococcus Neoformans. Journal of Fungi, 2022, 8, 677. https://doi.org/10.3390/jof8070677.
Oliveira, N. K.; Bhattacharya, S.; Gambhir, R.; Joshi, M.; Fries, B. C. Novel ABC Transporter Associated with Fluconazole Resistance in Aging of Cryptococcus Neoformans. Journal of Fungi, 2022, 8, 677. https://doi.org/10.3390/jof8070677.
Oliveira, N. K.; Bhattacharya, S.; Gambhir, R.; Joshi, M.; Fries, B. C. Novel ABC Transporter Associated with Fluconazole Resistance in Aging of Cryptococcus Neoformans. Journal of Fungi, 2022, 8, 677. https://doi.org/10.3390/jof8070677.
Abstract
Cryptococcus neoformans causes meningoencephalitis in immunocompromised individuals, which is treated with Fluconazole (FLC) monotherapy when resources are limited. This can lead to azole resistance, which can be mediated by overexpression of ABC transporters, a class of efflux pumps. ABC pump-mediated efflux of FLC is also augmented in 10-generation old C. neoformans cells. Here, we describe a new ABC transporter Afr3 (CNAG_06909), which is overexpressed in C. neoformans cells of advanced generational age, that accumulate during chronic infection. The delta-afr3 mutant strain showed higher FLC susceptibility by FLC E-Test strip testing and also by a killing test that measured survival after 3 h FLC exposure. Furthermore, delta-afr3 cells exhibited lower Rhodamine 6G efflux compared to the H99 wild type cells. Afr3 was expressed in the Saccharomyces cerevisiae AD-delta strain, which lacks several drug transporters, thus reducing background transport. The AD-delta + Afr3 strain demonstrated a higher efflux with both Rhodamine 6G and Nile Red, even though the FLC MICs were not changed. Characterization of the delta-afr3 mutant revealed unattenuated growth but a prolongation (22%) of the replicative life span. In addition, delta-afr3 exhibited decreased resistance to macrophage killing and attenuated virulence in the Galleria mellonella infection model. In summary, our data indicate that a novel ABC pump Afr3p, which is upregulated in C. neoformans cells of advanced age may contribute to their enhanced FLC tolerance, by promoting drug efflux. Lastly, its role in macrophage resistance may also contribute to the selection of older C. neoformans cells during chronic infection.
Keywords
drug resistance; ABC transporter; efflux pump; cryptococcosis; aging
Subject
Biology and Life Sciences, Immunology and Microbiology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
