Characterization of echinocandin-resistant mutants of Candida albicans: genetic, biochemical, and virulence studies.
The pneumocandins are potent antifungal agents of the echinocandin class which are under development for use as broad-spectrum antimycotic therapy. One important consideration for any new therapeutic class for treating serious fungal infections is the potential for drug resistance development. In this study we have isolated and characterized four independent spontaneous Candida albicans mutants resistant to the potent semisynthetic pneumocandin L-733,560. These mutants have many of the properties of FKS1/ETG1 echinocandin-resistant mutants of Saccharomyces cerevisiae, including (i) cross-resistance to other 1,3-beta-D-glucan synthase inhibitors, such as papulacandin and echinocandins, but no change in sensitivity to other antifungal agents; (ii) in vitro glucan synthase activity that is more resistant to pneumocandins than the wild-type parent enzyme; and (iii) semidominant drug resistance in spheroplast fusion strains. The mutants were compared with C. albicans echinocandin-resistant mutants isolated by mutagenesis by L. Beckford and D. Kerridge (mutant M-2) (abstr. PS3.11, in Proceedings of the XI Congress of the International Society for Human and Animal Mycology, Montreal, Canada, 1992) and by A. Cassone, R. E. Mason, and D. Kerridge (mutant CA-2) (Sabouraudia 19:97-110, 1981). All of the strains had resistant enzyme activity in vitro. M-2 grew poorly and had low levels of enzyme activity. In contrast, CA-2 and the spontaneous mutants grew as well as the parents and had normal levels of glucan synthase activity. These results suggest that these resistant mutants may have alterations in glucan synthase. CA-2 was unable to form germ tubes, an ability retained by the spontaneous mutants. The virulence of the spontaneous mutants was unimpaired in a mouse model of disseminated candidiasis, while M-2 and CA-2 were 2 orders of magnitude less virulent than their parent strains. Significantly, mice challenged with the spontaneous mutant CAI4R1 responded therapeutically to lower levels of L-733,560 than would he predicted by the increase in in vitro susceptibility.
Kurtz
M B
MB
Department of Infectious Disease Research, Merck Research Laboratories, Rahway, New Jersey 07065, USA.
Abruzzo
G
G
Flattery
A
A
Bartizal
K
K
Marrinan
J A
JA
Li
W
W
Milligan
J
J
Nollstadt
K
K
Douglas
C M
CM
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D000935:Antifungal Agents / Q000494:pharmacology*; D002176:Candida albicans / Q000235:genetics* / Q000472:pathogenicity; D004305:Dose-Response Relationship, Drug; D004352:Drug Resistance, Microbial / Q000235:genetics; D054714:Echinocandins; D005260:Female; D005656:Fungal Proteins; D005799:Genes, Dominant; D005964:Glucosyltransferases / Q000037:antagonists & inhibitors; D007928:Lethal Dose 50; D051379:Mice; D008811:Mice, Inbred DBA; D008826:Microbial Sensitivity Tests; D009154:Mutation; D010455:Peptides; D010456:Peptides, Cyclic / Q000494:pharmacology; D011714:Pyrans / Q000494:pharmacology; D013104:Spheroplasts / Q000187:drug effects; D014774:Virulence / Q000235:genetics
D000900:Anti-Bacterial Agents; D000935:Antifungal Agents; D054714:Echinocandins; D005656:Fungal Proteins; C089367:L 733560; D010455:Peptides; D010456:Peptides, Cyclic; D011714:Pyrans; C065110:L 687781; C079465:pneumocandin B(0); C007572:echinocandin B; D005964:Glucosyltransferases; C040232:glucan synthase