Hsp90 governs echinocandin resistance in the pathogenic yeast Candida albicans via calcineurin.
Candida albicans is the leading fungal pathogen of humans, causing life-threatening disease in immunocompromised individuals. Treatment of candidiasis is hampered by the limited number of antifungal drugs whose efficacy is compromised by host toxicity, fungistatic activity, and the emergence of drug resistance. We previously established that the molecular chaperone Hsp90, which regulates the form and function of diverse client proteins, potentiates resistance to the azoles in C. albicans and in the model yeast Saccharomyces cerevisiae. Genetic studies in S. cerevisiae revealed that Hsp90's role in azole resistance is to enable crucial cellular responses to the membrane stress exerted by azoles via the client protein calcineurin. Here, we demonstrate that Hsp90 governs cellular circuitry required for resistance to the only new class of antifungals to reach the clinic in decades, the echinocandins, which inhibit biosynthesis of a critical component of the fungal cell wall. Pharmacological or genetic impairment of Hsp90 function reduced tolerance of C. albicans laboratory strains and resistance of clinical isolates to the echinocandins and created a fungicidal combination. Compromising calcineurin function phenocopied compromising Hsp90 function. We established that calcineurin is an Hsp90 client protein in C. albicans: reciprocal co-immunoprecipitation validated physical interaction; Hsp90 inhibition blocked calcineurin activation; and calcineurin levels were depleted upon genetic reduction of Hsp90. The downstream effector of calcineurin, Crz1, played a partial role in mediating calcineurin-dependent stress responses activated by echinocandins. Hsp90's role in echinocandin resistance has therapeutic potential given that genetic compromise of C. albicans HSP90 expression enhanced the efficacy of an echinocandin in a murine model of disseminated candidiasis. Our results identify the first Hsp90 client protein in C. albicans, establish an entirely new role for Hsp90 in mediating resistance to echinocandins, and demonstrate that targeting Hsp90 provides a promising therapeutic strategy for the treatment of life-threatening fungal disease.
Singh
Sheena D
SD
Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada.
Robbins
Nicole
N
Zaas
Aimee K
AK
Schell
Wiley A
WA
Perfect
John R
JR
Cowen
Leah E
LE
D000704:Analysis of Variance; D000818:Animals; D000935:Antifungal Agents / Q000494:pharmacology*; D001393:Azoles / Q000494:pharmacology; D019703:Calcineurin / Q000235:genetics / Q000378:metabolism*; D002176:Candida albicans / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D002177:Candidiasis / Q000188:drug therapy / Q000382:microbiology; D004268:DNA-Binding Proteins / Q000378:metabolism; D004195:Disease Models, Animal; D025141:Drug Resistance, Fungal; D054714:Echinocandins / Q000494:pharmacology*; D005656:Fungal Proteins / Q000378:metabolism; D018841:HSP90 Heat-Shock Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D055666:Lipopeptides / Q000494:pharmacology; D008297:Male; D000077551:Micafungin; D051379:Mice; D013312:Stress, Physiological; D014157:Transcription Factors / Q000378:metabolism
D000935:Antifungal Agents; D001393:Azoles; D004268:DNA-Binding Proteins; D054714:Echinocandins; D005656:Fungal Proteins; D018841:HSP90 Heat-Shock Proteins; D055666:Lipopeptides; D014157:Transcription Factors; D019703:Calcineurin; D000077551:Micafungin