Invasive fungal infections due to Candida species are a major cause of morbidity and mortality. Timely diagnosis and appropriate antifungal therapy are critical for patient management. Unfortunately, there are relatively few drug classes available for therapy and they are being compromised by drug resistance. The echinocandins are important antifungal agents for the treatment of patients with Candida infections. However, therapeutic failures are increasingly being reported especially with C. glabrata, which is a leading cause of disseminated candidiasis. Of the major Candida species, echinocandin resistance remains relatively uncommon. Yet, this is not the case for C. glabrata, where resistance now exceeds 13% in certain high-risk centers. It is well- established that clinical echinocandin drug resistance resulting in therapeutic failure is due to amino acid substitutions in highly conserved hot-spot regions of the Fks subunits of glucan synthase. There is a critical need to understand the cellular factors underlying the emergence of Fks-mediated echinocandin resistance. The novel hypothesis being explored in this proposal is that echinocandin resistance in C. glabrata is promoted by cellular factors that stabilize cells in response to drug by creating an unanticipated drug tolerant cell population, which ultimately escapes drug action by the formation of characteristic fks mutations. This hypothesis is supported by pharmacodynamic studies in mice indicating that echinocandin drug therapy at a standard dosage (human equivalent) results in drug tolerant or persistent cell populations. Such behavior is inconsistent with echinocandin drugs as presumed fungicidal agents. Rather, the in vivo data suggests that the echinocandin drugs behave as fungistatic agents at a standard dosage. It is hypothesized that in vivo cells become temporally drug- tolerant by inducing a variety of cellular compensatory mechanisms in response to cell wall stress due to echinocandin action. Cellular and genetic factors promote selection of drug resistant variants with mutations in FKS genes from high burden populations of drug tolerant persistor cells. To better understand echinocandin resistance in C. glabrata, a range of cellular factors will be examined in in vitro and in vivo models for their role in emergence of Fks-mediated echinocandin resistance. Specifically, the importance of compensatory cell wall stress responses, decreased DNA repair, azole resistance, serum, and potential novel genes/pathways will be evaluated using appropriate mutants, along with known inhibitors of these pathways. This work will exploit engineered isogenic mutant strains and a unique collection of serial genetically-matched susceptible and fks-resistant clinical isolates of C. glabrata. These latter isolates will be profiled for changes in the genome and transcriptome for the first time to assess the importance of known mechanisms and elucidate potential new genetic incites underlying resistance emergence. Our detailed molecular studies of echinocandin resistance with clinical isolates over the past decade provide a strong foundation to better understand the molecular basis underlying the emergence of echinocandin resistance in C. glabrata. It is anticipated that this information will provide important new insights and potential intervention strategies to overcome and/or prevent the emergence of echinocandin resistance.

Public Health Relevance

Invasive fungal infections are a significant cause of morbidity and mortality in immunocompromised patients, and treatment options are limited. The echinocandins are an important antifungal class for primary therapy but emerging resistance threatens their effectiveness. A greater understanding of factors contributing to resistance emergence is important to develop new strategies to overcome and/or prevent drug resistance.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
1R01AI109025-01
Application #
8614663
Study Section
Drug Discovery and Mechanisms of Antimicrobial Resistance Study Section (DDR)
Program Officer
Duncan, Rory A
Project Start
Project End
Budget Start
Budget End
Support Year
1
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Rutgers University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
City
Newark
State
NJ
Country
United States
Zip Code
07103
Arendrup, Maiken C; Perlin, David S (2014) Echinocandin resistance: an emerging clinical problem? Curr Opin Infect Dis 27:484-92
Perlin, David S (2014) Echinocandin resistance, susceptibility testing and prophylaxis: implications for patient management. Drugs 74:1573-85