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For years, antifungal drug resistance in  Candida  species has been associated to the expression of ATP-Binding Cassette (ABC) multidrug transporters. More recently, a few drug eux pumps from the Drug:H +  Antiporter (DHA) family have also been shown to play a role in this process, although to date only the  Candida albicans  Mdr1 transporter has been demonstrated to be relevant in the clinical acquisition of antifungal drug resistance. This work provides evidence to suggest the involvement of the  C. glabrata  DHA transporters CgAqr1, CgQdr2, CgTpo1_1, and CgTpo3 in the clinical acquisition of clotrimazole drug resistance. A screening for azole drug resistance in 138  C. glabrata  clinical isolates, from patients attending two major Hospitals in Portugal, was performed. Based on this screening, 10 clotrimazole susceptible and 10 clotrimazole resistant isolates were selected for further analysis. The transcript levels of  CgAQR1 ,  CgQDR2 ,  CgTPO1_1 , and  CgTPO3  were found to be significantly up-regulated in resistant isolates when compared to the susceptible ones, with a level of correlation that was found to be similar to that of  CgCDR2 , an ABC gene known to be involved in the clinical acquisition of resistance. As a proof-of-concept experiment, the  CgTPO3  gene was deleted in an azole resistant  C. glabrata  isolate, exhibiting high levels of expression of this gene. The deletion of  CgTPO3  in this isolate was found to lead to decreased resistance to clotrimazole and fluconazole, and increased accumulation of azole drugs, thus suggesting the involvement of this transporter in the manifestation of azole resistance.

frontiers in microbiology

Clotrimazole Drug Resistance in Candida glabrata Clinical Isolates Correlates with Increased Expression of the Drug:H+ Antiporters CgAqr1, CgTpo1_1, CgTpo3, and CgQdr2