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

Costa C, Ribeiro J, Miranda IM, Silva-Dias A, Cavalheiro M, Costa-de-Oliveira S, Rodrigues AG, Teixeira MC - Front Microbiol (2016)

Bottom Line: For years, antifungal drug resistance in Candida species has been associated to the expression of ATP-Binding Cassette (ABC) multidrug transporters.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.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Bioengineering, Instituto Superior Técnico, University of LisbonLisboa, Portugal; Institute for Bioengineering and Biosciences, Biological Sciences Research GroupLisboa, Portugal.

ABSTRACT
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 efflux 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.

No MeSH data available.


Transcript levels of CgAQR1, CgQDR2, CgTPO1_1, CgTPO1_2, CgTPO3, CgCDR1, and CgCDR2 in the C. glabrata clinical isolate 51800, when compared to those determined in the susceptible isolate exhibiting the lowest level of expression for each of these genes. Transcript levels were assessed through quantitative RT-PCR, as described in the section “Materials and Methods.” The obtained values are the average of at least three independent experiments. Error bars represent the corresponding standard deviation.
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Figure 5: Transcript levels of CgAQR1, CgQDR2, CgTPO1_1, CgTPO1_2, CgTPO3, CgCDR1, and CgCDR2 in the C. glabrata clinical isolate 51800, when compared to those determined in the susceptible isolate exhibiting the lowest level of expression for each of these genes. Transcript levels were assessed through quantitative RT-PCR, as described in the section “Materials and Methods.” The obtained values are the average of at least three independent experiments. Error bars represent the corresponding standard deviation.

Mentions: Based on the obtained susceptibility results, it appears clear that CgTpo3 is a player in clinical drug resistance acquisition, but is not fully responsible for the resistance to clotrimazole and fluconazole in the 51800 isolate. Indeed, the expression of additional multidrug transporter encoding genes in this strain was found to be quite high, when compared to what was determined in azole susceptible dose-dependent isolates (Figure 5). This is particularly the case for the CDR1 and AQR1 genes (Figure 4), highlighting the multifactorial nature of azole drug resistance acquisition in the clinical setting.


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

Costa C, Ribeiro J, Miranda IM, Silva-Dias A, Cavalheiro M, Costa-de-Oliveira S, Rodrigues AG, Teixeira MC - Front Microbiol (2016)

Transcript levels of CgAQR1, CgQDR2, CgTPO1_1, CgTPO1_2, CgTPO3, CgCDR1, and CgCDR2 in the C. glabrata clinical isolate 51800, when compared to those determined in the susceptible isolate exhibiting the lowest level of expression for each of these genes. Transcript levels were assessed through quantitative RT-PCR, as described in the section “Materials and Methods.” The obtained values are the average of at least three independent experiments. Error bars represent the corresponding standard deviation.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4835504&req=5

Figure 5: Transcript levels of CgAQR1, CgQDR2, CgTPO1_1, CgTPO1_2, CgTPO3, CgCDR1, and CgCDR2 in the C. glabrata clinical isolate 51800, when compared to those determined in the susceptible isolate exhibiting the lowest level of expression for each of these genes. Transcript levels were assessed through quantitative RT-PCR, as described in the section “Materials and Methods.” The obtained values are the average of at least three independent experiments. Error bars represent the corresponding standard deviation.
Mentions: Based on the obtained susceptibility results, it appears clear that CgTpo3 is a player in clinical drug resistance acquisition, but is not fully responsible for the resistance to clotrimazole and fluconazole in the 51800 isolate. Indeed, the expression of additional multidrug transporter encoding genes in this strain was found to be quite high, when compared to what was determined in azole susceptible dose-dependent isolates (Figure 5). This is particularly the case for the CDR1 and AQR1 genes (Figure 4), highlighting the multifactorial nature of azole drug resistance acquisition in the clinical setting.

Bottom Line: For years, antifungal drug resistance in Candida species has been associated to the expression of ATP-Binding Cassette (ABC) multidrug transporters.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.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Bioengineering, Instituto Superior Técnico, University of LisbonLisboa, Portugal; Institute for Bioengineering and Biosciences, Biological Sciences Research GroupLisboa, Portugal.

ABSTRACT
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 efflux 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.

No MeSH data available.