Extracellular DNA release confers heterogeneity in Candida albicans biofilm formation

Ranjith Rajendran, Leighann Sherry, David F Lappin, Chris J Nile, Karen Smith, Craig Williams, Carol A Munro, Gordon Ramage

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

BACKGROUND: Biofilm formation by Candida albicans has shown to be highly variable and is directly associated with pathogenicity and poor clinical outcomes in patients at risk. The aim of this study was to test the hypotheses that the extracellular DNA release by C. albicans is strain dependent and is associated with biofilm heterogeneity.

RESULTS: Initially, biofilm formed by C. albicans high biofilm formers (HBF) or low biofilm formers (LBF) were treated with DNase to find whether eDNA play a role in their biofilm formation. Digestion of biofilm eDNA significantly reduced the HBF biofilm biomass by five fold compared to untreated controls. In addition, quantification of eDNA over the period of biofilm formation by SYBR green assay demonstrate a significantly higher level of 2 to 6 fold in HBF compared to LBF. Biochemical and transcriptional analyses showed that chitinase activity and mRNA levels of chitinase genes, a marker of autolysis, were upregulated in 24 h biofilm formation by HBF compared to LBF, indicating autolysis pathway possibly involved in causing variation. The biofilm biomass and eDNA release by single (∆cht2, ∆cht3) and double knockout (∆cht2/∆cht3) chitinase mutants were significantly less compared to their parental strain CA14, confirming the role of chitinases in eDNA release and biofilm formation. Correlation analysis found a positive correlation between chitinases and HWP1, suggesting eDNA may release during the hyphal growth. Finally, we showed a combinational treatment of biofilms with DNase or chitinase inhibitor (acetazolamide) plus amphotericin B significantly improved antifungal susceptibility by 2 to 8 fold.

CONCLUSIONS: Collectively, these data show that eDNA release by C. albicans clinical isolates is variable and is associated with differential biofilm formation. Digestion of biofilm eDNA by DNase may provide a novel therapeutic strategies to destabilise biofilm growth and improves antifungal sensitivity.

Original languageEnglish
Pages (from-to)303
JournalBMC Microbiology
Volume14
DOIs
Publication statusPublished - 5 Dec 2014

Fingerprint

Biofilms
Candida albicans
DNA
Chitinases
Deoxyribonucleases
Autolysis
Biomass
Digestion
Acetazolamide

Keywords

  • Biofilms
  • Candida albicans
  • Chitinase
  • DNA, Fungal
  • Gene Expression Profiling
  • Humans
  • RNA, Messenger

Cite this

Rajendran, Ranjith ; Sherry, Leighann ; Lappin, David F ; Nile, Chris J ; Smith, Karen ; Williams, Craig ; Munro, Carol A ; Ramage, Gordon. / Extracellular DNA release confers heterogeneity in Candida albicans biofilm formation. In: BMC Microbiology. 2014 ; Vol. 14. pp. 303.
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Rajendran, R, Sherry, L, Lappin, DF, Nile, CJ, Smith, K, Williams, C, Munro, CA & Ramage, G 2014, 'Extracellular DNA release confers heterogeneity in Candida albicans biofilm formation', BMC Microbiology, vol. 14, pp. 303. https://doi.org/10.1186/s12866-014-0303-6

Extracellular DNA release confers heterogeneity in Candida albicans biofilm formation. / Rajendran, Ranjith; Sherry, Leighann; Lappin, David F; Nile, Chris J; Smith, Karen; Williams, Craig; Munro, Carol A; Ramage, Gordon.

In: BMC Microbiology, Vol. 14, 05.12.2014, p. 303.

Research output: Contribution to journalArticle

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T1 - Extracellular DNA release confers heterogeneity in Candida albicans biofilm formation

AU - Rajendran, Ranjith

AU - Sherry, Leighann

AU - Lappin, David F

AU - Nile, Chris J

AU - Smith, Karen

AU - Williams, Craig

AU - Munro, Carol A

AU - Ramage, Gordon

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N2 - BACKGROUND: Biofilm formation by Candida albicans has shown to be highly variable and is directly associated with pathogenicity and poor clinical outcomes in patients at risk. The aim of this study was to test the hypotheses that the extracellular DNA release by C. albicans is strain dependent and is associated with biofilm heterogeneity.RESULTS: Initially, biofilm formed by C. albicans high biofilm formers (HBF) or low biofilm formers (LBF) were treated with DNase to find whether eDNA play a role in their biofilm formation. Digestion of biofilm eDNA significantly reduced the HBF biofilm biomass by five fold compared to untreated controls. In addition, quantification of eDNA over the period of biofilm formation by SYBR green assay demonstrate a significantly higher level of 2 to 6 fold in HBF compared to LBF. Biochemical and transcriptional analyses showed that chitinase activity and mRNA levels of chitinase genes, a marker of autolysis, were upregulated in 24 h biofilm formation by HBF compared to LBF, indicating autolysis pathway possibly involved in causing variation. The biofilm biomass and eDNA release by single (∆cht2, ∆cht3) and double knockout (∆cht2/∆cht3) chitinase mutants were significantly less compared to their parental strain CA14, confirming the role of chitinases in eDNA release and biofilm formation. Correlation analysis found a positive correlation between chitinases and HWP1, suggesting eDNA may release during the hyphal growth. Finally, we showed a combinational treatment of biofilms with DNase or chitinase inhibitor (acetazolamide) plus amphotericin B significantly improved antifungal susceptibility by 2 to 8 fold.CONCLUSIONS: Collectively, these data show that eDNA release by C. albicans clinical isolates is variable and is associated with differential biofilm formation. Digestion of biofilm eDNA by DNase may provide a novel therapeutic strategies to destabilise biofilm growth and improves antifungal sensitivity.

AB - BACKGROUND: Biofilm formation by Candida albicans has shown to be highly variable and is directly associated with pathogenicity and poor clinical outcomes in patients at risk. The aim of this study was to test the hypotheses that the extracellular DNA release by C. albicans is strain dependent and is associated with biofilm heterogeneity.RESULTS: Initially, biofilm formed by C. albicans high biofilm formers (HBF) or low biofilm formers (LBF) were treated with DNase to find whether eDNA play a role in their biofilm formation. Digestion of biofilm eDNA significantly reduced the HBF biofilm biomass by five fold compared to untreated controls. In addition, quantification of eDNA over the period of biofilm formation by SYBR green assay demonstrate a significantly higher level of 2 to 6 fold in HBF compared to LBF. Biochemical and transcriptional analyses showed that chitinase activity and mRNA levels of chitinase genes, a marker of autolysis, were upregulated in 24 h biofilm formation by HBF compared to LBF, indicating autolysis pathway possibly involved in causing variation. The biofilm biomass and eDNA release by single (∆cht2, ∆cht3) and double knockout (∆cht2/∆cht3) chitinase mutants were significantly less compared to their parental strain CA14, confirming the role of chitinases in eDNA release and biofilm formation. Correlation analysis found a positive correlation between chitinases and HWP1, suggesting eDNA may release during the hyphal growth. Finally, we showed a combinational treatment of biofilms with DNase or chitinase inhibitor (acetazolamide) plus amphotericin B significantly improved antifungal susceptibility by 2 to 8 fold.CONCLUSIONS: Collectively, these data show that eDNA release by C. albicans clinical isolates is variable and is associated with differential biofilm formation. Digestion of biofilm eDNA by DNase may provide a novel therapeutic strategies to destabilise biofilm growth and improves antifungal sensitivity.

KW - Biofilms

KW - Candida albicans

KW - Chitinase

KW - DNA, Fungal

KW - Gene Expression Profiling

KW - Humans

KW - RNA, Messenger

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