Natasa Radakovic1*, Ivana Moric1, Dejan Opsenica2,3 and Lidija Senerovic1
1Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Serbia
2Institute of Chemistry, Technology, and Metallurgy, University of Belgrade
3Centre of Excellence in Environmental Chemistry and Engineering, ICTM
natasa.radakovic [at] imgge.bg.ac.rs
Abstract
The ability of Candida albicans to undergo yeast-to-hypha transition is a key determinant of its virulence and represents an attractive target for anti-virulence strategies. In our previous work, we synthesized and characterized a candidate compound structurally related to 2-heptyl-4-quinolone (HHQ), which inhibits filamentation. Here, we aimed to elucidate its mechanism of action using transcriptomic profiling and functional validation.
RNA-seq analysis was performed in RPMI medium, a condition that supports filamentation, enabling assessment of compound effects on the hypha-associated transcriptional program. Differential gene expression analysis was conducted by comparing treated samples to untreated controls. In addition, the compound’s activity was compared to that of HHQ, a reference molecule from the same structural class.
Transcriptomic data revealed significant downregulation of hypha-associated genes, including ECE1 and ALS3, indicating suppression of the filamentation-associated transcriptional program. Functional validation using an ECE1 reporter strain confirmed a marked reduction in ECE1 expression, accompanied by inhibition of filamentation at the phenotypic level. Notably, both transcriptional and phenotypic effects of the tested compound were more pronounced than those observed with HHQ.
Together, these findings indicate that the compound exerts a targeted anti-virulence effect by suppressing key regulatory pathways associated with filamentation in C. albicans. This study highlights the value of integrating transcriptomic analysis with functional assays to characterize novel anti-virulence agents.
Keywords: Candida albicans, transcriptomics, ECE1, anti-virulence