Use of Haploid Model of Candida albicans to Uncover Mechanism of Action of a Novel Antifungal Agent

Frontiers in Cellular and Infection Microbiology

Yue Wang¹  Guisheng Zeng²  Chaminda J. Seneviratne³  Tanujaa Suriyanarayanan³  Thuyen Truong³  Cao Tong³  Thuc D. Le⁴  Jiuyong Li⁴  Lin Liu⁴ 

[1] Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore;Institute of Molecular and Cell Biology, Agency for Science, Technology and Research, Singapore, Singapore;Oral Sciences, Faculty of Dentistry, National University of Singapore, Singapore, Singapore;School of Information Technology and Mathematical Sciences, University of South Australia, Adelaide, SA, Australia;

关键词: Candida albicans;    haploid;    SM21;    mechanism of action;    mitochondria;   

DOI  :  10.3389/fcimb.2018.00164

来源: DOAJ

【 摘 要 】

Antifungal agents for the treatment of Candida albicans infections are limited. We recently discovered a novel antifungal small molecule, SM21, with promising in vivo activity. Herein, we employed the newly developed C. albicans haploid toolbox to uncover the mechanism of action of SM21. Comprehensive RNA-Seq analyses of the haploid susceptible GZY803 strain revealed significant gene expression changes related to mitochondria when exposed to SM21. Mitochondrial structure visualization and measurement of ATP generation, reactive oxygen species (ROS) levels, and the antioxidant potential of SM21-treated and untreated GZY803, mitochondrial structure defective haploid mutant (dnm1Δ), and wild-type diploid SC5314 strains confirmed defects in mitochondria. Exploiting the advantage of C. albicans haploids as a single ploidy model, we further exposed GZY803 to repetitive treatments of SM21 in order to generate resistant mutants. Three colonies designated S3, S5 and S6, which displayed resistance to SM21, were isolated. All resistant strains exhibited enhanced transcriptomic responses for peptide and protein metabolism and secreted aspartate proteases (SAPs) activity under SM21 treatment compared to the parent strain GZY803. Consistently, supplementing the resistant strains, GZY803, and SC5314 with peptone, a form of digested peptides, decreased susceptibility to SM21. The present study demonstrates the usefulness of haploid C. albicans model in antifungal drug discovery. The findings will be invaluable to develop SM21 as a novel antifungal agent, which will benefit millions of patients suffering from Candida infections.

【 授权许可】

Unknown