title: Novel high-throughput screening method for identification of fungal dimorphism blockers.
abstract: Invasive mycoses have been increasing worldwide, with Candida spp. being the most prevalent fungal pathogen causing high morbidity and mortality in immunocompromised individuals. Only few antimycotics exist, often with severe side effects. Therefore, new antifungal drugs are urgently needed. Because the identification of antifungal compounds depends on fast and reliable assays, we present a new approach based on high-throughput image analysis to define cell morphology. Candida albicans and other fungi of the Candida clade switch between different growth morphologies, from budding yeast to filamentous hyphae. Yeasts are considered proliferative, whereas hyphae are required for invasion and dissemination. Thus, morphotype switching in many Candida spp. is connected to virulence and pathogenesis. It is, consequently, reasonable to presume that morphotype blockers interfere with the virulence, thereby preventing hazardous colonization. Our method efficiently differentiates yeast from hyphal cells using a combination of automated microscopy and image analysis. We selected the parameters length/width ratio and mean object shape to quantitatively discriminate yeasts and hyphae. Notably, Z' factor calculations for these parameters confirmed the suitability of our method for high-throughput screening. As a second stage, we determined cell viability to discriminate morphotype-switching inhibitors from those that are fungicidal. Thus, our method serves as a basis for the identification of candidates for next-generation antimycotics.
authors.affiliation: Department of Clinical Microbiology, Umeå University, Umeå, Sweden Umeå Centre for Microbial Research (UCMR), Umeå, Sweden Laboratory for Infection Medicine Sweden (MIMS), Umeå University, Umeå, Sweden.
authors.forename: Marios
authors.initials: M
authors.lastname: Stylianou
authors.affiliation: Umeå Centre for Microbial Research (UCMR), Umeå, Sweden Department of Chemistry, Umeå University, Umeå, Sweden.
authors.forename: Hanna
authors.initials: H
authors.lastname: Uvell
authors.affiliation: Department of Clinical Microbiology, Umeå University, Umeå, Sweden Umeå Centre for Microbial Research (UCMR), Umeå, Sweden Laboratory for Infection Medicine Sweden (MIMS), Umeå University, Umeå, Sweden.
authors.forename: José Pedro
authors.initials: JP
authors.lastname: Lopes
authors.affiliation: Umeå Centre for Microbial Research (UCMR), Umeå, Sweden Department of Chemistry, Umeå University, Umeå, Sweden.
authors.forename: Per-Anders
authors.initials: PA
authors.lastname: Enquist
authors.affiliation: Umeå Centre for Microbial Research (UCMR), Umeå, Sweden Department of Chemistry, Umeå University, Umeå, Sweden.
authors.forename: Mikael
authors.initials: M
authors.lastname: Elofsson
authors.affiliation: Department of Clinical Microbiology, Umeå University, Umeå, Sweden Umeå Centre for Microbial Research (UCMR), Umeå, Sweden Laboratory for Infection Medicine Sweden (MIMS), Umeå University, Umeå, Sweden constantin.urban@umu.se.
authors.forename: Constantin F
authors.initials: CF
authors.lastname: Urban
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000935:Antifungal Agents / Q000494:pharmacology*; D002175:Candida / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D025301:Hyphae / Q000187:drug effects*; D008826:Microbial Sensitivity Tests / Q000379:methods; D050296:Microbial Viability / Q000187:drug effects; D008853:Microscopy / Q000379:methods
chemical_list: D000935:Antifungal Agents; D000255:Adenosine Triphosphate
keywords: Candida; antifungal compounds; high-throughput screening; mycoses; yeast-to-hypha transition