Development of an Imaging Flow Cytometry Method for Fungal Cytological Profiling and Its Potential Application in Antifungal Drug Development.

Automated imaging techniques have been in increasing demand for the more advanced analysis and efficient characterization of cellular phenotypes. The success of the image-based profiling method hinges on assays that can rapidly and simultaneously capture a wide range of phenotypic features. We have developed an automated image acquisition method for fungal cytological profiling (FCP) using an imaging flow cytometer that can objectively measure over 250 features of a single fungal cell.

Characterization of a Biaryl Amide Anti-virulence Compound Targeting Filamentation and Biofilm Formation.

We have previously identified a small molecule compound, N-[3-(allyloxy)-phenyl]-4-methoxybenzamide (9029936), that exerts potent inhibitory activity against filamentation and biofilm formation by the SC5314 strain and represents a lead candidate for the development of anti-virulence approaches against infections. Here we present data from a series of experiments to further characterize its activity and drug-like characteristics. We demonstrate the activity of this compound against a panel of clinical isolates, including several displaying resistance to current antifungals; as well as against a set of gain of function strains in key transcriptional regulators of antifungal drug resistance.

Systematic Complex Haploinsufficiency-Based Genetic Analysis of Transcription Factors: Tools and Applications to Virulence-Associated Phenotypes.

Genetic interaction analysis is a powerful approach to the study of complex biological processes that are dependent on multiple genes. Because of the largely diploid nature of the human fungal pathogen , genetic interaction analysis has been limited to a small number of large-scale screens and a handful for gene-by-gene studies. Complex haploinsufficiency, which occurs when a strain containing two heterozygous mutations at distinct loci shows a phenotype that is distinct from either of the corresponding single heterozygous mutants, is an expedient approach to genetic interactions analysis in diploid organisms.