Abstract Description

The publishing of the World Health Organisation's first Fungal Priority Pathogen List (WHO FPPL) critical group emphasises the significant level of public health impact of fungal pathogens and the need to develop new and improved diagnostic tools, treatments, and preventive measures for these pathogens. Extracellular vesicles (EVs) are nanoparticles composed of lipid bilayered membrane structures, considered "virulence bags," which carry several biomolecules, including proteins and lipids. Depending on the fungal pathogen, EVs can interact with the host immune system to elicit multiple outcomes and exacerbate or attenuate fungal infections. Here, focus on Candida albicans and its lipid metabolism-related gene, EHT1. We employed a CRISPR-Cas9 system to construct a homozygous mutant of EHT1 and performed RNA sequence analysis to identify the differentially expressed genes in mutant vs. wild-type biofilms. Furthermore, we evaluated the effect of the deletion on C. albicans growth, response to stress, hyphal formation, and, importantly, EV production and cargo. Our results show an overlap of the whole cell transcriptomics and EV protein cargo, emphasising that the content of EVs represents the metabolic state of the cells from which they were derived. In addition, our multi-omics results indicate that genes involved in lipid metabolism play a role in key virulence factors such as biofilm hyphal formation and cell wall organisation and EV production in C. albicans. These findings provide insight into biofilm pathogenic mechanisms and opportunities for the development of antifungal therapies.