Combatting fungal infections through the discovery and elucidation of novel anti‐virulence strategies

Abstract

Fungal pathogens are critically important threats to global health with over 300 million people affected by serious fungal diseases worldwide. Fungal pathogens have evolved sophisticated strategies, including the secretion of virulence factors to interfere with host cell functions and to perturb immune responses. Our ‘infectome’ analysis identifies previously undescribed proteins involved in fungal virulence and host immune response, representing an opportunity to elucidate molecular mechanisms of host-pathogen interplay during disease. Using state-of-the-art mass spectrometry-based proteomics we profile the total proteome and secretome of Cryptococcus neoformans wild-type (H99) under in vitro growth conditions. We also define the infectome of C. neoformans and BALB/c macrophages in single runs using high resolution mass spectrometry on a Quadrupole Orbitrap instrument. The in vitro and infectome datasets were integrated using Perseus and candidate fungal proteins of interest were prioritized based on predicted secretory roles, novelty, and abundance profiles. Deletion strains were constructed by double-joint PCR and characterized by phenotypic screening and cell death assays. Virulence-associated candidates will be evaluated in a murine infection model and further characterized by immunofluorescence and interactome analyses. Our preliminary results identify interactions between the host and pathogen critical for disease. Comprehensive profiling of infection from both host and pathogen perspectives unveils new anti-virulence strategies to combat fungal infection.