Identification and characterization of fungal extracellular vesicles in patients with Candidiasis, Cryptococcosis and Paracoccidioidomycosis

Abstract

The incidence and prevalence of fungal infections have increased, and it is important to study the mechanisms that involve the biology of pathogenic fungi to better understand the pathogen-host interaction. Candidiasis, Cryptococcosis and Paracoccidioidomycosis are systemic mycoses caused by fungi of the genera Candida spp., Cryptococcus spp. and Paracoccidoides spp., respectively. Extracellular Vesicles (EVs) consist of spherical lipid bilayers produced by eukaryotes, archaic and bacteria. In pathogenic fungi, EVs can carry virulence factors and other important molecules, promoting the interaction of fungal cells with other organisms, thus playing an important role in pathogenesis and immunobiological interaction. Despite a vast literature related to the production of extracellular vesicles by fungal pathogens, there are still few clinical studies characterizing and showing the role of vesicles in the course of in vivo disease. Thus, we propose to identify fungal extracellular vesicles present in the samples of patients with Candidiasis, Cryptococcosis and Paracoccidioidomycosis. More specifically, (1) purify and characterize the EVs of the clinical samples of patients with fungal infections; (2) identify ergosterol and other secondary metabolites in the EVs obtained; (3) analyze the immunobiological activity of the EVs by incubation with bone marrow-derived macrophages; as well as (4) analyze the induced immune response profile. Our expectation is that the identification of fungal EVs in samples from patients affected with fungal infections, as well as a better understanding of their role in immunoregulation, can help us better understand the impact of EVs in the course of infection as well as its potential in the development of new targets and therapeutic strategies. This project complements our group's collaboration with Johns Hopkins University. (AU)